3,470 results on '"TRPC Cation Channels"'
Search Results
102. TRP channels in health and disease at a glance
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Haoxing Xu and Lixia Yue
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0301 basic medicine ,animal structures ,TRPP Cation Channels ,TRPML ,TRPV Cation Channels ,TRPP ,Biology ,TRPV ,03 medical and health sciences ,Transient receptor potential channel ,0302 clinical medicine ,Transient Receptor Potential Channels ,TRPM ,Ankyrin ,TRPA ,Cell Science at A Glance ,Animals ,Humans ,TRPC ,TRPC Cation Channels ,chemistry.chemical_classification ,Mammals ,Ion Transport ,Cell Biology ,Cell biology ,030104 developmental biology ,chemistry ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
The transient receptor potential (TRP) channel superfamily consists of a large group of non-selective cation channels that serve as cellular sensors for a wide spectrum of physical and environmental stimuli. The 28 mammalian TRPs, categorized into six subfamilies, including TRPC (canonical), TRPV (vanilloid), TRPM (melastatin), TRPA (ankyrin), TRPML (mucolipin) and TRPP (polycystin), are widely expressed in different cells and tissues. TRPs exhibit a variety of unique features that not only distinguish them from other superfamilies of ion channels, but also confer diverse physiological functions. Located at the plasma membrane or in the membranes of intracellular organelles, TRPs are the cellular safeguards that sense various cell stresses and environmental stimuli and translate this information into responses at the organismal level. Loss- or gain-of-function mutations of TRPs cause inherited diseases and pathologies in different physiological systems, whereas up- or down-regulation of TRPs is associated with acquired human disorders. In this Cell Science at a Glance article and the accompanying poster, we briefly summarize the history of the discovery of TRPs, their unique features, recent advances in the understanding of TRP activation mechanisms, the structural basis of TRP Ca2+ selectivity and ligand binding, as well as potential roles in mammalian physiology and pathology.
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- 2021
103. Inhibition of TRPC1, TRPM4 and CHRNA6 ion channels ameliorates depression-like behavior in rats
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İlay Kavuran Buran, Ebru Onalan Etem, and Ahmet Tektemur
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Male ,Behavior, Animal ,Depression ,Brain ,TRPM Cation Channels ,Nicotinic Antagonists ,Receptors, Nicotinic ,Ion Channels ,Rats ,Behavioral Neuroscience ,Disease Models, Animal ,MicroRNAs ,Animals ,Rats, Wistar ,Stress, Psychological ,TRPC Cation Channels - Abstract
The roles of ion channels, miRNAs and, neurotransmitters in the pathophysiology of major depressive disorder (MDD) are not yet fully elucidated. The current study aims to investigate ion channel gene expressions in the brain, the therapeutic efficacies of TRPC1, TRPM4, and CHRNA6 inhibitors, miRNAs specific to these ion channels and, neurotransmitter interactions in a chronic unpredictable mild stress (CUMS) induced MDD rat model. 48 two-month-old male albino Wistar rats were divided into Control, CUMS, Sham, CUMS+Pico145 (TRPC1 inhibitor), CUMS+ 9-Phe (TRPM4 inhibitor), and CUMS+BPiDl (CHRNA6 inhibitor) groups. Seven-week CUMS was used to induce MDD. Inhibitors were administered subacutely on the final of CUMS. Rats were subjected to behavioral tests. Gene expression levels were analyzed using qRT-PCR and neurotransmitter levels using ELISA. CUMS lead to a significant upregulation in the expression of channels in the hippocampus, and channels in the prefrontal cortex. Behavioral experiments determined the antidepressant effects as follows: Pico145 BPiDl 9-Phe. Compared to the Control, serotonin and noradrenaline levels remained unchanged, whereas dopamine levels increased. Acetylcholine levels decreased in CUMS and CUMS+Pico145 groups. CUMS significantly altered the expression of 6 miRNAs in the brain. BPiDl upregulated the expression of miR-6334 and Pico145 upregulated the expression of miR-135b-5p and miR-875 in the prefrontal cortex. The interactions of ion channels, miRNAs, and disruptions of neurotransmitter networks can play an important role in the pathophysiology of MDD. Moreover, as shown in this study, ion channel inhibitors have significant potential in the treatment of this disease.
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- 2021
104. Inhibition of TRPC4/5 Channel Is Effective in Protecting against Histamine-Induced Hyperpermeability by Blocking Ca 2+ Influx in Lung Microvascular Endothelial Cells.
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Zhou Y, Zhang R, and Dong M
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- Humans, Histamine pharmacology, Histamine metabolism, TRPC Cation Channels, Lung, Calcium metabolism, Endothelial Cells metabolism, Transient Receptor Potential Channels metabolism
- Abstract
Dysfunction of lung microvascular endothelium is a major feature in the pathobiology of pulmonary edema and hypoxic respiratory failure. Histamine induces lung microvascular endothelial barrier disruption and hyperpermeability upon evoking intracellular Ca
2+ ([Ca2+ ]i ) dynamics via binding to its receptors. Transient receptor potential canonical (TRPC) channels are Ca2+ -permeable channel and stimulated by the agonists of G-protein-coupled receptors (GPCR). Here, we assessed histamine induced [Ca2+ ]i increases in human lung microvascular endothelial cells (HLMVEC) by using live cell Ca2+ imaging. We found that histamine increased [Ca2+ ]i was maintained at a static elevated level after a transient peak. The elevated Ca2+ plateau was vanished when extracellular Ca2+ was removed, indicating Ca2+ influx from extracellular mediated the histamine-induced Ca2+ plateau. TRPC4/5 channels antagonists, ML204 (10 µM) and HC070 (1 µM), significantly inhibited the Ca2+ plateaus, which was not influenced by Pyr3 or larixyl, the antagonists of TRPC3/6. Furthermore, ML204 or HC070 effectively suppressed the permeability response to histamine in HLMVEC. Our results indicated that histamine-induced Ca2+ influx may be mediated by TRPC4/5 channels and the antagonist of the channel significantly improved histamine-induced HLMVEC dysfunction.- Published
- 2023
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105. Optical Activation of Photoswitchable TRPC Ligands in the Mammalian Olfactory System Using Laser Scanning Confocal Microscopy.
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Ojha NK, Zufall F, and Leinders-Zufall T
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- Animals, Mice, Ligands, Microscopy, Confocal, Mammals, TRPC Cation Channels, Sensory Receptor Cells, B-Lymphocytes
- Abstract
The transient receptor potential canonical (TRPC) ion channels play important biological roles, but their activation mechanisms are incompletely understood. Here, we describe recent methodological advances using small molecular probes designed for photopharmacology of TRPC channels by focusing on results obtained from the mouse olfactory system. These studies developed and used photoswitchable diacylglycerol (DAG) analogs for ultrarapid activation of native TRPC2 channels in vomeronasal sensory neurons and type B cells of the main olfactory epithelium. Further studies investigated the role of TRPC5 channels in prolactin regulation of dopamine neurons in the arcuate nucleus of the hypothalamus. Here, the first photoswitchable TRPC5 modulator, BTDAzo, was developed and shown to control endogenous TRPC5-based neuronal Ca
2+ responses in mouse brain slices. Thus, photoswitchable reagents are rapidly gaining widespread recognition for investigating various types of TRPC channels including TRPC2, TRPC3, TRPC5, and TRPC6, enabling to gain new insights into the gating mechanisms and functions of these channels., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2023
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106. TRPC1 channels regulate the activation of pancreatic stellate cells through ERK1/2 and SMAD2 pathways and perpetuate their pressure-mediated activation
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Silviya, Radoslavova, Benedikt, Fels, Zoltan, Pethö, Matthias, Gruner, Tobias, Ruck, Sven G, Meuth, Antoine, Folcher, Natalia, Prevarskaya, Albrecht, Schwab, Halima, Ouadid-Ahidouch, Laboratoire de Physiologie Cellulaire et Moléculaire - UR UPJV 4667 (LPCM), Université de Picardie Jules Verne (UPJV), Rôle des canaux ioniques membranaires et du calcium intracellulaire dans la physiopathologie de la prostate, Université de Lille, Sciences et Technologies-Institut National de la Santé et de la Recherche Médicale (INSERM), University Hospital Düsseldorf, Laboratoire de Physiologie Cellulaire : Canaux ioniques, inflammation et cancer - U 1003 (PHYCELL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, and Westfälische Wilhelms-Universität Münster = University of Münster (WWU)
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Pancreatic stellate cells ,High-pressure ,MAP Kinase Signaling System ,Physiology ,Activation ,Smad2 Protein ,Cell Biology ,Pancreatic Neoplasms ,Pancreatic ductal adenocarcinoma ,Mice ,TRPC1 channels ,Tumor Microenvironment ,Animals ,Humans ,Extracellular Signal-Regulated MAP Kinases ,Molecular Biology ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology ,Carcinoma, Pancreatic Ductal ,TRPC Cation Channels - Abstract
International audience; Pancreatic stellate cell (PSC) activation is a major event occurring during pancreatic ductal adenocarcinoma (PDAC) development. Up to now mechanisms underlying their activation by mechanical cues such as the elevated tissue pressure in PDAC remain poorly understood. Here we investigate the role of one potential mechano-transducer, TRPC1 ion channel, in PSC activation. Using pre-activated human siTRPC1 and murine TRPC1-KO PSCs, we show that TRPC1 promotes αSMA (α-smooth muscle actin) expression, the main activation marker, in cooperation with the phosphorylated SMAD2, under normal and elevated pressure. Functional studies following TRPC1 silencing demonstrate the dual role of TRPC1 in the modulation of PSC proliferation and IL-6 secretion through the activation of ERK1/2 and SMAD2 pathways. Moreover, pressurization changes the mechanical behavior of PSCs by increasing their cellular stiffness and emitted traction forces in a TRPC1-dependent manner. In summary, these results point to a role of TRPC1 channels in sensing and transducing the characteristic mechanical properties of the PDAC microenvironment in PSCs.
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- 2022
107. Sertoli cell transplantation attenuates microglial activation and inhibits TRPC6 expression in neuropathic pain induced by spinal cord injury
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Behnaz, Rahimi, Abbas, Aliaghaei, Fatemeh, Ramezani, Zahra, Behroozi, and Farinaz, Nasirinezhad
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Male ,Analgesics ,Sertoli Cells ,Cell Transplantation ,TOR Serine-Threonine Kinases ,Experimental and Cognitive Psychology ,Rats ,Rats, Sprague-Dawley ,Behavioral Neuroscience ,Spinal Cord ,Hyperalgesia ,TRPC6 Cation Channel ,Animals ,Neuralgia ,Microglia ,Rats, Wistar ,Spinal Cord Injuries ,TRPC Cation Channels - Abstract
Cell therapy is a promising treatment method for relieving neuropathic pain caused by spinal cord injuries (SCI). Sertoli cells (SCs) are an attractive choice given their demonstrated secretion of growth factors and immunosuppressant effect. This study mechanistically characterizes the analgesic effect of SCs transplantation.The clip compression SCI model was carried out on the T12-T13 level in male Wistar rats. One-week post-SCI, SCs were transplanted into the site of injury. Animals underwent Basso, Beattie, and Bresnahan locomotor scoring, mechanical allodynia, and thermal hyperalgesia on a weekly basis for a duration of six weeks. Histological examination of the spinal cord and molecular evaluation of Iba-1, P2Y4, TRPC6, and P-mTOR were performed. SCs survival, measured by anti-Müllerian hormone expression in the spinal cord.Animals that received SCs transplantation showed improvement in motor function recovery and pain relief. Furthermore, a cavity was significantly decreased in the transplanted animals (p = 0.0024), the expression level of TRPC6 and caspase3 and the number of activated microglia decreased compared to the SCI animals, and p-mTOR and P2Y4R expression remarkably increased compared to the SCI group.SCs transplantation produces an analgesic effect which may represent a promising treatment for SCI-induced chronic pain.
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- 2022
108. TRPC1 contributes to endotoxemia-induced myocardial dysfunction via mediating myocardial apoptosis and autophagy
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Wen, Tian, Shao-Yuan, Liu, Meng, Zhang, Jing-Ru, Meng, Na, Tang, Ying-Da, Feng, Yang, Sun, Yuan-Yuan, Gao, Lei, Zhou, Wei, Cao, and Xiao-Qiang, Li
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Lipopolysaccharides ,Mice, Inbred C57BL ,Pharmacology ,Mice ,Heart Diseases ,Calpain ,Caveolin 1 ,Autophagy ,Animals ,Apoptosis ,Myocytes, Cardiac ,Endotoxemia ,TRPC Cation Channels - Abstract
Cardiac dysfunction is a vital complication of endotoxemia (ETM) with limited therapeutic options. Transient receptor potential canonical channel (TRPC)1 was involved in various heart diseases. While, the role of TRPC1 in ETM-induced cardiac dysfunction remains to be defined. In this study, we found that TRPC1 protein expression was significantly upregulated in hearts of lipopolysaccharide (LPS)-challenged mice. What's more, TRPC1 knockdown significantly alleviated LPS-induced cardiac dysfunction and injury. Further myocardial mRNA-sequencing analysis revealed that TRPC1 might participate in pathogenesis of ETM-induced cardiac dysfunction via mediating myocardial apoptosis and autophagy. Data showed that knockdown of TRPC1 significantly ameliorated LPS-induced myocardial apoptotic injury, cardiomyocytes autophagosome accumulation, and myocardial autophagic flux. Simultaneously, deletion of TRPC1 reversed LPS-induced molecular changes of apoptosis/autophagy signaling pathway in cardiomyocytes. Moreover, TRPC1 could promote LPS-triggered intracellular Ca
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- 2022
109. Transient Receptor Potential C 1/4/5 Is a Determinant of MTI-101 Induced Calcium Influx and Cell Death in Multiple Myeloma
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Brandon E. Johnson, Lori A. Hazlehurst, Wei-Chih Chen, Gangqing Hu, Osama M Elzamzamy, and Reinhold Penner
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SERCA ,TRPC ,QH301-705.5 ,TRPC5 ,Peptides, Cyclic ,Article ,TRPC1 ,Transient receptor potential channel ,Cell Line, Tumor ,Extracellular ,Humans ,Calcium Signaling ,Biology (General) ,TRPC Cation Channels ,calcium ,Cell Death ,biology ,Chemistry ,STIM1 ,Calpain ,General Medicine ,Neoplasm Proteins ,multiple myeloma ,Protein Transport ,Cancer research ,biology.protein ,human activities ,SOCE - Abstract
Multiple myeloma (MM) is a currently incurable hematologic cancer. Patients that initially respond to therapeutic intervention eventually relapse with drug resistant disease. Thus, novel treatment strategies are critically needed to improve patient outcomes. Our group has developed a novel cyclic peptide referred to as MTI-101 for the treatment of MM. We previously reported that acquired resistance to HYD-1, the linear form of MTI-101, correlated with the repression of genes involved in store operated Ca2+ entry (SOCE): PLCβ, SERCA, ITPR3, and TRPC1 expression. In this study, we sought to determine the role of TRPC1 heteromers in mediating MTI-101 induced cationic flux. Our data indicate that, consistent with the activation of TRPC heteromers, MTI-101 treatment induced Ca2+ and Na+ influx. However, replacing extracellular Na+ with NMDG did not reduce MTI-101-induced cell death. In contrast, decreasing extracellular Ca2+ reduced both MTI-101-induced Ca2+ influx as well as cell death. The causative role of TRPC heteromers was established by suppressing STIM1, TRPC1, TRPC4, or TRPC5 function both pharmacologically and by siRNA, resulting in a reduction in MTI-101-induced Ca2+ influx. Mechanistically, MTI-101 treatment induces trafficking of TRPC1 to the membrane and co-immunoprecipitation studies indicate that MTI-101 treatment induces a TRPC1-STIM1 complex. Moreover, treatment with calpeptin inhibited MTI-101-induced Ca2+ influx and cell death, indicating a role of calpain in the mechanism of MTI-101-induced cytotoxicity. Finally, components of the SOCE pathway were found to be poor prognostic indicators among MM patients, suggesting that this pathway is attractive for the treatment of MM.
