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Voltage- and Ca2+-dependent, slow-activating vacuolar (SV) channels represent the major cation conductance of the plant vacuole membrane and have been implicated in calcium-induced calcium release (CICR) from intracellular stores. Encoded by the TWO PORE CHANNEL 1 gene, the Arabidopsis thaliana SV channel AtTPC1 has become a paradigm in understanding the synergistic action of membrane potential and cytosolic Ca2+ transients for channel gating. Novel AtTPC1 structural data give rise to emerging models describing the intermediate conformational transitions during channel opening and the resulting functional consequences. Across phyla, voltage-gated ion channels (VGICs) allow excitability. The vacuolar two-pore channel AtTPC1 from the tiny mustard plant Arabidopsis thaliana has emerged as a paradigm for deciphering the role of voltage and calcium signals in membrane excitation. Among the numerous experimentally determined structures of VGICs, AtTPC1 was the first to be revealed in a closed and resting state, fueling speculation about structural rearrangements during channel activation. Two independent reports on the structure of a partially opened AtTPC1 channel protein have led to working models that offer promising insights into the molecular switches associated with the gating process. We review new structure–function models and also discuss the evolutionary impact of two-pore channels (TPCs) on K+ homeostasis and vacuolar excitability. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H2S) is considered to mediate plant growth and development. However, whether H2S regulates the adaptation of mangrove plant to intertidal flooding habitats is not well understood. In this study, sodium hydrosulfide (NaHS) was used as an H2S donor to investigate the effect of H2S on the responses of mangrove plant Avicennia marina to waterlogging. The results showed that 24‐h waterlogging increased reactive oxygen species (ROS) and cell death in roots. Excessive mitochondrial ROS accumulation is highly oxidative and leads to mitochondrial structural and functional damage. However, the application of NaHS counteracted the oxidative damage caused by waterlogging. The mitochondrial ROS production was reduced by H2S through increasing the expressions of the alternative oxidase genes and increasing the proportion of alternative respiratory pathway in the total mitochondrial respiration. Secondly, H2S enhanced the capacity of the antioxidant system. Meanwhile, H2S induced Ca2+ influx and activated the expression of intracellular Ca2+‐sensing‐related genes. In addition, the alleviating effect of H2S on waterlogging can be reversed by Ca2+ chelator and Ca2+ channel blockers. In conclusion, this study provides the first evidence to explain the role of H2S in waterlogging adaptation in mangrove plants from the mitochondrial aspect. Summary statement: H2S reduces mitochondrial ROS production by increasing the expressions of the alternative oxidase gene (AOX) and increases the proportion of alternative respiratory pathway in the total respiration, while H2S alleviates the mitochondrial structural damages including loss of bilayer membranes and blurring of mitochondrial cristae, and functional damages such as decrease of mitochondrial membrane potential (ΔΨm), membrane permeability transition pores opening, Cyt c release and the blockage of electron transfer along the mitochondrial electron transport chain caused by waterlogging‐induced oxidative stress. Secondly, H2S enhances the capacity of the antioxidant system through upregulating the expression of SOD, CAT, and APX genes to scavenge excess ROS. Last, H2S induces Ca2+ influx in the roots under waterlogging. Our results provide the better understanding of the role of H2S in adaptation of mangrove plants to the coastal intertidal environment from the mitochondrial aspect. [ABSTRACT FROM AUTHOR]

Throughout the last decades, neuromorphic circuits have incited the interest of scientists, as they are potentially a powerful tool for the treatment of neurological diseases. To this end, it is essential to consider the biological principles of the CNS and develop the appropriate area- and power-efficient circuits. Motivated by studies that outline the indispensable role of astrocytes in the dynamic regulation of synaptic transmission and their active contribution to neural information processing in the CNS, in this work we propose a digital implementation of neuron–astrocyte bidirectional interactions. In order to describe the neuronal dynamics and the astrocytes' calcium dynamics, a modified version of the original Izhikevich neuron model was combined with a linear approximation of the Postnov functional neural–glial interaction model. For the implementation of the neural–glial computation core, only three pipeline stages and a 10.10 fixed point representation were utilized. Regarding the results obtained from the FPGA implementation and the comparisons to other works, the proposed neural–glial circuit reported significant savings in area requirements (from 22.53% up to 164.20%) along with considerable savings in total power consumption of 28.07% without sacrificing output computation accuracy. Finally, an RMSE analysis was conducted, confirming that this particular implementation produces more accurate results compared to previous studies. [ABSTRACT FROM AUTHOR]

Cancer is the second leading cause of death worldwide after heart disease. The current treatment options to fight cancer are limited, and there is a critical need for better treatment strategies. During the last several decades, several electric field (EF)-based approaches for anti-cancer therapies have been introduced, such as electroporation and tumor-treating fields; still, they are far from optimal due to their invasive nature, limited efficacy and significant side effects. In this study, we developed a non-contact EF stimulation system to investigate the in vitro effects of a novel EF modality on cancer biomarkers in normal (human astrocytes, human pancreatic ductal epithelial -HDPE-cells) and cancer cell lines (glioblastoma U87-GBM, human pancreatic cancer cfPac-1, and MiaPaCa-2). Our results demonstrate that this EF modality can successfully modulate an important cancer cell biomarker-cell surface phosphatidylserine (PS). Our results further suggest that moderate, but not low, amplitude EF induces p38 mitogen-activated protein kinase (MAPK), actin polymerization, and cell cycle arrest in cancer cell lines. Based on our results, we propose a mechanism for EF-mediated PS exposure in cancer cells, where the magnitude of induced EF on the cell surface can differentially regulate intracellular calcium (Ca2+) levels, thereby modulating surface PS exposure. [ABSTRACT FROM AUTHOR]

Background and Purpose: α4β2 nicotinic ACh receptors (nAChRs) comprise the most abundant class of nAChRs in the nervous system. They assemble in two stoichiometric forms, each exhibiting distinct functional and pharmacological signatures. However, whether one or both forms are modulated by calcium or magnesium has not been established. Experimental Approach To assess the functional consequences of calcium and magnesium, each stoichiometric form was expressed in clonal mammalian fibroblasts and single‐channel currents were recorded in the presence of a range of ACh concentrations. Key Results: In the absence of divalent cations, each stoichiometric form exhibits high unitary conductance and simple gating kinetics composed of solitary channel openings or short bursts of openings. However, in the presence of calcium and magnesium, the conductance and gating kinetics change in a stoichiometry‐dependent manner. Calcium and magnesium reduce the conductance of both stoichiometric forms, with each cation producing an equivalent reduction, but the reduction is greater for the (α4)2(β2)3 form. Moreover, divalent cations promote efficient channel opening of the (α4)3(β2)2 stoichiometry, while minimally affecting the (α4)2(β2)3 stoichiometry. For the (α4)3(β2)2 stoichiometry, at high but not low ACh concentrations, calcium in synergy with magnesium promote clustering of channel openings into episodes of many openings in quick succession. Conclusion and Implications: Modulation of the α4β2 nAChR by divalent cations depends on the ACh concentration, the type of cation and the subunit stoichiometry. The functional consequences of modulation are expected to depend on the regional distributions of the stoichiometric forms and synaptic versus extrasynaptic locations of the receptors. [ABSTRACT FROM AUTHOR]