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- 2021
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110. (-)-Englerin-A Has Analgesic and Anti-Inflammatory Effects Independent of TRPC4 and 5
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Joao de Sousa Valente, Susan D. Brain, Khadija M. Alawi, Sabah Bharde, Ali A Zarban, Xenia Kodji, Brentton Barrett, István Nagy, Fulye Argunhan, and Dibesh Thapa
- Subjects
0301 basic medicine ,Male ,Chemistry, Multidisciplinary ,Anti-Inflammatory Agents ,Pharmacology ,Carrageenan ,TRPC4 ,TRPC5 ,chemistry.chemical_compound ,Sesquiterpenes, Guaiane ,0302 clinical medicine ,CHANNEL ,Dorsal root ganglion ,Ganglia, Spinal ,Edema ,Biology (General) ,NEURONS ,Spectroscopy ,Cells, Cultured ,Mice, Knockout ,Analgesics ,Behavior, Animal ,General Medicine ,Cobalt ,Computer Science Applications ,Chemistry ,medicine.anatomical_structure ,Phenotype ,Hyperalgesia ,Physical Sciences ,ROOT GANGLION-CELLS ,medicine.symptom ,Life Sciences & Biomedicine ,Agonist ,Biochemistry & Molecular Biology ,ENZYME ,Sensory Receptor Cells ,medicine.drug_class ,QH301-705.5 ,0699 Other Biological Sciences ,Analgesic ,Pain ,Inflammation ,Article ,Catalysis ,Anti-inflammatory ,Inorganic Chemistry ,03 medical and health sciences ,In vivo ,ENGLERIN ,0399 Other Chemical Sciences ,medicine ,Animals ,Physical and Theoretical Chemistry ,Molecular Biology ,QD1-999 ,TRPC Cation Channels ,0604 Genetics ,Science & Technology ,Chemical Physics ,IDENTIFICATION ,POTENT ,Organic Chemistry ,Antagonist ,Disease Models, Animal ,030104 developmental biology ,chemistry ,(-)-Englerin A ,RAT ,synovitis ,030217 neurology & neurosurgery - Abstract
Recently, we found that the deletion of TRPC5 leads to increased inflammation and pain-related behaviour in two animal models of arthritis. (-)-Englerin A (EA), an extract from the East African plant Phyllanthus engleri has been identified as a TRPC4/5 agonist. Here, we studied whether or not EA has any anti-inflammatory and analgesic properties via TRPC4/5 in the carrageenan model of inflammation. We found that EA treatment in CD1 mice inhibited thermal hyperalgesia and mechanical allodynia in a dose-dependent manner. Furthermore, EA significantly reduced the volume of carrageenan-induced paw oedema and the mass of the treated paws. Additionally, in dorsal root ganglion (DRG) neurons cultured from WT 129S1/SvIm mice, EA induced a dose-dependent cobalt uptake that was surprisingly preserved in cultured DRG neurons from 129S1/SvIm TRPC5 KO mice. Likewise, EA-induced anti-inflammatory and analgesic effects were preserved in the carrageenan model in animals lacking TRPC5 expression or in mice treated with TRPC4/5 antagonist ML204.This study demonstrates that while EA activates a sub-population of DRG neurons, it induces a novel TRPC4/5-independent analgesic and anti-inflammatory effect in vivo. Future studies are needed to elucidate the molecular and cellular mechanisms underlying EA’s anti-inflammatory and analgesic effects.
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- 2021
111. Increased transient receptor potential canonical 3 activity is involved in the pathogenesis of detrusor overactivity by dynamic interaction with Na
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Jingzhen, Zhu, Yi, Fan, Qudong, Lu, Yang, Yang, Hui, Li, Xin, Liu, Hengshuai, Zhang, Bishao, Sun, Qian, Liu, Jiang, Zhao, Zhenxing, Yang, Longkun, Li, Huan, Feng, and Jie, Xu
- Subjects
Aniline Compounds ,Reverse Transcriptase Polymerase Chain Reaction ,Urinary Bladder, Overactive ,Phenyl Ethers ,Myocytes, Smooth Muscle ,Urinary Bladder ,Sodium-Calcium Exchanger ,Rats ,Urinary Bladder Neck Obstruction ,Disease Models, Animal ,Gene Expression Regulation ,Animals ,Humans ,Calcium ,Female ,Cells, Cultured ,Protein Binding ,TRPC Cation Channels - Abstract
Transient receptor potential canonical 3 (TRPC3) is a nonselective cation channel, and its dysfunction is the basis of many clinical diseases. However, little is known about its possible role in the bladder. The purpose of this study was to explore the function and mechanism of TRPC3 in partial bladder outlet obstruction (PBOO)-induced detrusor overactivity (DO). We studied 31 adult female rats with DO induced by PBOO (the DO group) and 40 sham-operated rats (the control group). Here we report that the expression of TRPC3 in the bladder of DO rats increased significantly. Furthermore, PYR10, which can selectively inhibit the TRPC3 channel, significantly reduced bladder excitability in DO and control rats, but the decrease of the bladder excitability of DO rats was more obvious. PYR10 significantly reduced the intracellular calcium concentration in smooth muscle cells (SMCs) in DO and control rats. Finally, Na
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- 2021
112. Structural mechanism of human TRPC3 and TRPC6 channel regulation by their intracellular calcium-binding sites
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Wenjun Guo, Qinglin Tang, Miao Wei, Yunlu Kang, Jing-Xiang Wu, and Lei Chen
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Binding Sites ,Glomerulosclerosis, Focal Segmental ,General Neuroscience ,TRPC6 Cation Channel ,Humans ,Calcium ,Calcium Channels ,TRPC Cation Channels - Abstract
TRPC3 and TRPC6 channels are calcium-permeable non-selective cation channels that are involved in many physiological processes. The gain-of-function (GOF) mutations of TRPC6 lead to familial focal segmental glomerulosclerosis (FSGS) in humans, but their pathogenic mechanism remains elusive. Here, we report the cryo-EM structures of human TRPC3 in both high-calcium and low-calcium conditions. Based on these structures and accompanying electrophysiological studies, we identified both inhibitory and activating calcium-binding sites in TRPC3 that couple intracellular calcium concentrations to the basal channel activity. These calcium sensors are also structurally and functionally conserved in TRPC6. We uncovered that the GOF mutations of TRPC6 activate the channel by allosterically abolishing the inhibitory effects of intracellular calcium. Furthermore, structures of human TRPC6 in complex with two chemically distinct inhibitors bound at different ligand-binding pockets reveal different conformations of the transmembrane domain, providing templates for further structure-based drug design targeting TRPC6-related diseases such as FSGS.
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- 2021
113. TRPC1‐mediated Ca2+ signaling enhances intestinal epithelial restitution by increasing α4 association with PP2Ac after wounding
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Hee Kyoung Chung, Navneeta Rathor, Jaladanki N. Rao, Jian-Ying Wang, Shelley R. Wang, and Jia-Le Song
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cell migration ,epithelial restitution ,TRPC1 ,Physiology ,Protein subunit ,polyamines ,mucosal injury ,030204 cardiovascular system & hematology ,IEC‐6 cells ,Ornithine decarboxylase ,Cell Line ,03 medical and health sciences ,Transient receptor potential channel ,0302 clinical medicine ,Cell Movement ,Physiology (medical) ,QP1-981 ,Gene silencing ,ornithine decarboxylase ,Animals ,Calcium Signaling ,Protein Phosphatase 2 ,Adaptor Proteins, Signal Transducing ,TRPC Cation Channels ,Wound Healing ,Chemistry ,Cell migration ,Protein phosphatase 2 ,Transfection ,Original Articles ,Cell biology ,Rats ,Enterocytes ,Original Article ,030217 neurology & neurosurgery ,Molecular Chaperones - Abstract
Gut epithelial restitution after superficial wounding is an important repair modality regulated by numerous factors including Ca2+ signaling and cellular polyamines. Transient receptor potential canonical‐1 (TRPC1) functions as a store‐operated Ca2+ channel in intestinal epithelial cells (IECs) and its activation increases epithelial restitution by inducing Ca2+ influx after acute injury. α4 is a multiple functional protein and implicated in many aspects of cell functions by modulating protein phosphatase 2A (PP2A) stability and activity. Here we show that the clonal populations of IECs stably expressing TRPC1 (IEC‐TRPC1) exhibited increased levels of α4 and PP2A catalytic subunit (PP2Ac) and that TRPC1 promoted intestinal epithelial restitution by increasing α4/PP2Ac association. The levels of α4 and PP2Ac proteins increased significantly in stable IEC‐TRPC1 cells and this induction in α4/PP2Ac complexes was accompanied by an increase in IEC migration after wounding. α4 silencing by transfection with siRNA targeting α4 (siα4) or PP2Ac silencing destabilized α4/PP2Ac complexes in stable IEC‐TRPC1 cells and repressed cell migration over the wounded area. Increasing the levels of cellular polyamines by stable transfection with the Odc gene stimulated α4 and PP2Ac expression and enhanced their association, thus also promoting epithelial restitution after wounding. In contrast, depletion of cellular polyamines by treatment with α‐difluoromethylornithine reduced α4/PP2Ac complexes and repressed cell migration. Ectopic overexpression of α4 partially rescued rapid epithelial repair in polyamine‐deficient cells. These results indicate that activation of TRPC1‐mediated Ca2+ signaling enhances cell migration primarily by increasing α4/PP2Ac associations after wounding and this pathway is tightly regulated by cellular polyamines.
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- 2021
114. Chronic hypoxia promoted pulmonary arterial smooth muscle cells proliferation through upregulated calcium-sensing receptorcanonical transient receptor potential 1/6 pathway
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Weitao Cao, Yongliang Jiang, Lingmei Huang, Xing Wen, Zhenli Fu, Shaoxing Li, Rongmin Liu, Gongyong Peng, Pixin Ran, Wei Hong, Bing Li, Juan Xu, and Jinding Pu
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Physiology ,Hypertension, Pulmonary ,Myocytes, Smooth Muscle ,030204 cardiovascular system & hematology ,Pulmonary Artery ,Muscle, Smooth, Vascular ,TRPC6 ,TRPC1 ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Physiology (medical) ,TRPC6 Cation Channel ,Animals ,Viability assay ,Receptor ,Hypoxia ,Molecular Biology ,TRPC ,Cells, Cultured ,Cell Proliferation ,TRPC Cation Channels ,DNA synthesis ,Chemistry ,Cell growth ,DNA ,Cell biology ,Rats ,Calcium ,Cardiology and Cardiovascular Medicine ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Objectives Although both calcium-sensing receptor (CaSR) and canonical transient receptor potential (TRPC) proteins contribute to chronic hypoxia (CH)-induced pulmonary arterial smooth muscle cells (PASMCs) proliferation, the relationship between CaSR and TRPC in hypoxic PASMCs proliferation remains poorly understood. The goal of this study was to identify that CH promotes PASMCs proliferation through CaSR-TRPC pathway. Methods Rat PASMCs were isolated and treated with CH. Cell proliferation was assessed by cell counting, CCK-8 assay, and EdU incorporation. CaSR and TRPC expressions were determined by qPCR and Western blotting. Store-operated Ca2+ entry (SOCE) was assessed by extracellular Ca2+ restoration. Results In PASMCs, CH enhanced the cell number, cell viability and DNA synthesis, which is accompanied by upregulated expression of CaSR, TRPC1 and TRPC6. Negative CaSR modulators (NPS2143, NPS2390) inhibited, whereas positive modulators (spermine, R568) enhanced, the CH-induced increases in cell number, cell viability and DNA synthesis in PASMCs. Knockdown of CaSR by siRNA inhibited the CH-induced upregulation of TRPC1 and TRPC6 and enhancement of SOCE and attenuated the CH-induced enhancements of cell number, cell viability and DNA synthesis in PASMCs. However, neither siTRPC1 nor siTRPC6 had an effect on the CH-induced CaSR upregulation, although both significantly attenuated the CH-induced enhancements of cell number, cell viability and DNA synthesis in PASMCs. Conclusion These results demonstrate that upregulated CaSR-TRPC1/6 pathway mediating PASMCs proliferation is an important pathogenic mechanism under hypoxic conditions.