Cancer is the second leading cause of death worldwide after heart disease. The current treatment options to fight cancer are limited, and there is a critical need for better treatment strategies. During the last several decades, several electric field (EF)-based approaches for anti-cancer therapies have been introduced, such as electroporation and tumor-treating fields; still, they are far from optimal due to their invasive nature, limited efficacy and significant side effects. In this study, we developed a non-contact EF stimulation system to investigate the in vitro effects of a novel EF modality on cancer biomarkers in normal (human astrocytes, human pancreatic ductal epithelial -HDPE-cells) and cancer cell lines (glioblastoma U87-GBM, human pancreatic cancer cfPac-1, and MiaPaCa-2). Our results demonstrate that this EF modality can successfully modulate an important cancer cell biomarker-cell surface phosphatidylserine (PS). Our results further suggest that moderate, but not low, amplitude EF induces p38 mitogen-activated protein kinase (MAPK), actin polymerization, and cell cycle arrest in cancer cell lines. Based on our results, we propose a mechanism for EF-mediated PS exposure in cancer cells, where the magnitude of induced EF on the cell surface can differentially regulate intracellular calcium (Ca2+) levels, thereby modulating surface PS exposure.

Neurophysiological observations are clarifying how astrocytes can actively participate in information processing and how they can encode information through frequency and amplitude modulation of intracellular Ca2+ signals. Consequently, hardware realization of astrocytes is important for developing the next generation of bio-inspired computing systems. In this paper, astrocytic calcium oscillations and neuronal firing dynamics are presented by De Pittà and IF (Integrated & Fire) models, respectively. Considering highly nonlinear equations of the astrocyte model, linear approximation and single constant multiplication (SCM) techniques are employed for efficient hardware execution while maintaining the dynamic of the original models. This low-cost hardware architecture for the astrocyte model is able to show the essential features of different types of Ca2+ modulation such as amplitude modulation (AM), frequency modulation (FM), or both modes (AFM). To show good agreement between the results of original models simulated in MATLAB and the proposed digital circuits executed on FPGA, quantitative, and qualitative analyses including phase plane are done. This new neuromorphic circuit of astrocyte is able to successfully demonstrate AM/FM/AFM calcium signaling in its real operation on FPGA and has applications in self-repairing systems. It also can be employed as a subsystem for linking biological cells to artificial neuronal networks using astrocytic calcium oscillations in future research.

Neurophysiological observations are clarifying how astrocytes can actively participate in information processing and how they can encode information through frequency and amplitude modulation of intracellular Ca2+ signals. Consequently, hardware realization of astrocytes is important for developing the next generation of bio-inspired computing systems. In this paper, astrocytic calcium oscillations and neuronal firing dynamics are presented by De Pittà and IF (Integrated & Fire) models, respectively. Considering highly nonlinear equations of the astrocyte model, linear approximation and single constant multiplication (SCM) techniques are employed for efficient hardware execution while maintaining the dynamic of the original models. This low-cost hardware architecture for the astrocyte model is able to show the essential features of different types of Ca2+ modulation such as amplitude modulation (AM), frequency modulation (FM), or both modes (AFM). To show good agreement between the results of original models simulated in MATLAB and the proposed digital circuits executed on FPGA, quantitative, and qualitative analyses including phase plane are done. This new neuromorphic circuit of astrocyte is able to successfully demonstrate AM/FM/AFM calcium signaling in its real operation on FPGA and has applications in self-repairing systems. It also can be employed as a subsystem for linking biological cells to artificial neuronal networks using astrocytic calcium oscillations in future research. [ABSTRACT FROM AUTHOR]

Abstract: In this work we describe a new family of dibenzo[1,4,5]thiadiazepines (2–12) that showed an interesting in vitro biological profile, namely neuroprotective and antioxidant properties, as well as blockade of cytosolic calcium entry. They showed no cytotoxic effects and the majority were predicted as CNS-permeable compounds. In human neuroblastoma cells they displayed good neuroprotective properties against mitochondrial oxidative stress which, in many cases, almost reached the full protection (>90%) when compounds were incubated with cells 24 h before the addition of toxic stressors. In co-incubation conditions these figures were smaller, although some compounds maintained an interesting level of neuroprotection, higher than 50%. Four selected compounds (2, 5, 8, and 11) were found to be effective antioxidant agents by sequestering mitochondrial radical oxygen species (ROS). Moreover, compound 2 showed a remarkable calcium-channel modulating activity. The interest of these compounds is increased by the fact that dibenzo[1,4,5]thiadiazepine is a barely known structure that is not difficult to synthesize and presents very few described derivatives, opening a new and broad line of research in Medicinal Chemistry. [Copyright &y& Elsevier]