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- 2021
115. Correlation between classical transient receptor potential channel 1 gene polymorphism and microalbuminuria in patients with primary hypertension
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Xinjuan Xu, Yaping Cao, Yulan Chen, Yu Li, Yu Zhang, and Bumairemu Maitikuerban
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Male ,medicine.medical_specialty ,Physiology ,Single-nucleotide polymorphism ,030204 cardiovascular system & hematology ,Polymorphism, Single Nucleotide ,Correlation ,TRPC1 ,03 medical and health sciences ,Transient receptor potential channel ,0302 clinical medicine ,Gene Frequency ,Risk Factors ,Internal medicine ,Internal Medicine ,medicine ,Albuminuria ,Humans ,In patient ,030212 general & internal medicine ,TRPC Cation Channels ,business.industry ,General Medicine ,Middle Aged ,medicine.disease ,Endocrinology ,Logistic Models ,Microalbuminuria ,Female ,Gene polymorphism ,Essential Hypertension ,business - Abstract
To investigate the correlation between transient receptor potential channel 1 ((1) No significant differences were detected in gender, smoking history, drinking history, family history, course of hypertension, fasting blood glucose, urea, creatinine, triglyceride, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, glycosylated hemoglobin, vitamin D, homocysteine, and cystatin C between the two groups (
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- 2021
116. Electrophysiological Properties of Endogenous Single Ca
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Dmitrii, Kolesnikov, Anastasiia, Perevoznikova, Konstantin, Gusev, Lyubov, Glushankova, Elena, Kaznacheyeva, and Alexey, Shalygin
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Patch-Clamp Techniques ,single channel ,TRPC1 ,Cations, Divalent ,TMEM16 ,HEK293 ,Anoctamins ,anoctamin ,Article ,HEK293 Cells ,Chloride Channels ,CaCC ,Humans ,Calcium ,Phospholipid Transfer Proteins ,IP3R ,TRPC Cation Channels ,SOCE - Abstract
Microdomains formed by proteins of endoplasmic reticulum and plasma membrane play a key role in store-operated Ca2+ entry (SOCE). Ca2+ release through inositol 1,4,5-trisphosphate receptor (IP3R) and subsequent Ca2+ store depletion activate STIM (stromal interaction molecules) proteins, sensors of intraluminal Ca2+, which, in turn, open the Orai channels in plasma membrane. Downstream to this process could be activated TRPC (transient receptor potential-canonical) calcium permeable channels. Using single channel patch-clamp technique we found that a local Ca2+ entry through TRPC1 channels activated endogenous Ca2+-activated chloride channels (CaCCs) with properties similar to Anoctamin6 (TMEM16F). Our data suggest that their outward rectification is based on the dependence from membrane potential of both the channel conductance and the channel activity: (1) The conductance of active CaCCs highly depends on the transmembrane potential (from 3 pS at negative potentials till 60 pS at positive potentials); (2) their activity (NPo) is enhanced with increasing Ca2+ concentration and/or transmembrane potential, conversely lowering of intracellular Ca2+ concentration reduced the open state dwell time; (3) CaCC amplitude is only slightly increased by intracellular Ca2+ concentration. Experiments with Ca2+ buffering by EGTA or BAPTA suggest close local arrangement of functional CaCCs and TRPC1 channels. It is supposed that Ca2+-activated chloride channels are involved in Ca2+ entry microdomains.
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- 2021
117. Calmodulin binds to Drosophila TRP with an unexpected mode
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Craig Montell, Weidi Chen, Mingjie Zhang, Ziling Sun, Fei Ye, Sabrina Asteriti, Wei Liu, Jun Wan, Roger C. Hardie, Zijing Chen, Zeyu Shen, Hardie, Roger [0000-0001-5531-3264], and Apollo - University of Cambridge Repository
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calmodulin ,visual signal transduction ,animal structures ,Calmodulin ,TRPC4 ,03 medical and health sciences ,Transient receptor potential channel ,Mice ,Transient Receptor Potential Channels ,Structural Biology ,Negative feedback ,Animals ,Drosophila Proteins ,Humans ,Molecular Biology ,Drosophila TRP ,030304 developmental biology ,TRPC Cation Channels ,0303 health sciences ,Binding Sites ,biology ,Chemistry ,030302 biochemistry & molecular biology ,Visual Signal Transduction ,Ca(2+)-dependent target binding ,Drosophila melanogaster ,HEK293 Cells ,Mutation ,biology.protein ,Molecular mechanism ,Biophysics ,Calcium ,Linker ,Visual phototransduction ,Protein Binding - Abstract
Drosophila TRP is a calcium-permeable cation channel essential for fly visual signal transduction. During phototransduction, Ca2+ mediates both positive and negative feedback regulation on TRP channel activity, possibly via binding to calmodulin (CaM). However, the molecular mechanism underlying Ca2+ modulated CaM/TRP interaction is poorly understood. Here, we discover an unexpected, Ca2+-dependent binding mode between CaM and TRP. The TRP tail contains two CaM binding sites (CBS1 and CBS2) separated by an ∼70-residue linker. CBS1 binds to the CaM N-lobe and CBS2 recognizes the CaM C-lobe. Structural studies reveal the lobe-specific binding of CaM to CBS1&2. Mutations introduced in both CBS1 and CBS2 eliminated CaM binding in full-length TRP, but surprisingly had no effect on the response to light under physiological conditions, suggesting alternative mechanisms governing Ca2+-mediated feedback on the channel activity. Finally, we discover that TRPC4, the closest mammalian paralog of Drosophila TRP, adopts a similar CaM binding mode.
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- 2021
118. PIP2 regulation of TRPC5 channel activation and desensitization
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Anna Greka, Leigh D. Plant, Mehek Ningoo, and Diomedes E. Logothetis
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0301 basic medicine ,TRPC5 channels ,Models, Molecular ,Phosphatidylinositol 4,5-Diphosphate ,transient receptor potential channels (TRP channels) ,CCh, carbachol ,CIBN, cryptochrome-interacting basic-helix-loop-helix N-terminal fragment ,Protein Conformation ,NHERF, Na+/H+ exchanger regulatory factor ,Gating ,CRY2, cryptochrome 2 ,TRPC5 ,Biochemistry ,OAG, 1-oleoyl-2-acetyl-sn-glycerol ,TIRF, total internal reflection fluorescence ,03 medical and health sciences ,Transient receptor potential channel ,chemistry.chemical_compound ,PLC, phospholipase C ,Humans ,Molecular Biology ,Protein kinase C ,Ion channel ,Diacylglycerol kinase ,diC8–PIP2, dioctanoyl-glycerol-PIP2 ,TRPC Cation Channels ,030102 biochemistry & molecular biology ,Phospholipase C ,Chemistry ,PIP-5K, phosphatidylinositol 4-phosphate 5 kinase ,phosphatidylinositol 4,5-bisphosphate (PIP2) ,Cell Biology ,Editors' Pick ,diacyl glycerol (DAG) ,TRPC5, transient receptor potential canonical type 5 ,YFP, yellow fluorescent protein ,Cell biology ,phosphoinositide ,PIP2, phosphatidylinositol 4,5-bisphosphate ,PMA, phorbol 12-myristate-13-acetate ,030104 developmental biology ,HEK293 Cells ,Phosphatidylinositol 4,5-bisphosphate ,lipids (amino acids, peptides, and proteins) ,phosphatidylinositol signaling ,IP3, inositol 1,4,5-triphosphate ,5’-ptaseOCRL, 5’-phosphatase domain of OCRL ,DAG, diacylglycerol ,Research Article - Abstract
Transient receptor potential canonical type 5 (TRPC5) ion channels are expressed in the brain and kidney and have been identified as promising therapeutic targets whose selective inhibition can protect against diseases driven by a leaky kidney filter, such as focal segmental glomerular sclerosis. TRPC5 channels are activated not only by elevated levels of extracellular Ca2+or lanthanide ions but also by G protein (Gq/11) stimulation. Phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis by phospholipase C enzymes leads to PKC-mediated phosphorylation of TRPC5 channels and their subsequent desensitization. However, the roles of PIP2 in activation and maintenance of TRPC5 channel activity via its hydrolysis product diacyl glycerol (DAG), as well as the mechanism of desensitization of TRPC5 activity by DAG-stimulated PKC activity, remain unclear. Here, we designed experiments to distinguish between the processes underlying channel activation and inhibition. Employing whole-cell patch-clamp, we used an optogenetic tool to dephosphorylate PIP2 and assess channel–PIP2 interactions influenced by activators, such as DAG, or inhibitors, such as PKC phosphorylation. Using total internal reflection microscopy, we assessed channel cell surface density. We show that PIP2 controls both the PKC-mediated inhibition and the DAG- and lanthanide-mediated activation of TRPC5 currents via control of gating rather than channel cell surface density. These mechanistic insights promise to aid in the development of more selective and precise inhibitors to block TRPC5 channel activity and illuminate new opportunities for targeted therapies for a group of chronic kidney diseases for which there is currently a great unmet need.
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- 2021
119. Myocardial Hypertrophy and Fibrosis Are Associated with Cardiomyocyte Beta-Catenin and TRPC6/Calcineurin/NFAT Signaling in Spontaneously Hypertensive Rats with 5/6 Nephrectomy
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Marina Parastaeva, Olga Beresneva, Olga Galkina, Evdokia Bogdanova, Vladimir Dobronravov, Natalia Semenova, Galina Ivanova, and Irina Zubina
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Male ,0301 basic medicine ,calcineurin B ,calcineurin A ,TRPC6 ,030204 cardiovascular system & hematology ,urologic and male genital diseases ,Nephrectomy ,Rats, Inbred WKY ,0302 clinical medicine ,Fibrosis ,Rats, Inbred SHR ,Myocytes, Cardiac ,Biology (General) ,Wnt Signaling Pathway ,beta Catenin ,Spectroscopy ,Kidney ,Ventricular Remodeling ,Calcineurin ,Wnt signaling pathway ,NFAT ,General Medicine ,Computer Science Applications ,Chemistry ,medicine.anatomical_structure ,Hypertension ,cardiovascular system ,medicine.medical_specialty ,arterial hypertension ,QH301-705.5 ,Cardiomegaly ,Article ,Klotho ,Catalysis ,fibroblast growth factor 23 ,Inorganic Chemistry ,03 medical and health sciences ,Internal medicine ,medicine ,Animals ,parathyroid hormone ,cardiovascular diseases ,Renal Insufficiency, Chronic ,Physical and Theoretical Chemistry ,Molecular Biology ,QD1-999 ,TRPC Cation Channels ,NFATC Transcription Factors ,business.industry ,Organic Chemistry ,β-catenin ,medicine.disease ,Fibroblast Growth Factor-23 ,030104 developmental biology ,Endocrinology ,Blood pressure ,Myocardial fibrosis ,cardiac remodeling ,business ,chronic kidney disease ,Kidney disease - Abstract
Background: Arterial hypertension (AH) is associated with heart and chronic kidney disease (CKD). However, the precise mechanisms of myocardial remodeling (MR) in the settings of CKD remain elusive. We hypothesized that TRPC6, calcineurin/NFAT, and Wnt/β-catenin signaling pathways are involved in the development of MR in the background of CKD and AH. Methods: Early CKD was induced by performing a 5/6 nephrectomy (5/6NE) in spontaneously hypertensive rats (SHR-NE). Sham-operated (SO) SHR (SHR-SO) and Wistar Kyoto (WKY-SO) rats served as controls. Systolic blood pressure (SBP), heart rate, myocardial mass index (MMI), serum creatinine, cardiomyocyte diameter (dCM), myocardial fibrosis (MF), serum and kidney α-Klotho levels, myocardial expression of calcineurin (CaN), TRPC6, and β-catenin were measured two months after 5/6NE or SO. Results: NE-induced kidney dysfunction corresponded to mild-to-moderate human CKD and was associated with an increase in FGF23 and a decrease in renal α-Klotho. The levels of SBP, MMI, dCM, and MF were higher in SHRs compared to WKY-SO as well as in SHR-NE vs. SHR-SO. The MR was associated with increased cardiomyocyte expression of CaN/NFAT and β-catenin along with its intracellular re-distribution. TRPC6 protein levels were substantially elevated in both SHR groups with higher Trpc6 mRNA expression in SHR-NE. Conclusions: The Wnt/β-catenin and TRPC6/CaN/NFAT hypertrophic signaling pathways seem to be involved in myocardial remodeling in the settings of AH and CKD and might be mediated by FGF23 and α-Klotho axis.
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- 2021
120. Endoplasmic Reticulum‐Plasma Membrane Contact Sites as an Organizing Principle for Compartmentalized Calcium and cAMP Signaling
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Tim Crul and József Maléth
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Cell signaling ,Orai1 ,QH301-705.5 ,STIM1 ,membrane contact sites ,Review ,Endoplasmic Reticulum ,Models, Biological ,Second Messenger Systems ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Membrane Microdomains ,Spatio-Temporal Analysis ,Stromal Interaction Molecules ,Caveolae ,cAMP ,Cyclic AMP ,Animals ,Humans ,Cyclic adenosine monophosphate ,Calcium Signaling ,Biology (General) ,Physical and Theoretical Chemistry ,QD1-999 ,Molecular Biology ,Lipid raft ,Spectroscopy ,030304 developmental biology ,TRPC Cation Channels ,0303 health sciences ,calcium ,Binding Sites ,ORAI1 ,Endoplasmic reticulum ,Organic Chemistry ,Cell Membrane ,General Medicine ,Calcium Release Activated Calcium Channels ,Computer Science Applications ,Cell biology ,Chemistry ,chemistry ,Second messenger system ,030217 neurology & neurosurgery ,SOCE - Abstract
In eukaryotic cells, ultimate specificity in activation and action—for example, by means of second messengers—of the myriad of signaling cascades is primordial. In fact, versatile and ubiquitous second messengers, such as calcium (Ca2+) and cyclic adenosine monophosphate (cAMP), regulate multiple—sometimes opposite—cellular functions in a specific spatiotemporal manner. Cells achieve this through segregation of the initiators and modulators to specific plasma membrane (PM) subdomains, such as lipid rafts and caveolae, as well as by dynamic close contacts between the endoplasmic reticulum (ER) membrane and other intracellular organelles, including the PM. Especially, these membrane contact sites (MCSs) are currently receiving a lot of attention as their large influence on cell signaling regulation and cell physiology is increasingly appreciated. Depletion of ER Ca2+ stores activates ER membrane STIM proteins, which activate PM-residing Orai and TRPC Ca2+ channels at ER–PM contact sites. Within the MCS, Ca2+ fluxes relay to cAMP signaling through highly interconnected networks. However, the precise mechanisms of MCS formation and the influence of their dynamic lipid environment on their functional maintenance are not completely understood. The current review aims to provide an overview of our current understanding and to identify open questions of the field.
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- 2021
121. Potent, selective, and subunit‐dependent activation of TRPC5 channels by a xanthine derivative
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Katsuhiko Muraki, Aisling Minard, David J. Wright, Isabelle B. Pickles, David J. Beech, Robin S. Bon, Claudia C. Bauer, Melanie J. Ludlow, Stuart L. Warriner, Matthew P. Burnham, and Eulashini Chuntharpursat-Bon
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0301 basic medicine ,Patch-Clamp Techniques ,Cell Survival ,TRPC5 ,Heterocyclic Compounds, 2-Ring ,TRPC4 ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Transient receptor potential channel ,0302 clinical medicine ,TRPC3 ,Humans ,TRPM2 ,Patch clamp ,Cells, Cultured ,TRPC Cation Channels ,Pharmacology ,Dose-Response Relationship, Drug ,Molecular Structure ,Voltage-dependent calcium channel ,Xanthine ,Research Papers ,HEK293 Cells ,030104 developmental biology ,chemistry ,Purines ,Biophysics ,Calcium ,030217 neurology & neurosurgery ,Research Paper - Abstract
Background and purpose The TRPC1, TRPC4, and TRPC5 proteins form homotetrameric or heterotetrameric, calcium-permeable cation channels that are involved in various disease states. Recent research has yielded specific and potent xanthine-based TRPC1/4/5 inhibitors. Here, we investigated the possibility of xanthine-based activators of these channels. Experimental approach An analogue of the TRPC1/4/5 inhibitor Pico145, AM237, was synthesized and its activity was investigated using HEK cells overexpressing TRPC4, TRPC5, TRPC4-C1, TRPC5-C1, TRPC1:C4 or TRPC1:C5 channels, and in A498 cells expressing native TRPC1:C4 channels. TRPC1/4/5 channel activities were assayed by measuring intracellular concentration of Ca2+ ([Ca2+ ]i ) and by patch-clamp electrophysiology. Selectivity of AM237 was tested against TRPC3, TRPC6, TRPV4, or TRPM2 channels. Key results AM237 potently activated TRPC5:C5 channels (EC50 15-20 nM in [Ca2+ ]i assay) and potentiated their activation by sphingosine-1-phosphate but suppressed activation evoked by (-)-englerin A (EA). In patch-clamp studies, AM237 activated TRPC5:C5 channels, with greater effect at positive voltages, but with lower efficacy than EA. Pico145 competitively inhibited AM237-induced TRPC5:C5 activation. AM237 did not activate TRPC4:C4, TRPC4-C1, TRPC5-C1, TRPC1:C5, and TRPC1:C4 channels, or native TRPC1:C4 channels in A498 cells, but potently inhibited EA-dependent activation of these channels with IC50 values ranging from 0.9 to 7 nM. AM237 (300 nM) did not activate or inhibit TRPC3, TRPC6, TRPV4, or TRPM2 channels. Conclusions and implications This study suggests the possibility for selective activation of TRPC5 channels by xanthine derivatives and supports the general principle that xanthine-based compounds can activate, potentiate, or inhibit these channels depending on subunit composition.
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- 2019
122. Over-Expression of TRPC6 via CRISPR Based Synergistic Activation Mediator in BMSCs Ameliorates Brain Injury in a Rat Model of Cerebral Ischemia/Reperfusion
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Jing Wang, Fan Yang, Wen-Bin Li, Yu-Shi Tang, Yu-Jun Pan, and Jin-Xing Gao
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Male ,0301 basic medicine ,Stromal cell ,Cell Survival ,Ischemia ,Pharmacology ,Brain Ischemia ,TRPC6 ,03 medical and health sciences ,Transient receptor potential channel ,0302 clinical medicine ,Mediator ,Animals ,Medicine ,Clustered Regularly Interspaced Short Palindromic Repeats ,Rats, Wistar ,Cyclic AMP Response Element-Binding Protein ,TRPC ,TRPC Cation Channels ,Neurons ,Brain-derived neurotrophic factor ,Calpain ,business.industry ,Brain-Derived Neurotrophic Factor ,General Neuroscience ,Spectrin ,Mesenchymal Stem Cells ,medicine.disease ,Neuroprotection ,Rats ,Up-Regulation ,030104 developmental biology ,Reperfusion Injury ,Models, Animal ,business ,Reperfusion injury ,030217 neurology & neurosurgery - Abstract
Stroke is a major life-threatening and disabling disease with a restricted therapeutic approach. Bone marrow stromal cells (BMSCs) possess proliferative ability and a multi-directional differentiation potential, and secrete a range of trophic/growth factors that can protect neurons after cerebral ischemia/reperfusion. Transient receptor potential canonical (TRPC) is a family of non-selective channels permeable to Ca2+, with several functions including neuronal survival. Over-expression of TRPC6, a subtype of the TRPC family, was shown to protect neurons against cerebral ischemia/reperfusion injury. However, it remains unclear whether over-expression of TRPC6 in BMSCs can further reduce brain injury after ischemia/reperfusion. In the present study, we report that over-expression of TRPC6 via a CRISPR-based synergistic activation mediator in BMSCs provided a greater reduction of brain injury in a rat model of ischemia/reperfusion. Further, the improved neurofunctional outcomes were associated with increased TRPC6 and brain derived neurotrophic factor expression levels. Overall, these data suggest that TRPC6 over-expressing BMSCs may be a promising therapeutic agent for ischemic stroke.
- Published
- 2019
123. Acid sphingomyelinase – a regulator of canonical transient receptor potential channel 6 (TRPC6) activity
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Martin Reichel, Bernd Nürnberg, Fabian Schumacher, Aksana Andreyeva, Kristina Friedland, Anna Fejtova, Lian Ye, Cosima Rhein, Johannes Kornhuber, Burkhard Kleuser, Stefanie Zeitler, Gabriel Nowak, and Juliana Monti
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0301 basic medicine ,Ceramide ,Medizin ,Ceramides ,PC12 Cells ,Biochemistry ,FIASMA ,TRPC6 ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Transient receptor potential channel ,chemistry.chemical_compound ,0302 clinical medicine ,ddc:570 ,medicine ,Animals ,Institut für Biochemie und Biologie ,Ion channel ,TRPC Cation Channels ,Neurons ,Rats ,Cell biology ,Sphingomyelin Phosphodiesterase ,030104 developmental biology ,chemistry ,Lipid modification ,Acid sphingomyelinase ,Sphingomyelin ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Recent investigations propose the acid sphingomyelinase (ASM)/ceramide system as a novel target for antidepressant action. ASM catalyzes the breakdown of the abundant membrane lipid sphingomyelin to the lipid messenger ceramide. This ASM‐induced lipid modification induces a local shift in membrane properties, which influences receptor clustering and downstream signaling. Canonical transient receptor potential channels 6 (TRPC6) are non‐selective cation channels located in the cell membrane that play an important role in dendritic growth, synaptic plasticity and cognition in the brain. They can be activated by hyperforin, an ingredient of the herbal remedy St. John’s wort for treatment of depression disorders. Because of their role in the context of major depression, we investigated the crosstalk between the ASM/ceramide system and TRPC6 ion channels in a pheochromocytoma cell line 12 neuronal cell model (PC12 rat pheochromocytoma cell line). Ca2+ imaging experiments indicated that hyperforin‐induced Ca2+ influx through TRPC6 channels is modulated by ASM activity. While antidepressants, known as functional inhibitors of ASM activity, reduced TRPC6‐mediated Ca2+ influx, extracellular application of bacterial sphingomyelinase rebalanced TRPC6 activity in a concentration‐related way. This effect was confirmed in whole‐cell patch clamp electrophysiology recordings. Lipidomic analyses revealed a decrease in very long chain ceramide/sphingomyelin molar ratio after ASM inhibition, which was connected with changes in the abundance of TRPC6 channels in flotillin‐1–positive lipid rafts as visualized by western blotting. Our data provide evidence that the ASM/ceramide system regulates TRPC6 channels likely by controlling their recruitment to specific lipid subdomains and thereby fine‐tuning their physical properties.
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- 2019
124. Ibudilast attenuates doxorubicin‐induced cytotoxicity by suppressing formation of TRPC3 channel and NADPH oxidase 2 protein complexes
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Koji Uchida, Kazuhiro Nishiyama, Yasu-Taka Azuma, Akiyuki Nishimura, Tomohiro Tanaka, Chiemi Toyama, Tomohiro Yamashita, Satoru Koyanagi, Yuko Ibuki, Takuro Numaga-Tomita, Naoya Matsunaga, Yasuyuki Fujimoto, Shigehiro Ohdo, and Motohiro Nishida
- Subjects
Male ,0301 basic medicine ,Cell Survival ,Pyridines ,Antineoplastic Agents ,Ibudilast ,Pharmacology ,Cell Line ,Rats, Sprague-Dawley ,Mice ,03 medical and health sciences ,0302 clinical medicine ,TRPC3 ,Western blot ,medicine ,Animals ,Humans ,Myocytes, Cardiac ,Doxorubicin ,Cytotoxicity ,TRPC Cation Channels ,Cardiotoxicity ,NADPH oxidase ,medicine.diagnostic_test ,biology ,Wasting Syndrome ,Chemistry ,Research Papers ,Mice, Inbred C57BL ,030104 developmental biology ,NADPH Oxidase 2 ,Toxicity ,biology.protein ,Tobacco Smoke Pollution ,Reactive Oxygen Species ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Background and purpose Doxorubicin is a highly effective anticancer agent but eventually induces cardiotoxicity associated with increased production of ROS. We previously reported that a pathological protein interaction between TRPC3 channels and NADPH oxidase 2 (Nox2) contributed to doxorubicin-induced cardiac atrophy in mice. Here we have investigated the effects of ibudilast, a drug already approved for clinical use and known to block doxorubicin-induced cytotoxicity, on the TRPC3-Nox2 complex. We specifically sought evidence that this drug attenuated doxorubicin-induced systemic tissue wasting in mice. Experimental approach We used the RAW264.7 macrophage cell line to screen 1,271 clinically approved chemical compounds, evaluating functional interactions between TRPC3 channels and Nox2, by measuring Nox2 protein stability and ROS production, with and without exposure to doxorubicin. In male C57BL/6 mice, samples of cardiac and gastrocnemius muscle were taken and analysed with morphometric, immunohistochemical, RT-PCR and western blot methods. In the passive smoking model, cells were exposed to DMEM containing cigarette sidestream smoke. Key results Ibudilast, an anti-asthmatic drug, attenuated ROS-mediated muscle toxicity induced by doxorubicin treatment or passive smoking, by inhibiting the functional interactions between TRPC3 channels and Nox2, without reducing TRPC3 channel activity. Conclusions and implications These results indicate a common mechanism underlying induction of systemic tissue wasting by doxorubicin. They also suggest that ibudilast could be repurposed to prevent muscle toxicity caused by anticancer drugs or passive smoking.
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- 2019
125. Peripheral myelin protein 22 modulates store-operated calcium channel activity, providing insights into Charcot-Marie-Tooth disease etiology
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Masayoshi Sakakura, Carlos G. Vanoye, Jun Li, Malathi Narayan, Charles R. Sanders, Bruce D. Carter, Rose M. Follis, and Alexandra J. Trevisan
- Subjects
0301 basic medicine ,Schwann cell ,Gadolinium ,Endoplasmic Reticulum ,Store-operated calcium channel activity ,Biochemistry ,TRPC1 ,Mice ,03 medical and health sciences ,Calcium imaging ,Neurobiology ,Charcot-Marie-Tooth Disease ,Peripheral myelin protein 22 ,medicine ,Animals ,Humans ,Stromal Interaction Molecule 1 ,Evoked Potentials ,Molecular Biology ,TRPC Cation Channels ,Mice, Knockout ,SOC channels ,030102 biochemistry & molecular biology ,Chemistry ,Endoplasmic reticulum ,STIM1 ,Cell Biology ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Mutagenesis, Site-Directed ,Calcium ,Calcium Channels ,Schwann Cells ,Myelin Proteins - Abstract
Charcot-Marie-Tooth (CMT) disease is a peripheral neuropathy associated with gene duplication and point mutations in the peripheral myelin protein 22 (PMP22) gene. However, the role of PMP22 in Schwann cell physiology and the mechanisms by which PMP22 mutations cause CMT are not well-understood. On the basis of homology between PMP22 and proteins associated with modulation of ion channels, we hypothesized that PMP22 alters ion channel activity. Using whole-cell electrophysiology, we show here that heterologous PMP22 expression increases the amplitude of currents similar to those ascribed to store-operated calcium (SOC) channels, particularly those involving transient receptor canonical channel 1 (TrpC1). These channels help replenish Ca(2+) in the endoplasmic reticulum (ER) following stimulus-induced depletion. Currents with similar properties were recorded in WT but not pmp22(−/−) mouse Schwann cells. Heterologous expression of the CMT-associated PMP22_L16P variant, which fails to reach the plasma membrane and localizes to the ER, led to larger currents than WT PMP22. Similarly, Schwann cells isolated from Trembler J (TrJ; PMP22_L16P) mice had larger currents than WT littermates. Calcium imaging in live nerves and cultured Schwann cells revealed elevated intracellular Ca(2+) in TrJ mice compared with WT. Moreover, we found that PMP22 co-immunoprecipitated with stromal interaction molecule 1 (STIM1), the Ca(2+) sensor SOC channel subunit in the ER. These results suggest that in the ER, PMP22 interacts with STIM1 and increases Ca(2+) influx through SOC channels. Excess or mutant PMP22 in the ER may elevate intracellular Ca(2+) levels, which could contribute to CMT pathology.
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- 2019
126. Trpc5 deficiency causes hypoprolactinemia and altered function of oscillatory dopamine neurons in the arcuate nucleus
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Ana Moreno-Pérez, Trese Leinders-Zufall, Petra Weissgerber, Thomas Blum, Frank Zufall, Bernd Bufe, Marc Freichel, Martina Pyrski, and Anela Arifovic
- Subjects
endocrine system ,medicine.medical_specialty ,Lactation Disorders ,Biology ,TRPC5 ,Membrane Potentials ,Mice ,Anterior pituitary ,Dopamine ,Internal medicine ,medicine ,Animals ,Homeostasis ,Humans ,hypothalamus ,TRPC Cation Channels ,Feedback, Physiological ,Multidisciplinary ,Dopaminergic Neurons ,Reproduction ,Arcuate Nucleus of Hypothalamus ,Genetic Diseases, Inborn ,Prolactin deficiency ,medicine.disease ,Trpc5 channelopathy ,Prolactin ,Hypoprolactinemia ,Endocrinology ,medicine.anatomical_structure ,Gene Expression Regulation ,PNAS Plus ,Hypothalamus ,Mutation ,HC-070 ,Female ,dopamine ,Arousal ,Gonadotropins ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction ,medicine.drug - Abstract
Dopamine neurons of the hypothalamic arcuate nucleus (ARC) tonically inhibit the release of the protein hormone prolactin from lactotropic cells in the anterior pituitary gland and thus play a central role in prolactin homeostasis of the body. Prolactin, in turn, orchestrates numerous important biological functions such as maternal behavior, reproduction, and sexual arousal. Here, we identify the canonical transient receptor potential channel Trpc5 as an essential requirement for normal function of dopamine ARC neurons and prolactin homeostasis. By analyzing female mice carrying targeted mutations in the Trpc5 gene including a conditional Trpc5 deletion, we show that Trpc5 is required for maintaining highly stereotyped infraslow membrane potential oscillations of dopamine ARC neurons. Trpc5 is also required for eliciting prolactin-evoked tonic plateau potentials in these neurons that are part of a regulatory feedback circuit. Trpc5 mutant females show severe prolactin deficiency or hypoprolactinemia that is associated with irregular reproductive cyclicity, gonadotropin imbalance, and impaired reproductive capabilities. These results reveal a previously unknown role for the cation channel Trpc5 in prolactin homeostasis of female mice and provide strategies to explore the genetic basis of reproductive disorders and other malfunctions associated with defective prolactin regulation in humans.