Abstract: We have previously reported the multifunctional profile of N-(3-chloro-10H-phenothiazin-10-yl)-3-(dimethylamino)propanamide (1) as an effective neuroprotectant and selective butyrylcholinesterase inhibitor. In this paper, we have developed a series of N-acylaminophenothiazines obtained from our compound library or newly synthesised. At micro- and sub-micromolar concentrations, these compounds selectively inhibited butyrylcholinesterase (BuChE), protected neurons against damage caused by both exogenous and mitochondrial free radicals, showed low toxicity, and could penetrate into the CNS. In addition, N-(3-chloro-10H-phenothiazin-10-yl)-2-(pyrrolidin-1-yl)acetamide (11) modulated the cytosolic calcium concentration and protected human neuroblastoma cells against several toxics, such as calcium overload induced by an l-type Ca2+-channel agonist, tau-hyperphosphorylation induced by okadaic acid and Aβ peptide. [Copyright &y& Elsevier]

Abstract: The influence of 20nm ZnO nanoparticles on cytotoxicity, oxidative stress, intracellular calcium homeostasis, and gene expression was studied in human bronchial epithelial cells (BEAS-2B). ZnO caused a concentration- and time-dependent cytotoxicity while elevating oxidative stress and causing membrane damage (cellular LDH release). There was a remarkably steep relationship between concentration and toxicity at concentrations from 5 to 10μg/ml. Cytotoxicity was completely abolished by the antioxidant N-acetylcysteine (NAC). Exposure to ZnO also increased intracellular calcium levels ([Ca2+]in) in a concentration- and time-dependent manner that was partially attenuated by NAC. Nifedipine, a calcium channel blocker, partially attenuated the elevated [Ca2+]in, indicating that some of the excess [Ca2+]in is a result of influx from outside the cell. The relationships between oxidative stress, [Ca2+]in, and cytotoxicity are discussed. Exposure to a sublethal concentration of ZnO increased the expression of four genes that are involved in apoptosis and oxidative stress responses BNIP, PRDX3, PRNP, and TXRND1, by at least 2.5-fold. Thus, ZnO alters transcriptional regulation in BEAS-2B cells. [Copyright &y& Elsevier]

Background and Objectives: Curcuma caesia (family Zingiberaceae) is widely used in India as both an anti-inflammatory and anti-asthmatic in Ayurvedic medicine. However, there are no published pharmacological data on Curcuma caesia on its potential anti-asthmatic activity. Hence, the objective of the present investigation is to study the mechanisms by which the hydroalcoholic extract of Curcuma caesia relaxes the smooth muscle in the bronchioles and vasculature of the respiratory tract. Methods: The hydroalcoholic extract of Curcuma caesia (CC extract) was tested for its per se relaxant effect in guinea pig trachea and also in the presence of various receptor antagonists and enzyme inhibitors namely propranalol, 2′, 5′-dideoxyadenosine, methylene blue, glibenclamide, Nω-nitro-L-arginine(L-NNA) and α-chymotrypsin. Furthermore, the possible role of hydroalcoholic extract in calcium channel modulation was investigated in depolarized rabbit aorta. Results: The CC extract concentration dependently relaxed the carbachol (1 μM)-induced pre-contractions; the IC50 value was found to be 239.36 μg/ml and the incubation of either receptor antagonists or enzyme inhibitors did not exhibit any effect on the relaxation. In the isotonic Ca2+-free high-K+ (60 mM) depolarized aorta, CC extract (30 μg/mM) inhibited concentration-response curves of cumulative Ca2+ (0.1-30 mM) and the PD'2 value was found to be 4.11 μg/ml. Interpretation and Conclusion: The extract showed a dose-dependent, non-specific relaxation of pre-contracted isolated guinea pig trachea. The non-specific relaxant effect of the extract may be due to its ability to modulate calcium activity. [ABSTRACT FROM AUTHOR]