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- 2019
127. Focused ultrasound activates voltage-gated calcium channels through depolarizing TRPC1 sodium currents in kidney and skeletal muscle
- Author
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Tsang-Wei Tu, Joseph A. Frank, Scott R. Burks, Matthew E. Nagle, and Rebecca M. Lorsung
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0301 basic medicine ,Transient Receptor Potential Channel 1 ,Mechanotransduction ,Medicine (miscellaneous) ,Kidney ,TRPC1 ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Animals ,Muscle, Skeletal ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Ion channel ,Tropism ,Calcium signaling ,TRPC Cation Channels ,Ultrasonography ,Mice, Knockout ,Mice, Inbred C3H ,Voltage-Gated Calcium Channels ,Cavitation ,Voltage-dependent calcium channel ,Biological signal transduction ,Chemistry ,Sodium ,Mesenchymal Stem Cells ,030104 developmental biology ,Ultrasonic Waves ,Cyclooxygenase 2 ,Acoustic radiation force ,Biophysics ,focused ultrasound ,Mechanosensitive channels ,Calcium ,Female ,Depolarization ,Calcium Channels ,030217 neurology & neurosurgery ,Research Paper - Abstract
Pulsed focused ultrasound (pFUS) technology is being developed for clinical neuro/immune modulation and regenerative medicine. Biological signal transduction of pFUS forces can require mechanosensitive or voltage-gated plasma membrane ion channels. Previous studies suggested pFUS is capable of activating either channel type, but their mechanistic relationship remains ambiguous. We demonstrated pFUS bioeffects increased mesenchymal stem cell tropism (MSC) by altering molecular microenvironments through cyclooxygenase-2 (COX2)-dependent pathways. This study explored specific relationships between mechanosensitive and voltage-gated Ca2+ channels (VGCC) to initiate pFUS bioeffects that increase stem cell tropism. Methods: Murine kidneys and hamstring were given pFUS (1.15 or 1.125 MHz; 4MPa peak rarefactional pressure) under ultrasound or magnetic resonance imaging guidance. Cavitation and tissue displacement were measure by hydrophone and ultrasound radiofrequency data, respectively. Elastic modeling was performed from displacement measurements. COX2 expression and MSC tropism were evaluated in the presence of pharmacological ion channel inhibitors or in transient-receptor-potential-channel-1 (TRPC1)-deficient mice. Immunohistochemistry and co-immunoprecipitation examined physical channel relationships. Fluorescent ionophore imaging of cultured C2C12 muscle cells or TCMK1 kidney cells probed physiological interactions. Results: pFUS induced tissue deformations resulting in kPa-scale forces suggesting mechanical activation of pFUS-induced bioeffects. Inhibiting VGCC or TRPC1 in vivo blocked pFUS-induced COX2 upregulation and MSC tropism to kidneys and muscle. A TRPC1/VGCC complex was observed in plasma membranes. VGCC or TRPC1 suppression blocked pFUS-induced Ca2+ transients in TCMK1 and C2C12 cells. Additionally, Ca2+ transients were blocked by reducing transmembrane Na+ potentials and observed Na+ transients were diminished by genetic TRPC1 suppression. Conclusion: This study suggests that pFUS acoustic radiation forces mechanically activate a Na+-containing TRPC1 current upstream of VGCC rather than directly opening VGCC. The electrogenic function of TRPC1 provides potential mechanistic insight into other pFUS techniques for physiological modulation and optimization strategies for clinical implementation.
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- 2019
128. TRPC1 and ORAI1 channels in colon cancer
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Miriam Hernández-Morales, Lucía Núñez, Lucía Gutiérrez, Carlos Villalobos, Asociación Española Contra el Cáncer, Ministerio de Economía y Competitividad (España), Junta de Castilla y León, and Universidad de Valladolid
- Subjects
0301 basic medicine ,Stromal cell ,ORAI1 Protein ,Carcinogenesis ,Physiology ,Colorectal cancer ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Polyamines ,medicine ,Animals ,Humans ,Calcium Signaling ,Molecular Biology ,TRPC Cation Channels ,Chemistry ,Cell growth ,ORAI1 ,Cancer ,Cell Biology ,medicine.disease ,digestive system diseases ,Mitochondria ,030104 developmental biology ,Colonic Neoplasms ,Cancer cell ,Cancer research ,Calcium ,Store-operated Ca(2+) entry ,030217 neurology & neurosurgery ,Homeostasis - Abstract
Colon cancer cells, like other types of cancer cells, undergo the remodeling of the intracellular Ca2+ homeostasis that contributes to cancer cell hallmarks including enhanced cell proliferation, migration, and survival. Colon cancer cells display enhanced store-operated Ca2+ entry (SOCE) compared with their non-cancer counterparts. Colon cancer cells display an abnormal expression of SOCE molecular players including Orai1 and TRPC1 channels, and the stromal interacting molecule (STIM) 1 and 2. Interestingly, upregulation of Orai1 and TRPC1 channels and their contribution to SOCE are associated with cancer malignancy in colon cancer cells. In a specific cellular model of colon cancer, whereas in non-cancer colon cells SOCE is composed of the Ca2+ release activated (CRAC) currents, in colon cancer cells SOCE is composed of CRAC- and cationic, non-selective store operated (SOC) currents. Former SOCs are mediated by TRPC1 channels. Moreover, colon cancer cells also display dysregulation of the expression of 1,4,5-triphosphate receptors (IP3R) that could contribute to the enhanced SOCE. Another important factor underlying the enhanced SOCE is the differential mitochondrial modulation of the CRAC and SOC currents in non-cancer and colon cancer cells. In colon cancer cells, mitochondria take up more Ca2+ that prevent the Ca2+-dependent inactivation of the SOCs, leading to sustained Ca2+ entry. Notably, the inhibition of SOCE in cancer colon cells abolishes their cancer hallmarks. Robust evidence has shown the efficiency of non-steroidal anti-inflammatory drugs (NSAIDs) and difluoromethylornithine (DFMO) to reverse the enhanced cell proliferation, migration, and apoptosis resistance of cancer cells. In colon cancer cells, both NSAIDs and DFMO decrease SOCE, but they target different molecular components of SOCE. NSAIDs decrease the Ca2+ uptake by mitochondria, limiting their ability to prevent the Ca2+-dependent inactivation of the SOCs that underlie SOCE. On the other hand, DFMO inhibits the expression of TRPC1 channels in colon cancer cells, eliminating their contribution to SOCE. The identification of players of SOCE in colon cancer cells may help to better understand the remodeling of the Ca2+ homeostasis in cancer. Importantly, the use of different pharmacological tools that target different SOCE molecular players in colon cancer cells may play a pivotal role in designing better chemoprevention strategies., We gratefully acknowledge support from the Spanish Ministry of Economy and Competitivity (grant BFU2015-70131R), Junta de Castilla y León, Spain (grant VA294P18), the Spanish Association Against Cancer (AECC), and a predoctoral fellowship of the University of Valladolid to LGG.
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- 2019
129. TRPC3-Nox2 axis mediates nutritional deficiency-induced cardiomyocyte atrophy
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Tomohiro Tanaka, Kazuhiro Nishiyama, Takuro Numaga-Tomita, Supachoke Mangmool, Motohiro Nishida, Sayaka Oda, Akiyuki Nishimura, Suhaini Binti Sudi, and Caroline Sunggip
- Subjects
0301 basic medicine ,Programmed cell death ,Cell Survival ,lcsh:Medicine ,Models, Biological ,Article ,Contractility ,03 medical and health sciences ,Adenosine Triphosphate ,0302 clinical medicine ,Atrophy ,TRPC3 ,medicine ,Extracellular ,Animals ,Myocytes, Cardiac ,lcsh:Science ,TRPC Cation Channels ,Heart Failure ,chemistry.chemical_classification ,Reactive oxygen species ,Multidisciplinary ,NADPH oxidase ,biology ,Malnutrition ,lcsh:R ,NADPH Oxidases ,medicine.disease ,Adenosine ,Rats ,Cell biology ,030104 developmental biology ,chemistry ,NADPH Oxidase 2 ,biology.protein ,lcsh:Q ,Reactive Oxygen Species ,Biomarkers ,030217 neurology & neurosurgery ,Signal Transduction ,medicine.drug - Abstract
Myocardial atrophy, characterized by the decreases in size and contractility of cardiomyocytes, is caused by severe malnutrition and/or mechanical unloading. Extracellular adenosine 5′-triphosphate (ATP), known as a danger signal, is recognized to negatively regulate cell volume. However, it is obscure whether extracellular ATP contributes to cardiomyocyte atrophy. Here, we report that ATP induces atrophy of neonatal rat cardiomyocytes (NRCMs) without cell death through P2Y2 receptors. ATP led to overproduction of reactive oxygen species (ROS) through increased amount of NADPH oxidase (Nox) 2 proteins, due to increased physical interaction between Nox2 and canonical transient receptor potential 3 (TRPC3). This ATP-mediated formation of TRPC3-Nox2 complex was also pathophysiologically involved in nutritional deficiency-induced NRCM atrophy. Strikingly, knockdown of either TRPC3 or Nox2 suppressed nutritional deficiency-induced ATP release, as well as ROS production and NRCM atrophy. Taken together, we propose that TRPC3-Nox2 axis, activated by extracellular ATP, is the key component that mediates nutritional deficiency-induced cardiomyocyte atrophy.
- Published
- 2019
130. Triple-negative breast cancer cell line sensitivity to englerin A identifies a new, targetable subtype
- Author
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Susan L. Mooberry, Shayne D. Hastings, Madesh Muniswamy, Corena V Grant, Chase M. Carver, April L. Risinger, John A. Beutler, and Karthik Ramachandran
- Subjects
0301 basic medicine ,Cancer Research ,Sulforhodamine B ,Gene Expression ,Triple Negative Breast Neoplasms ,Article ,TRPC1 ,Mice ,Sesquiterpenes, Guaiane ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Cation homeostasis ,Animals ,Humans ,RNA, Small Interfering ,TRPC ,Triple-negative breast cancer ,TRPC Cation Channels ,Dose-Response Relationship, Drug ,Chemistry ,Depolarization ,Antineoplastic Agents, Phytogenic ,Mitochondria ,030104 developmental biology ,Oncology ,Drug Resistance, Neoplasm ,Cell culture ,Gene Knockdown Techniques ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Intracellular - Abstract
PURPOSE: Triple-negative breast cancers (TNBCs) represent a heterogeneous group of tumors. The lack of targeted therapies combined with the inherently aggressive nature of TNBCs results in a higher relapse rate and poorer overall survival. We evaluated the heterogeneity of TNBC cell lines for TRPC channel expression and sensitivity to cation-disrupting drugs. METHODS: The TRPC1/4/5 agonist englerin A was used to identify a group of TNBC cell lines sensitive to TRPC1/4/5 activation and intracellular cation disruption. Quantitative RT-PCR, the sulforhodamine B assay, pharmacological inhibition, and siRNA-mediated knockdown approaches were employed. Epifluorescence imaging was performed to measure intracellular Ca(2+) and Na(+) levels. Mitochondrial membrane potential changes were monitored by confocal imaging. RESULTS: BT-549 and Hs578T cells express high levels of TRPC4 and TRPC1/4, respectively, and are exquisitely, 2000- and 430-fold, more sensitive to englerin A than other TNBC cell lines. While englerin A caused a slow Na(+) and nominal Ca(2+) accumulation in Hs578T cells, it elicited rapid increases in cytosolic Ca(2+) levels that triggered mitochondrial depolarization in BT-549 cells. Interestingly, BT-549 and Hs578T cells were also more sensitive to digoxin as compared to other TNBC cell lines. Collectively, these data reveal TRPC1/4 channels as potential biomarkers of TNBC cell lines with dysfunctional mechanisms of cation homeostasis and therefore sensitivity to cardiac glycosides. CONCLUSIONS: The sensitivity of BT-549 and Hs578T cells to englerin A and digoxin suggests a subset of TNBCs are highly susceptible to cation disruption and encourages investigation of TRPC1 and TRPC4 as potential new biomarkers of sensitivity to cardiac glycosides.
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- 2019
131. TRPC1 as a negative regulator for TRPC4 and TRPC5 channels
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Chansik Hong, Jongyun Myeong, Juyeon Ko, Insuk So, Jinsung Kim, and Misun Kwak
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Neurons ,0301 basic medicine ,Voltage-dependent calcium channel ,Physiology ,Chemistry ,Clinical Biochemistry ,Brain ,TRPC5 ,TRPC4 ,Membrane Potentials ,TRPC1 ,03 medical and health sciences ,Transient receptor potential channel ,030104 developmental biology ,0302 clinical medicine ,TRPC3 ,Physiology (medical) ,Biophysics ,Animals ,Humans ,Homomeric ,Protein Multimerization ,030217 neurology & neurosurgery ,TRPC ,TRPC Cation Channels - Abstract
Transient receptor potential canonical (TRPC) channels are calcium permeable, non-selective cation channels with wide tissue-specific distribution. Among 7 TRPC channels, TRPC 1/4/5 and TRPC3/6/7 are subdivided based on amino acid sequence homology. TRPC4 and TRPC5 channels exhibit cationic current with homotetrameric form, but they also form heterotetrameric channel such as TRPC1/4 or TRPC1/5 once TRPC1 is incorporated. The expression of TRPC1 is ubiquitous whereas the expressions of TRPC4 and TRPC5 are rather focused in nervous system. With the help of conditional knock-out of TPRC1, 4 and/or 5 genes, TRPC channels made of these constituents are reported to be involved in various pathophysiological functions such as seizure, anxiety-like behaviour, fear, Huntington's disease, Parkinson's disease and many others. In heterologous expression system, many issues such as activation mechanism, stoichiometry and relative cation permeabilites of homomeric or heteromeric channels have been addressed. In this review, we discussed the role of TRPC1 channel per se in plasma membrane, role of TRPC1 in heterotetrameric conformation (TRPC1/4 or TRPC1/5) and relationship between TRPC1/4/5 channels, calcium influx and voltage-gated calcium channels.