Abstract: A neuronal type Ca2+ stimulated nitric oxide synthase was earlier reported by us to be present in the protozoan parasite Leishmania donovani. As part of nitric oxide-cyclic GMP transduction signaling operative in higher eukaryotes and involved in the long-term potentiation, a soluble guanylyl cyclase has also been detected in this lower eukaryote. However, detailed biochemical characterization revealed the enzyme to be Ca2+ modulated and unstimulated by nitric oxide donors as opposed to higher eukaryotes. The possible role of intracellular Ca2+ level in the regulation of guanylyl cyclase activity as well as L. donovani infectivity was explored by measuring the intracellular survival of the parasites in mammalian macrophages after treatments, which decrease or elevate the intracellular Ca2+. Parasites loaded with intracellular Ca2+ chelators displayed significantly decreased infectivity and cyclic GMP level. In contrast, pretreatment with Ca2+ ionophores, which elevated Ca2+ levels in L. donovani, significantly enhanced the cyclic GMP level as well as the infectivity of the parasites. Moreover, treatment with selective inhibitors of soluble guanylyl cyclase also reduced infectivity, even in cases of calcium ionophore-treated parasites. The gene encoding the soluble guanylyl cyclase was cloned, sequenced and over expressed in bacterial system. The recombinant protein showed enzyme characteristics similar to that obtained in L. donovani promastigote cytosol. Together these results suggest a possible link between guanylyl cyclase, intracellular Ca2+ content and parasite infectivity. [Copyright &y& Elsevier]

An aminopeptidase secreted from Streptomyces septatus TH-2 (SSAP) was identified as a heat stable enzyme, and the Ssap gene was cloned and sequenced. The primary structure of SSAP showed 71% identity with that of a Streptomyces griseus aminopeptidase (SGAP), however, it lacked a unique calcium binding site. The recombinant SSAP was overexpressed in the culture supernatant of Escherichia coli harboring pET-KmS2. A comparison of recombinant SSAP and SGAP showed that both enzymes are different in terms of modulation by calcium and substrate specificity. The activity of SSAP was not modulated by calcium, while SGAP is a calcium-activated enzyme. SSAP catalyzed the hydrolysis of l-Lys-pNA efficiently whereas the reaction rate for l-Lys-pNA hydrolysis of SGAP was significantly low. Furthermore, in SGAP, the presence of Ca2+ decreased the reaction rate of l-Lys-pNA hydrolysis. SSAP also had different pKas of reaction from that of SGAP, although almost all the residues which compose the active site were conserved in both enzymes. This result indicates that SSAP has a different environment of substrate binding and active sites from those of SGAP. [Copyright &y& Elsevier]

The influence of GM1 on the neuritogenic phase of neuronal differentiation has been highlighted in recent reports showing upregulation of this ganglioside in the plasma and nuclear membranes concomitant with axonogenesis. These changes are accompanied by alterations in Ca2+ flux which constitute an essential component of the signaling mechanism for axon outgrowth. This study examines 2 distinct mechanisms of induced neurite outgrowth involving plasma membrane GM1, as expressed in 3 neuroblastoma cell lines. Growth of Neuro-2a and NG108-15 cells in the presence of neuraminidase (N'ase), an enzyme that increases the cell surface content of GM1, caused prolific outgrowth of neurites which, in the case of Neuro-2a, could be blocked by the B subunit of cholera toxin (Ctx B) which binds specifically to GM1; however, the latter agent applied to NG108-15 cells proved neuritogenic and potentiated the effect of N'ase. With N18 cells, the combination was also neuritogenic as was Ctx B alone, whereas N'ase by itself had no effect. Neurite outgrowth correlated with influx of extracellular Ca2+, determined with fura-2. Treatment of NG108-15 and N18 cells with Ctx B alone caused modest but persistent elevation of intracellular Ca2+ while a more pronounced increase occurred with the combination Ctx B + N'ase. Treatment with N'ase alone also caused modest but prolonged elevation of intracellular Ca2+ in NG108-15 and Neuro-2a but not N18; in the case of Neuro-2a this effect was blocked by Ctx B. Neuro-2a and N18 thus possess 2 distinctly different mechanisms for neuritogenesis based on Ca2+ modulation by plasma membrane GM1, while NG108-15 cells show both capabilities. The neurites stimulated by N'ase + Ctx B treatment of N18 cells were shown to have axonal character, as previously demonstrated for NG108-15 cells stimulated in this manner and for Neuro-2a cells stimulated by N'ase alone. [ABSTRACT FROM AUTHOR]

Abstract Background Plasmodium has a complex cell biology and it is essential to dissect the cell-signalling pathways underlying its survival within the host. Methods Using the fluorescence resonance energy transfer (FRET) peptide substrate Abz-AIKFFARQ-EDDnp and Fluo4/AM, the effects of extracellular ATP on triggering proteolysis and Ca2+ signalling in Plasmodium berghei and Plasmodium yoelii malaria parasites were investigated. Results The protease activity was blocked in the presence of the purinergic receptor blockers suramin (50 μM) and PPADS (50 μM) or the extracellular and intracellular calcium chelators EGTA (5 mM) and BAPTA/AM (25, 100, 200 and 500 μM), respectively for P. yoelii and P. berghei. Addition of ATP (50, 70, 200 and 250 μM) to isolated parasites previously loaded with Fluo4/AM in a Ca2+-containing medium led to an increase in cytosolic calcium. This rise was blocked by pre-incubating the parasites with either purinergic antagonists PPADS (50 μM), TNP-ATP (50 μM) or the purinergic blockers KN-62 (10 μM) and Ip5I (10 μM). Incubating P. berghei infected cells with KN-62 (200 μM) resulted in a changed profile of merozoite surface protein 1 (MSP1) processing as revealed by western blot assays. Moreover incubating P. berghei for 17 h with KN-62 (10 μM) led to an increase in rings forms (82% ± 4, n = 11) and a decrease in trophozoite forms (18% ± 4, n = 11). Conclusions The data clearly show that purinergic signalling modulates P. berghei protease(s) activity and that MSP1 is one target in this pathway.

Streptomyces griseus aminopeptidase is a zinc metalloenzyme containing 2 mol zinc/mol protein, similar to the homologous enzyme Aeromonas proteolytica aminopeptidase. In addition, a unique Ca2+-binding site has been identified in the Streptomyces enzyme, which is absent in the Aeromonas enzyme. Binding of Ca2+ enhances stability of the Streptomyces enzyme and modulates its activity and affinity towards substrates and inhibitors in a structure-dependent manner. Among the three hydrophobic 4-nitroanilides of alanine, valine and leucine, the latter displays the largest overall activation (increase in kcat/Km). Large enhancements in affinity (1/Ki) upon Ca2+ binding have been observed for inhibitors with flexible (leucine-like) residues at their N-termini and smaller enhancements for inhibitors with rigid (phenylalanine-like) residues. [ABSTRACT FROM AUTHOR]

The patch clamp technique has been used to investigate ion permeation and Ca(2+)-dependent gating of a voltage-sensitive Ca2+ release channel in the vacuolar membrane of sugar beet tap roots. Reversal potential measurements in bi-ionic conditions revealed a sequence for permeability ratios of Ca2+ approximately Sr2+ approximately Ba2+ > Mg2+ >> K+ which is inversely related to the size of the unitary conductances K+ >> Mg2+ approximately Ba2+ > Sr2+ approximately Ca2+, suggesting that ion movement is not independent. In the presence of Ca2+, the unitary K+ current is reduced in a concentration- and voltage-dependent manner by Ca2+ binding at a high affinity site (K0.5 = 0.29 mM at 0 mV) which is located 9% along the electric field of the membrane from the vacuolar side. Comparison of reversal potentials obtained under strictly bi-ionic conditions with those obtained in the presence of mixtures of the two ions indicates that the channel forms a multi-ion pore. Lumenal Ca2+ also has an effect on voltage-dependent channel gating. Stepwise increases of vacuolar Ca2+ from micromolar to millimolar concentrations resulted in a dramatic increase in channel openings over the physiological voltage range via a shift in threshold for channel activation to less negative membrane potentials. The steepness of the concentration dependence of channel activation by Ca2+ at -41 mV predicts that two Ca2+ ions need to bind to open the gate. The implications of the results for ion permeation and channel gating are discussed. [ABSTRACT FROM AUTHOR]