- Published
- 2019
132. Expression and localization of transient receptor potential channels in the bovine uterus epithelium throughout the estrous cycle
- Author
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Sadegh Shirian, Maryam Ghavideldarestani, Alexandra E. Butler, and Stephen L. Atkin
- Subjects
0301 basic medicine ,Gene isoform ,Immunocytochemistry ,Estrous Cycle ,Biology ,Endometrium ,Epithelium ,TRPC6 ,TRPC1 ,03 medical and health sciences ,Transient receptor potential channel ,Transient Receptor Potential Channels ,0302 clinical medicine ,Genetics ,medicine ,Animals ,Protein Isoforms ,Molecular Biology ,TRPC ,TRPC Cation Channels ,Uterus ,General Medicine ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cattle ,Female ,Transcriptome - Abstract
Transient receptor potential (TRP) channels are expressed in the endometrium but it is unknown if they are modulated through the estrous cycle (EC). This study was undertaken to identify the modulation of the TRPC gene and protein isoforms in bovine uterine epithelium, as a model for human, throughout the EC. Changes in the expression of TRPC genes in bovine uterine epithelium throughout the EC were measured using Real-Time PCR, while immunohistochemistry and immunocytochemistry were used to determine the localization of these channels. Out of the 7 members of the TRPC family, TRPC1, 2, 3, 4 and 6 genes were expressed in bovine uterine epithelial tissue and TRPC 5 and 7 were not. Gene expression levels of all TRPC isoforms underwent cyclical changes throughout the EC. Moreover, cyclical changes were detected in the protein levels of TRPC1 and TRPC6 throughout the EC. These findings show that TRPC channels are modulated through the EC and therefore may have a role in reproductive events.
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- 2019
133. Transcription Factor 2I Regulates Neuronal Development via TRPC3 in 7q11.23 Disorder Models
- Author
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Zhong Ping Feng, Alexander J. Groffen, Ya Chi Huang, Hong Shuo Sun, Michael Wu, Jacqueline N. Crawley, Lucy R. Osborne, Ekaterina Turlova, Nardos G. Tassew, Wenliang Chen, Marielle Deurloo, You Wei Lin, Philippe P. Monnier, Elaine Tam, Human genetics, Amsterdam Neuroscience - Cellular & Molecular Mechanisms, and Functional Genomics
- Subjects
0301 basic medicine ,Cortical neurons ,Time Factors ,Neurons/metabolism ,TRPC3 ,Calcium in biology ,Transcription Factors, TFII ,Mice ,0302 clinical medicine ,Gene duplication ,Williams-Beuren syndrome (WBS) ,Neurons ,General transcription factor ,Phenotype ,TRPC Cation Channels/metabolism ,Cell biology ,Transcription Factors, TFII/metabolism ,Neurology ,TFII/metabolism ,Williams syndrome ,Neurites/metabolism ,congenital, hereditary, and neonatal diseases and abnormalities ,Calcium/metabolism ,Neuroscience (miscellaneous) ,Context (language use) ,Axons/metabolism ,Biology ,Article ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,SDG 3 - Good Health and Well-being ,medicine ,Neurites ,Animals ,Transcription factor ,TRPC Cation Channels ,Chromosome Aberrations ,General transcription factor 2i ,Animal ,Cell Membrane/metabolism ,Cell Membrane ,medicine.disease ,Axons ,Disease Models, Animal ,030104 developmental biology ,Disease Models ,Calcium ,030217 neurology & neurosurgery ,Transcription Factors - Abstract
Williams syndrome (WS) and 7q11.23 duplication syndrome (Dup7q11.23) are neurodevelopmental disorders caused by the deletion and duplication, respectively, of ~ 25 protein-coding genes on chromosome 7q11.23. The general transcription factor 2I (GTF2I, protein TFII-I) is one of these proteins and has been implicated in the neurodevelopmental phenotypes of WS and Dup7q11.23. Here, we investigated the effect of copy number alterations in Gtf2i on neuronal maturation and intracellular calcium entry mechanisms known to be associated with this process. Mice with a single copy of Gtf2i (Gtf2i+/Del) had increased axonal outgrowth and increased TRPC3-mediated calcium entry upon carbachol stimulation. In contrast, mice with 3 copies of Gtf2i (Gtf2i+/Dup) had decreases in axon outgrowth and in TRPC3-mediated calcium entry. The underlying mechanism was that TFII-I did not affect TRPC3 protein expression, while it regulated TRPC3 membrane translocation. Together, our results provide novel functional insight into the cellular mechanisms that underlie neuronal maturation in the context of the 7q11.23 disorders. Electronic supplementary material The online version of this article (10.1007/s12035-018-1290-7) contains supplementary material, which is available to authorized users.
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- 2019
134. Treasure troves of pharmacological tools to study transient receptor potential canonical 1/4/5 channels
- Author
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Hussein N. Rubaiy
- Subjects
Pharmacology ,Drug discovery ,Chemistry ,TRPC5 ,TRPC4 ,Riluzole ,TRPC1 ,Transient receptor potential channel ,medicine ,Animals ,Humans ,Review Articles ,Neuroscience ,Ion channel ,TRPC ,TRPC Cation Channels ,medicine.drug - Abstract
Canonical or classical transient receptor potential 4 and 5 proteins (TRPC4 and TRPC5) assemble as homomers or heteromerize with TRPC1 protein to form functional nonselective cationic channels with high calcium permeability. These channel complexes, TRPC1/4/5, are widely expressed in nervous and cardiovascular systems, also in other human tissues and cell types. It is debatable that TRPC1 protein is able to form a functional ion channel on its own. A recent explosion of molecular information about TRPC1/4/5 has emerged including knowledge of their distribution, function, and regulation suggesting these three members of the TRPC subfamily of TRP channels play crucial roles in human physiology and pathology. Therefore, these ion channels represent potential drug targets for cancer, epilepsy, anxiety, pain, and cardiac remodelling. In recent years, a number of highly selective small-molecule modulators of TRPC1/4/5 channels have been identified as being potent with improved pharmacological properties. This review will focus on recent remarkable small-molecule agonists: (-)-englerin A and tonantzitlolone and antagonists: Pico145 and HC7090, of TPRC1/4/5 channels. In addition, this work highlights other recently identified modulators of these channels such as the benzothiadiazine derivative, riluzole, ML204, clemizole, and AC1903. Together, these treasure troves of agonists and antagonists of TRPC1/4/5 channels provide valuable hints to comprehend the functional importance of these ion channels in native cells and in vivo animal models. Importantly, human diseases and disorders mediated by these proteins can be studied using these compounds to perhaps initiate drug discovery efforts to develop novel therapeutic agents.
- Published
- 2019
135. TRPC channels mediated calcium entry is required for proliferation of human airway smooth muscle cells induced by nicotine-nAChR
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Chao Li, Ling Chen, Aiguo Dai, Gongyong Peng, Xing Yang, Pixin Ran, Wen Li, Yongliang Jiang, Lei Liu, Heshen Tian, Dan Pan, and Yumin Zhou
- Subjects
Male ,0301 basic medicine ,Nicotine ,Small interfering RNA ,Myocytes, Smooth Muscle ,Respiratory System ,chemistry.chemical_element ,Receptors, Nicotinic ,Calcium ,Biochemistry ,Pulmonary Disease, Chronic Obstructive ,03 medical and health sciences ,TRPC3 ,Calcium imaging ,medicine ,Humans ,Calcium Signaling ,Viability assay ,TRPC ,Aged ,Cell Proliferation ,TRPC Cation Channels ,030102 biochemistry & molecular biology ,Chemistry ,General Medicine ,Middle Aged ,Cell biology ,Nicotinic acetylcholine receptor ,030104 developmental biology ,Gene Expression Regulation ,Female ,medicine.drug - Abstract
The present study was designed to explore the role of transient receptor potential canonical 3 (TRPC3) in nicotine-induced chronic obstructive pulmonary disease (COPD) and its underlying mechanism. In this study, the expression and localization of α5 nicotinic acetylcholine receptor (α5-nAchR) in lung tissues were determined by western blotting and immunohistochemistry. The quantitative real-time PCR (qRT-PCR) analysis was performed to examine the mRNA expression levels of α5-nAchR and TRPC3 in human airway smooth muscle cells (HASMCs). Cell viability was assessed by CCK-8 assay. Proliferation was detected by cell counting and EdU immunofluorescent staining. Fluorescence calcium imaging was carried out to measure cytosolic Ca2+ ([Ca2+]cyt) concentration. The results showed that the α5-nAchR and TRPC3 expressions were significantly up-regulated in lung tissues of COPD smokers. Nicotine promoted HASMC proliferation, which was accompanied by elevated α5-nAchR and TRPC3 expressions, basal [Ca2+]cyt, store-operated calcium entry (SOCE) and the rate of Mn2+ quenching in HASMCs. Further investigation indicated that nicotine-induced Ca2+ response and TRPC3 up-regulation was reversibly blocked by small interfering RNA (siRNA) suppression of α5-nAChR. The knockdown of TRPC3 blunted Ca2+ response and HASMC proliferation induced by nicotine. In conclusion, nicotine-induced HASMC proliferation was mediated by TRPC3-dependent calcium entry via α5-nAchR, which provided a potential target for treatment of COPD.
- Published
- 2019
136. Switching between persistent firing and depolarization block in individual rat CA1 pyramidal neurons
- Author
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Beate Knauer and Motoharu Yoshida
- Subjects
Male ,Potassium Channels ,Patch-Clamp Techniques ,antagonists & inhibitors [TRPC Cation Channels] ,physiology [Electrophysiological Phenomena] ,Cognitive Neuroscience ,pharmacology [Muscarinic Agonists] ,metabolism [TRPC Cation Channels] ,drug effects [Pyramidal Cells] ,Hippocampus ,Stimulation ,Muscarinic Agonists ,In Vitro Techniques ,Hippocampal formation ,050105 experimental psychology ,Membrane Potentials ,pharmacology [Carbachol] ,03 medical and health sciences ,0302 clinical medicine ,Current clamp ,Animals ,Rats, Long-Evans ,0501 psychology and cognitive sciences ,ddc:610 ,Patch clamp ,CA1 Region, Hippocampal ,TRPC Cation Channels ,physiology [CA1 Region, Hippocampal] ,Dose-Response Relationship, Drug ,physiology [Pyramidal Cells] ,Chemistry ,Pyramidal Cells ,05 social sciences ,cytology [CA1 Region, Hippocampal] ,drug effects [Membrane Potentials] ,Depolarization ,metabolism [Potassium Channels] ,drug effects [Electrophysiological Phenomena] ,Potassium channel ,Electrophysiological Phenomena ,Rats ,drug effects [CA1 Region, Hippocampal] ,Cholinergic ,Carbachol ,Female ,Neuroscience ,030217 neurology & neurosurgery - Abstract
The hippocampal formation plays a role in mnemonic tasks and epileptic discharges in vivo. In vitro, these functions and malfunctions may relate to persistent firing (PF) and depolarization block (DB), respectively. Pyramidal neurons of the CA1 field have previously been reported to engage in either PF or DB during cholinergic stimulation. However, it is unknown whether these cells constitute disparate populations of neurons. Furthermore, it is unclear which cell-specific peculiarities may mediate their diverse response properties. However, it has not been shown whether individual CA1 pyramidal neurons can switch between PF and DB states. Here, we used whole cell patch clamp in the current clamp mode on in vitro CA1 pyramidal neurons from acutely sliced rat tissue to test various intrinsic properties which may provoke individual cells to switch between PF and DB. We found that individual cells could switch from PF to DB, in a cholinergic agonist concentration dependent manner and depending on the parameters of stimulation. We also demonstrate involvement of TRPC and potassium channels in this switching. Finally, we report that the probability for DB was more pronounced in the proximal than in the distal half of CA1. These findings offer a potential mechanism for the stronger spatial modulation in proximal, compared to distal CA1, as place field formation was shown to be affected by DB. Taken together, our results suggest that PF and DB are not mutually exclusive response properties of individual neurons. Rather, a cell's response mode depends on a variety of intrinsic properties, and modulation of these properties enables switching between PF and DB.
- Published
- 2019
137. Combination of leflunomide and benazepril reduces renal injury of diabetic nephropathy rats and inhibits high-glucose induced cell apoptosis through regulation of NF-κB, TGF-β and TRPC6
- Author
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Huanhuan Yang, Fang Tian, Juzhen Yan, Hui-li Li, Zhangqing Zhou, and Yuanyuan Wang
- Subjects
Blood Glucose ,Male ,benazepril ,030232 urology & nephrology ,TRPC6 ,Apoptosis ,030204 cardiovascular system & hematology ,Critical Care and Intensive Care Medicine ,NF-κB ,Rats, Sprague-Dawley ,Diabetic nephropathy ,chemistry.chemical_compound ,0302 clinical medicine ,Transforming Growth Factor beta ,Laboratory Study ,Diabetic Nephropathies ,Blood urea nitrogen ,Leflunomide ,Kidney ,NF-kappa B ,Drug Synergism ,General Medicine ,medicine.anatomical_structure ,Nephrology ,Mesangial Cells ,Drug Therapy, Combination ,Glomerular Filtration Rate ,Signal Transduction ,medicine.drug ,TGF-β ,medicine.medical_specialty ,Renal function ,Benazepril ,Streptozocin ,Cell Line ,Diabetes Mellitus, Experimental ,03 medical and health sciences ,Internal medicine ,medicine ,Animals ,Humans ,TRPC Cation Channels ,Creatinine ,business.industry ,diabetic nephropathy ,Benzazepines ,Streptozotocin ,medicine.disease ,Diseases of the genitourinary system. Urology ,Rats ,Endocrinology ,chemistry ,Hyperglycemia ,RC870-923 ,business - Abstract
Objective: To investigate effects of combination use of leflunomide and benazepril on diabetic nephropathy (DN) both in vivo and in vitro. Methods: The streptozotocin (STZ) induced Sprague-Dawley rats were treated with leflunomide (15 mg/kg/d), benazepril (15 mg/kg/d) or both the two drugs. Fasting blood glucose (FBG) and renal function indexes including blood urea nitrogen (BUN), serum creatinine (Scr), and proteinuria and kidney/body weight ratio (KW/BW) were measured. HE staining was used for histological analysis. The rat glomerular mesangial cells (RMCs) were treated with high-glucose (150 mg/ml) and the leflunomide and benazepril with both concentrations of 50 μmol/l were used to treat the high-glucose induced cells. TUNEL assay was used for measurement of cell apoptosis. Western blotting was conducted to determine expression of nuclear factor Kappa B (NF-κB), transforming growth factor-β (TGF-β) and transient receptor potential canonical 6 (TRPC6). Results: The body weight was significantly lower and all indexes of FBG, BUN, Scr, proteinuria and KW/BW ratio, GFR, as well as inflammatory factors TNF-α and IL-6 were significantly increased in the DN group after STZ treatment for 4 weeks. The treatment with leflunomide, benazepril or the both dramatically reduced the above effects induced by STZ, and the alteration was the most significant in the combination group. Treatment of leflunomide and benazepril significantly reduced expression levels of NF-κB, TGF-β and TRPC6 in renal tissues of DN rats as well as in high-glucose induced RMCs. It was also observed leflunomide and benazepril reduced high-glucose induced cell apoptosis of RMCs. Conclusion: The combination use of leflunomide and benazepril could improve the renal function and reduce the renal injury of DN rats and could reduce the levels of NF-κb, TGF-β and TRPC6 in both DN rats and high-glucose induced RMCs.
- Published
- 2019
138. Design, synthesis and characterization of novel N-heterocyclic-1-benzyl-1H-benzo[d]imidazole-2-amines as selective TRPC5 inhibitors leading to the identification of the selective compound, AC1903
- Author
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Swagat Sharma, Mónica S. Montesinos, Anna Greka, Corey R. Hopkins, and Juan Pablo
- Subjects
Benzimidazole ,Indazoles ,Clinical Biochemistry ,Pharmaceutical Science ,TRPC5 ,01 natural sciences ,Biochemistry ,Article ,Podocyte ,Structure-Activity Relationship ,chemistry.chemical_compound ,Transient receptor potential channel ,Heterocyclic Compounds ,Drug Discovery ,medicine ,Animals ,Humans ,Imidazole ,Amines ,Renal Insufficiency, Chronic ,Molecular Biology ,TRPC Cation Channels ,Kidney ,Dose-Response Relationship, Drug ,Molecular Structure ,010405 organic chemistry ,Organic Chemistry ,Imidazoles ,medicine.disease ,Combinatorial chemistry ,0104 chemical sciences ,Disease Models, Animal ,010404 medicinal & biomolecular chemistry ,medicine.anatomical_structure ,chemistry ,Design synthesis ,Drug Design ,Molecular Medicine ,Kidney disease - Abstract
The transient receptor potential cation channel 5 (TRPC5) has been previously shown to affect podocyte survival in the kidney. As such, inhibitors of TRPC5 are interesting candidates for the treatment of chronic kidney disease (CKD). Herein, we report the synthesis and biological characterization of a series of N-heterocyclic-1-benzyl-1H-benzo[d]imidazole-2-amines as selective TRPC5 inhibitors. Work reported here evaluates the benzimidazole scaffold and substituents resulting in the discovery of AC1903, a TRPC5 inhibitor that is active in multiple animal models of CKD.
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- 2019
139. The TRPC6 inhibitor, larixyl acetate, is effective in protecting against traumatic brain injury-induced systemic endothelial dysfunction
- Author
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B. Paul Herring, Ashley M. Riley, Alexander G. Obukhov, Fletcher A. White, Xingjuan Chen, and Natalie N. Taylor-Nguyen
- Subjects
Lipopolysaccharides ,0301 basic medicine ,Aorta, Thoracic ,Vascular permeability ,Acetates ,lcsh:RC346-429 ,Cerebral circulation ,Mice ,0302 clinical medicine ,Traumatic brain injury ,Head Injuries, Closed ,Brain Injuries, Traumatic ,Endothelial dysfunction ,TRPC ,Aorta ,Mice, Knockout ,General Neuroscience ,3. Good health ,Vasodilation ,TRPC channels ,Neuroprotective Agents ,Neurology ,Cerebral blood flow ,Knockout mouse ,medicine.medical_specialty ,Immunology ,Naphthalenes ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Isometric Contraction ,Internal medicine ,TRPC6 Cation Channel ,medicine ,Animals ,Vascular Diseases ,Neuroinflammation ,lcsh:Neurology. Diseases of the nervous system ,TRPC Cation Channels ,business.industry ,Research ,medicine.disease ,Mice, Inbred C57BL ,Toll-Like Receptor 4 ,030104 developmental biology ,Endocrinology ,Endothelium, Vascular ,business ,030217 neurology & neurosurgery - Abstract
Background The incidence of traumatic brain injuries (TBIs) is on the rise in the USA. Concussions, or mild TBIs without skull fracture, account for about 75% of all TBIs. Mild TBIs (mTBIs) lead to memory and cognitive deficits, headaches, intraocular pressure rises, axonal degeneration, neuroinflammation, and an array of cerebrovascular dysfunctions, including increased vascular permeability and decreased cerebral blood flow. It has been recently reported that besides vascular dysfunction in the cerebral circulation, mTBI may also cause a significant impairment of endothelial function in the systemic circulation, at least within mesenteric microvessels. In this study, we investigated whether mTBI affects endothelial function in aortas and determined the contribution of transient receptor potential canonical (TRPC) channels to modulating mTBI-associated endothelial dysfunction. Methods We used a model of closed-head mTBI in C57BL/6, 129S, 129S-C57BL/6-F2 mice, and 129S-TRPC1 and 129S-C57BL/6-TRPC6 knockout mice to determine the effect of mTBI on endothelial function in mouse aortas employing ex vivo isometric tension measurements. Aortic tissue was also analyzed using immunofluorescence and qRT-PCR for TRPC6 expression following mTBI. Results We show that in various strains of mice, mTBI induces a pronounced and long-lasting endothelial dysfunction in the aorta. Ablation of TRPC6 protects mice from mTBI-associated aortic endothelial dysfunction, while TRPC1 ablation does not impact brain injury-induced endothelial impairment in the aorta. Consistent with a role of TRPC6 activation following mTBI, we observed improved endothelial function in wild type control mice subjected to mTBI following 7-day in vivo treatment with larixyl acetate, an inhibitor of TRPC6 channels. Conversely, in vitro treatment with the pro-inflammatory endotoxin lipopolysaccharide, which activates endothelial TRPC6 in a Toll-like receptor type 4 (TLR4)-dependent manner, worsened aortic endothelial dysfunction in wild type mice. Lipopolysaccharide treatment in vitro failed to elicit endothelial dysfunction in TRPC6 knockout mice. No change in endothelial TRPC6 expression was observed 7 days following TBI. Conclusions These data suggest that TRPC6 activation may be critical for inducing endothelial dysfunction following closed-head mTBI and that pharmacological inhibition of the channel may be a feasible therapeutic strategy for preventing mTBI-associated systemic endothelial dysfunction.
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- 2019
140. The TRPC5 channel regulates angiogenesis and promotes recovery from ischemic injury in mice
- Author
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Mengru Gao, Xiaoqiang Yao, Mingxu Xie, Tingting Zhou, Xin Ma, Yifei Zhu, Lei Feng, Wing Tak Wong, and Aiqin Mao
- Subjects
0301 basic medicine ,Angiogenesis ,Cell ,Ischemia ,TRPC5 ,Biochemistry ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Retinal Diseases ,medicine ,Animals ,Humans ,Therapeutic angiogenesis ,Molecular Biology ,TRPC Cation Channels ,Mice, Knockout ,Riluzole ,Neovascularization, Pathologic ,030102 biochemistry & molecular biology ,business.industry ,Endothelial Cells ,Cell Biology ,medicine.disease ,Vascular endothelial growth factor ,Endothelial stem cell ,Disease Models, Animal ,HEK293 Cells ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Cancer research ,business ,medicine.drug - Abstract
Ischemia-related diseases are a leading cause of death worldwide, and promoting therapeutic angiogenesis is key for effective recovery from hypoxia-ischemia. Given the limited success of angiogenic factors, such as vascular endothelial growth factor, in clinical trials, it is important to find more promising angiogenic targets. Here, using both cell- and tissue-based assays and a mouse model of injury-induced ischemia, we investigated the involvement of the transient receptor potential canonical 5 (TRPC5) ion channel in angiogenesis and the effects of a TRPC5 activator, the Food and Drug Administration-approved drug riluzole, on recovery from ischemic injury. We demonstrate that TRPC5 is involved in endothelial cell sprouting, angiogenesis, and blood perfusion in an oxygen-induced retinopathy model and a hind limb ischemia model. We found a potential regulatory link between nuclear factor of activated T cell isoform c3 and angiopoietin-1 that could provide the mechanistic basis for the angiogenic function of TRPC5. Importantly, treatment with riluzole, which can activate TRPC5 in endothelial cells, improved recovery from ischemia in mice. Our study reveals TRPC5 as a potential angiogenic target and suggests riluzole as a promising drug for managing ischemic diseases.
- Published
- 2019
141. Inhibition of TRPC1 prevents cardiac hypertrophy via NF-κB signaling pathway in human pluripotent stem cell-derived cardiomyocytes
- Author
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Jiaxi Shen, Tongyu Li, Tingyu Gong, Xiaochen Wang, Ling Tang, Bing Dai, Jue Wang, Li Wang, Danni Zhou, Fengfeng Guo, Jun Su, Ping Liang, Hao Wang, Fang Yao, Hongkun Wang, and Haodi Wu
- Subjects
Pluripotent Stem Cells ,0301 basic medicine ,Cardiomegaly ,030204 cardiovascular system & hematology ,Muscle hypertrophy ,TRPC1 ,03 medical and health sciences ,chemistry.chemical_compound ,Transient receptor potential channel ,0302 clinical medicine ,medicine ,Humans ,Myocytes, Cardiac ,Induced pluripotent stem cell ,Molecular Biology ,TRPC Cation Channels ,Base Sequence ,business.industry ,NF-kappa B ,Hypertrophic cardiomyopathy ,NF-κB ,medicine.disease ,Cell biology ,030104 developmental biology ,chemistry ,Heart failure ,Phorbol ,Cardiology and Cardiovascular Medicine ,business ,Signal Transduction - Abstract
Cardiac hypertrophy is an adaptive response against increased workload featuring by an increase in left ventricular mass and a thickening left ventricle wall. Here, we showed the expression of transient receptor potential canonical 1 (TRPC1) is higher in hearts of patients with hypertrophic cardiomyopathy (HCM) or heart failure (HF) than that of normal hearts. To better understand the mechanisms of TRPC1 in regulating cellular hypertrophy of human-based cardiomyocytes, we generated human pluripotent stem cell lines of TRPC1 knockout by CRISPR/Cas9. We demonstrated that knockout of TRPC1 significantly attenuated cardiomyocyte hypertrophy phenotype induced by phorbol 12-myristate 13-acetate, which was associated with abnormal activation of NF-κB. In contrast, overexpression of TRPC1 induced cardiomyocyte hypertrophy, which can be reversed by inhibition of NF-κB. Taken together, we established a stable human-based cardiomyocyte hypertrophy model and highlighted molecular mechanisms underlying TRPC1-mediated hypertrophy, aiding the development of therapeutic drugs for HCM and HF by targeting TRPC1.
- Published
- 2019
142. TRPC6 Inactivation Reduces Albuminuria Induced by Protein Overload in Sprague Dawley Rats
- Author
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Eun Young, Kim and Stuart E, Dryer
- Subjects
Rats, Sprague-Dawley ,Disease Models, Animal ,Glomerular Filtration Barrier ,Albumins ,Albuminuria ,Animals ,Nephritis, Interstitial ,General Medicine ,TRPC6 ,TRPC5 ,glomerular physiology ,tubulointerstitial fibrosis ,albuminuria ,TRPC Cation Channels - Abstract
Canonical transient receptor potential-6 (TRPC6) channels have been implicated in familial and acquired forms of focal and segmental glomerulosclerosis (FSGS), and in renal fibrosis following ureteral obstruction in mice. TRPC6 channels also appear to play a role in driving glomerular disease in aging and in autoimmune glomerulonephritis. In the present study, we examine the role of TRPC6 in the proteinuric state caused by prolonged albumin overload (AO) in Sprague Dawley rats induced by daily injections of exogenous albumin. This was assessed in rats with a global and constitutive inactivation of TRPC6 channels (Trpc6del/del rats) and in wild-type littermates (Trpc6wt/wt rats). AO for 14 and 28 days caused increased urine albumin excretion that was significantly attenuated in Trpc6del/del rats compared to Trpc6wt/wt controls. AO overload did not induce significant glomerulosclerosis or azotemia in either genotype. AO induced mild tubulointerstitial disease characterized by fibrosis, hypercellularity and increased expression of markers of fibrosis and inflammation. Those changes were equally severe in Trpc6wt/wt and Trpc6del/del rats. Immunoblot analysis of renal cortex indicated that AO increased the abundances of TRPC3 and TRPC6, and caused a nearly complete loss of TRPC5 in Trpc6wt/wt rats. The increase in TRPC3 and the loss of TRPC5 occurred to the same extent in Trpc6del/del rats. These data also suggest that TRPC6 plays a role in the normal function of the glomerular filtration barrier. However, whether TRPC6 inactivation protects the tubulointerstitial compartments in Sprague Dawley rats depends on the disease model examined.
- Published
- 2022
143. Protein detection and localization of the non-selective cation channel TRPC6 in the human heart
- Author
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Tobias, Jacobs, Jan, Abdinghoff, and Thomas, Tschernig
- Subjects
Pharmacology ,Cardiovascular Diseases ,Cations ,TRPC6 Cation Channel ,Animals ,Humans ,Heart ,Ion Channels ,TRPC Cation Channels - Abstract
Due to longer lifespans in societies in industrialized countries, cardiovascular diseases are becoming increasingly important for medical research. It has already been shown that the cell membrane-bound, non-selective TRPC6 ion channel is important in the pathogenesis of heart diseases. Among other things, it is permeable to calcium ion, which plays a critical role in cardiac contraction and relaxation. The TRPC6 ion channel is a promising therapeutic target in the treatment of cardiovascular diseases. A deeper understanding of the physiological and pathophysiological role as well as the localization of TRPC6 in human cardiac tissue is the basis for new drug development. Although the TRPC6 channel has been detected in animal studies, at the mRNA level in humans, and sparse TRPC6 protein has been detected in humans, there are no systematic studies of TRPC6 protein detection in the human heart. For the first time, TRPC6 ion channel protein was detected histologically in human heart tissue from body donors in different structures, localizations, and histological layers - particularly in cardiomyocytes and intramuscular arterioles - by immunohistochemistry, just as TRPC6 expression has already been shown in animal models of the heart by other research groups. In the sense of the translational concept, this indicates a possible transferability of research results from animal models to humans.