Ca2+-modulated rod outer segment membrane guanylate cyclase (ROS-GC1) has been cloned and reconstituted to show that it is regulated by two processes: one inhibitory, the other stimulatory. The inhibitory process is consistent with its linkage to phototransduction; the physiology of the stimulatory process is probably linked to neuronal transmission. In both regulatory processes, calcium modulation of the cyclase takes place through the calcium binding proteins; guanylate cyclase activating proteins (GCAP1 and GCAP2) in the case of the phototransduction process and calcium-dependent GCAP (CD-GCAP) in the case of the stimulatory process. The cyclase domains involved in the two processes are located at two different sites on the ROS-GC1 intracellular region. The GCAP1-modulated domain resides within the aa 447-730 segment of ROS-GC1 and the CD-GCAP-modulated domain resides within the aa 731-1054 segment. In the present study the GCAP2-dependent Ca2+ modulation of the cyclase activity has been reconstituted using recombinant forms of GCAP2 and ROS-GC1, and its mutants. The results indicate that consistent to phototransduction, GCAP2 at low Ca2+ concentration (10 nM) maximally stimulates the cyclase activity of the wild-type and its mutants: ext- (deleted aa 8-408); kin- (deleted aa 447-730) and hybrid consisting of the ext, transmembrane and kin domains of ANF-RGC and the C-terminal domain, aa 731-1054, of ROS-GC1. In all cases, it inhibits the cyclase activity with an IC50 of about 140 nM. A previous study has shown that under identical conditions the kin- and the hybrid mutant are at best only minimally stimulated. Thus, the GCAP1 and GCAP2 signal transduction mechanisms are different, occurring through different modules of ROS-GC1. These findings also demonstrate that the intracellular region of ROS-GC1 is composed of multiple modules, each designed to mediate a particular calcium-specific signalling pathway. [ABSTRACT FROM AUTHOR]

In this study, we aim to explore the anti-tumor effect of liraglutide (Lira), an anti-diabetic medicine, on pancreatic cancer cell PANC-1 co-cultured with or without pancreatic stellate cells (PSCs). The chemical count kit-8 and Annexin V-FITC apoptosis detection were conducted to investigate the effect of Lira on cell viability and proliferation of PANC-1 with or without PSCs co-culture. Then, the wound healing and transwell experiments were performed to explore the influence of Lira on PANC-1 cells' migration and invasion capabilities. To identify the potential action mechanism of Lira on PANC-1, the expression of E-cadherin and N-cadherin and the intracellular calcium content in PANC-1, after Lira administration, were detected. The results indicated that Lira in 100 and 1,000 nmol/L, effectively decreased the cell viability and dose-dependently promoted cell apoptosis of PANC-1 co-cultured with or without PSCs. Lira significantly reduced the migration and invasion of PANC-1 and also reduced the inducing effect of PSCs to PANC-1. Lira effectively induced the expression of E-cadherin and suppressed the expression of N-cadherin with a dose-dependent manner. Otherwise, Lira significantly reduced the abnormal high content of calcium in PANC-1 and also weakened the elevation of calcium in PANC-1 induced by cell-cell interaction. The current study firstly indicated that Lira suppressed the cell proliferation, migration and invasion of PANC-1 with or without PSCs co-culture. This effect was partially due to the calcium modulation of Lira and its influence on Ca 2+ -binding proteins, such as E-cadherin and N-cadherin.