- Published
- 2022
144. TRPV4 and TRPC1 channels mediate the response to tensile strain in mouse Müller cells
- Author
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Andrew O. Jo, Monika Lakk, Christopher N. Rudzitis, and David Križaj
- Subjects
Mammals ,Physiology ,Ependymoglial Cells ,TRPV Cation Channels ,Cell Biology ,Ion Channels ,Article ,Mice ,Animals ,Calcium ,Calcium Channels ,Calcium Signaling ,Molecular Biology ,TRPC Cation Channels - Abstract
Müller glia, a pillar of metabolic, volume regulatory and immune/inflammatory signaling in the mammalian retina, are among the earliest responders to mechanical stressors in the eye. Ocular trauma, edema, detachment and glaucoma evokes early inflammatory activation of Müller cells yet the identity of their mechanotransducers and their downstream signaling mechanisms remain unknown. Here, we investigate expression of genes that encode putative stretch-activated calcium channels (SACs) in mouse Müller cells together with their response to dynamical tensile loading in cells loaded with a calcium indicator dye. Transcript levels in purified glia were Trpc1>Piezo1>Trpv2>Trpv4>>Trpv1>Trpa1. Cyclic radial deformation of matrix -coated substrates produced dose-dependent increases in [Ca(2+)](i) that were suppressed by the TRPV4 channel antagonist HC-067047 and by ablation of the Trpv4 gene. In addition, stretch-evoked calcium responses were reduced by knockdown and pharmacological inhibition of TRPC1 channels whereas the TRPV2 inhibitor tranilast had no effect. These data demonstrate that Müller cells are intrinsically mechanosensitive, with the response to tensile loading mediated through synergistic activation of TRPV4 and TRPC1 channels. Coupling between mechanical stress and Müller Ca(2+) homeostasis has treatment implications, since many neuronal injury paradigms in the retina involve calcium dysregulation associated with inflammatory and immune signaling.
- Published
- 2022
145. Magnolol and honokiol target TRPC4 to regulate extracellular calcium influx and relax intestinal smooth muscle
- Author
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Lin Niu, Jie Wang, Fukui Shen, Jie Gao, Min Jiang, and Gang Bai
- Subjects
Male ,Pharmacology ,Biphenyl Compounds ,Myocytes, Smooth Muscle ,TRPV Cation Channels ,Muscle, Smooth ,Lignans ,Rats ,Intestines ,Rats, Sprague-Dawley ,HEK293 Cells ,Drug Discovery ,Animals ,Humans ,Calcium Signaling ,Medicine, Chinese Traditional ,Muscle Contraction ,TRPC Cation Channels - Abstract
Magnolia officinalis Cortex (M. officinalis) is a classical traditional Chinese medicine (TCM) widely used to treat digestive system diseases. It effectively regulates gastrointestinal motility to improve abdominal pain, abdominal distension and other symptoms. Magnolol (MAG) and honokiol (HON) are the main pharmacodynamic components responsible for the gastrointestinal activity of M. officinalis.The transient receptor potential (TRP) family is highly expressed in the gastrointestinal tract and participates in the regulation of gastrointestinal motility, visceral hypersensitivity, visceral secretion and other physiological activities. In this study, the calcium-lowering mechanisms of MAG and HON contributing to the smooth muscle relaxation associated with TRP are discussed.The relaxation smooth muscle effects of MAG and HON were tested by the isolated intestine tone tests. A synthetic MAG probe (MAG-P) was used to target fishing for their possible target. The distribution of MAG on the smooth muscle was identified by a molecular tracer based on chemical biology. CaAfter confirming the smooth muscle relaxation in the small intestine induced by MAG and HON, the relaxation effect was identified mainly due to the downregulation of intracellular calcium by controlling external calcium influx. Although MAG and HON inhibited both TRPV4 and TRPC4 channels to reduce calcium levels, the inhibitory effect on TRPC4 channels is an important mechanism of their smooth muscle relaxation effect, since TRPC4 is widely expressed in the small intestinal smooth muscle cells.The inhibition of MAG and HON on TRPC4 channels contributes to the relaxation of intestinal smooth muscle.
- Published
- 2022
146. Discovery of pyrroledione analogs as potent transient receptor potential canonical channel 5 inhibitors
- Author
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Zhuang, Zhang, Lili, Chen, Hongtao, Tian, Mengru, Liu, Shan, Jiang, Jianhua, Shen, Kai, Wang, and Zhengyu, Cao
- Subjects
Dose-Response Relationship, Drug ,Molecular Structure ,Podocytes ,Organic Chemistry ,Clinical Biochemistry ,Pharmaceutical Science ,Biochemistry ,Structure-Activity Relationship ,Drug Discovery ,Humans ,Molecular Medicine ,Pyrroles ,Protamines ,Molecular Biology ,TRPC Cation Channels - Abstract
A deepening understanding of the relationship between transient receptor potential canonical channel 5 (TRPC5) and chronic kidney disease (CKD), has led to the emergence of several types of TRPC5 inhibitors displaying clear therapeutic effect. Herein, we report the synthesis and biological evaluation of a series of pyrroledione TRPC5 inhibitors, culminating in the discovery of compound 16g with subtype selectivity. Compared with GFB-8438, a potent TRPC5 inhibitor (Goldfinch Bio), compound 16g showed improved inhibition of TRPC5 and enhanced protective effect against protamine sulfates (PS)-induced podocyte injury in vitro. In addition, compound 16g did not induce cell death in primary cultured hepatocytes and immortalized podocytes in a preliminary toxicity assessment, indicating its utility as a potent and safe inhibitor for studying the function of TRPC5.
- Published
- 2022
147. TRPC1 Deficiency Exacerbates Cerebral Ischemia/Reperfusion-Induced Neurological Injury by Potentiating Nox4-Derived Reactive Oxygen Species Generation
- Author
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Yingli Feng, Hao Meng, Honglei Wang, Ning Xu, Tianyi Liu, and Yuan Qi
- Subjects
0301 basic medicine ,Mitochondrial ROS ,medicine.medical_specialty ,TRPC1 ,Physiology ,Down-Regulation ,Ischemia/reperfusion injury ,Protein degradation ,medicine.disease_cause ,lcsh:Physiology ,Brain Ischemia ,Cell Line ,Superoxide dismutase ,Neurological dysfunction ,lcsh:Biochemistry ,03 medical and health sciences ,Mice ,Nox4 ,0302 clinical medicine ,Internal medicine ,medicine ,NADPH ,Animals ,lcsh:QD415-436 ,TRPC Cation Channels ,chemistry.chemical_classification ,Mice, Knockout ,Reactive oxygen species ,NADPH oxidase ,biology ,lcsh:QP1-981 ,NOX4 ,Oxidative Stress ,030104 developmental biology ,Endocrinology ,chemistry ,NADPH Oxidase 4 ,Reperfusion Injury ,biology.protein ,Calcium ,P22phox ,030217 neurology & neurosurgery ,Oxidative stress - Abstract
Background/Aims: Transient receptor potential cation channel 1 (TRPC1)-mediated the calcium (Ca2+) influx plays an important role in several brain disorders. However, the function of TRPC1 in ischemia/reperfusion (I/R)-induced neurological injury is unclear. Methods: Wild-type or TRPC1 knockout mice underwent middle cerebral artery occlusion for 90 min followed by 24 h of reperfusion. In an in vitro study, neuronal cells were treated with oxygen–glucose deprivation and reoxygenation (OGD/R) to mimic I/R. The intracellular Ca2+ concentration [Ca2+]i was measured by Fura 2-AM under a microscope. Cerebral infarct volume was measured by triphenyltetrazolium chloride staining. Neurological function was examined by neurological severity score, Morris water maze test, rotarod test and string test. Oxidative parameters were detected by malondialdehyde, glutathione peroxidase, and superoxide dismutase commercially available kits. The protein expression levels of TRPC1, Nox4, p22phox, p47phox, and p67phox were analyzed by western blotting. Results: Brain tissues from cerebral I/R mice showed decreased TRPC1 expression. Similarly, TRPC1 expression was reduced in HT22 cells upon exposure to OGD/R treatment, followed by decreased Ca2+ influx. However, TRPC1 overexpression reversed the OGD/R-induced decrease in [Ca2+]i. TRPC1 knockout significantly exacerbated I/R-induced brain infarction, edema, neurological severity score, memory impairment, neurological deficits, and oxidative stress. In contrast, TRPC1 upregulation inhibited the increase in reactive oxygen species (ROS) generation induced by OGD/R. Analysis of key subunits of the Nox family and mitochondrial ROS revealed that the effects of TRPC1 downregulation on oxidative stress were associated with activation of Nox4-containing NADPH oxidase. TRPC1 interacted with Nox4 and facilitated Nox4 protein degradation under OGD/R conditions. In addition, TRPC1 inhibition potentiated the OGD/R-induced translocation of p47phox and p67phox as well as the interaction between Nox4 and p47phox or p67phox, whereas TRPC1 overexpression had the opposite effects. Conclusion: TRPC1 deficiency potentiates ROS generation via Nox4-containing NADPH oxidase, which exacerbates cerebral I/R injury. TRPC1 may be a promising molecular target for the treatment of stroke.
- Published
- 2018
148. Topotecan prevents hypoxia-induced pulmonary arterial hypertension and inhibits hypoxia-inducible factor-1α and TRPC channels
- Author
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Yongliang Jiang, Ling Chen, Gongyong Peng, Xing Yang, Yumin Zhou, Nian Liu, Pixin Ran, Wen Li, Lei Liu, Chao Li, Aiguo Dai, Heshen Tian, and Dan Pan
- Subjects
Male ,Vascular Endothelial Growth Factor A ,0301 basic medicine ,Hypertension, Pulmonary ,Myocytes, Smooth Muscle ,Vascular Remodeling ,Pharmacology ,Biochemistry ,Muscle hypertrophy ,TRPC1 ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Cell Movement ,In vivo ,medicine ,Animals ,Rats, Wistar ,TRPC ,Cell Proliferation ,TRPC Cation Channels ,Chemistry ,Cell Biology ,Hypoxia (medical) ,Hypoxia-Inducible Factor 1, alpha Subunit ,Cell Hypoxia ,Rats ,Phenotype ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Hypoxia-inducible factors ,Ventricle ,030220 oncology & carcinogenesis ,Cytokines ,Calcium ,medicine.symptom ,Topotecan - Abstract
Background This study aimed to investigate the effects of topotecan (TPT) on the hypoxia-induced pulmonary arterial hypertension (PAH) in a rat model, and to explore the underlying mechanism. Methods The experiments were carried out in vitro using rat PASMCs and in vivo using a rat model of hypoxia-induced PAH. Results TPT significantly suppressed the hypoxia-induced upregulation of HIF-1α and TRPC1/4/6 expression both in pulmonary arterial smooth muscle cells (PASMCs) from normal rats and in pulmonary arteries from PAH model rats. Furthermore, TPT effectively inhibited intracellular Ca2+ concentration ([Ca2+]i) change (Ca2+ influx) in PASMCs from both normal rats and PAH model rats. Importantly, TPT treatment significantly inhibited the hypoxia-induced proliferation, migration and a contractile-to-synthetic phenotypic switching of normal rat PASMCs in vitro, where the effect was abrogated by overexpression of TRPC1/4/6. Furthermore, TPT administration potently attenuated the hypoxia-induced PAH-associated pulmonary arteriolar remodeling in PAH model rats, as evidenced by amelioration of elevated hemodynamic parameters, and enhanced right ventricle hypertrophy and wall thickening. Conclusion TPT ameliorates the hypoxia-induced pulmonary vascular remodeling in PAH, and the mechanism is associated with TPT-mediated inhibition of hypoxia-induced upregulation of HIF-1α and TRPC1/4/6 expression, Ca2+ influx, and PASMCs proliferation.
- Published
- 2018
149. TRPC1 regulates brown adipose tissue activity in a PPARγ-dependent manner
- Author
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Christian Wolfrum, Pawel Pelczar, and Elke Kiehlmann
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Physiology ,Endocrinology, Diabetes and Metabolism ,Down-Regulation ,TRPV Cation Channels ,Peroxisome proliferator-activated receptor ,Weight Gain ,TRPC1 ,Mice ,03 medical and health sciences ,Transient receptor potential channel ,chemistry.chemical_compound ,Adipose Tissue, Brown ,Downregulation and upregulation ,Physiology (medical) ,Internal medicine ,Adipocyte ,Brown adipose tissue ,medicine ,Animals ,Uncoupling Protein 1 ,TRPC Cation Channels ,Mice, Knockout ,chemistry.chemical_classification ,Gene knockdown ,Chemistry ,Thermogenin ,Up-Regulation ,PPAR gamma ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Calcium Channels ,Energy Metabolism - Abstract
Brown adipose tissue (BAT) has the unique ability to convert energy stored in the form of triglycerides into heat. This property makes BAT a target tissue to increase energy expenditure and improve systemic metabolic control. TRPC1 is a founding member of the TRP protein family that also includes several temperature sensitive channels. We show that TRPC1 is highly expressed in all adipocyte depots including BAT and that Trpc1-deficient mice are prone to weight gain and manifest reduced metabolic control. We also demonstrate that knockdown of TRPC1 in cultured brown adipocytes leads to a downregulation of several metabolic genes, including UCP1 and PPARγ, as well as upregulation of a BAT-specific thermosensitive channel TRPV2, ultimately resulting in impaired respiratory function. We also show that TRPC1 is a possible target of PPARγ, suggesting that TRPC1 is a downstream component of a mechanism that translates metabolic or environmental stimuli into output in the form of BAT activity. Better understanding of the possible role of TRPC1 and other TRP channels in body temperature regulation and BAT function may help us to develop obesity therapies based on BAT activation.
- Published
- 2018
150. TRPCing around the hypothalamus
- Author
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Jian Qiu, Oline K. Rønnekleiv, and Martin J. Kelly
- Subjects
0301 basic medicine ,Pro-Opiomelanocortin ,Hypothalamus ,Article ,Energy homeostasis ,Receptors, G-Protein-Coupled ,Gonadotropin-Releasing Hormone ,03 medical and health sciences ,Transient receptor potential channel ,Kisspeptin ,Animals ,Humans ,Insulin ,TRPC ,TRPC Cation Channels ,G protein-coupled receptor ,Neurons ,Orexins ,Endocrine and Autonomic Systems ,Chemistry ,Cell biology ,Orexin ,030104 developmental biology ,Metabotropic glutamate receptor ,Intercellular Signaling Peptides and Proteins - Abstract
All of the canonical transient receptor potential channels (TRPC) with the exception of TRPC 2 are expressed in hypothalamic neurons and are involved in multiple homeostatic functions. Although the metabotropic glutamate receptors have been shown to be coupled to TRPC channel activation in cortical and sub-cortical brain regions, in the hypothalamus multiple amine and peptidergic G protein-coupled receptors (GPCRs) and growth factor/cytokine receptors are linked to activation of TRPC channels that are vital for reproduction, temperature regulation, arousal and energy homeostasis. In addition to the neurotransmitters, circulating hormones like insulin and leptin through their cognate receptors activate TRPC channels in POMC neurons. Many of the post-synaptic effects of the neurotransmitters and hormones are regulated in different physiological states by expression of TRPC channels in the post-synaptic neurons. Therefore, TRPC channels are key targets not only for neurotransmitters but circulating hormones in their vital role to control multiple hypothalamic functions, which is the focus of this review.
- Published
- 2018
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