11 results on '"V. Ghiaroni"'
Search Results
2. Effect of ciguatoxin 3C on voltage-gated Na+ and K+ currents in mouse taste cells.
- Author
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Ghiaroni V, Fuwa H, Inoue M, Sasaki M, Miyazaki K, Hirama M, Yasumoto T, Rossini GP, Scalera G, and Bigiani A
- Subjects
- Animals, Ciguatoxins chemistry, Ethers, Cyclic chemistry, Ethers, Cyclic pharmacology, Mice, Organ Culture Techniques, Patch-Clamp Techniques, Polycyclic Compounds chemistry, Polycyclic Compounds pharmacology, Potassium Channels, Voltage-Gated drug effects, Taste drug effects, Taste physiology, Ciguatoxins pharmacology, Ion Channel Gating drug effects, Sodium Channels drug effects, Taste Buds cytology, Taste Buds drug effects
- Abstract
The marine dinoflagellate Gambierdiscus toxicus produces highly lipophilic, polycyclic ether toxins that cause a seafood poisoning called ciguatera. Ciguatoxins (CTXs) and gambierol represent the two major causative agents of ciguatera intoxication, which include taste alterations (dysgeusiae). However, information on the mode of action of ciguatera toxins in taste cells is scarce. Here, we have studied the effect of synthetic CTX3C (a CTX congener) on mouse taste cells. By using the patch-clamp technique to monitor membrane ion currents, we found that CTX3C markedly affected the operation of voltage-gated Na(+) channels but was ineffective on voltage-gated K(+) channels. This result was the exact opposite of what we obtained earlier with gambierol, which inhibits K(+) channels but not Na(+) channels. Thus, CTXs and gambierol affect with high potency the operation of separate classes of voltage-gated ion channels in taste cells. Our data suggest that taste disturbances reported in ciguatera poisoning might be due to the ability of ciguatera toxins to interfere with ion channels in taste buds.
- Published
- 2006
- Full Text
- View/download PDF
3. A dynamic population of excitable cells: the taste receptor cells.
- Author
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Ghiaroni V, Fieni F, Silvestri F, Pietra P, and Bigiani A
- Subjects
- Action Potentials physiology, Aging physiology, Animals, Cell Differentiation drug effects, Cell Membrane drug effects, Chloride Channels drug effects, Chloride Channels physiology, In Vitro Techniques, Ion Channels drug effects, Mice, Patch-Clamp Techniques, Potassium Channel Blockers pharmacology, Potassium Channels drug effects, Potassium Channels physiology, Signal Transduction drug effects, Signal Transduction physiology, Sodium Channels physiology, Taste Buds cytology, Taste Buds drug effects, Cell Differentiation physiology, Cell Membrane physiology, Ion Channels physiology, Taste Buds physiology
- Abstract
Taste receptor cells (TRCs) represent an unique opportunity to study a dynamic population of excitable cells that undergoes two basic neurobiological processes: postnatal development and cell turnover. We have begun to investigate the functional properties of TRCs and how they mature over time by applying the patch-clamp technique to single cell in taste buds isolated from mouse vallate papilla during postnatal development. We have focussed our attention on a well-defined functional group of taste cells, called Na/OUT cells, and on their voltage-gated K+, and Cl- currents (I(K) and I(Cl), respectively). As in neurons, I(K) and I(Cl) underlie action potential waveform and firing properties in these cells. By analyzing the relative occurrence of I(K) and I(Cl) among cells, we found that in adult mice three different electrophysiological phenotypes of Na/OUT cells could be detected: cells with only I(K) (K cells); cells with both I(K) and I(Cl) (K + Cl cells); and cells with I(Cl) (Cl cells). On the contrary, at early developmental stages (2-4 postnatal day, PD) there were no Cl cells, which appeared at PD 8. The analysis of the changes in current amplitude (which continuously increased in developing cells) during postnatal development suggested that Cl cells and K + Cl cells likely represented a single functional line different from K cells. In addition, electrophysiological data were consistent with the interpretation that Cl cells derived from some K + Cl cells by suppression of I(K). The dynamics of the expression of I(K) and I(Cl) during postnatal development likely reflects a mechanism that could also operate during turnover.
- Published
- 2005
4. Inhibition of voltage-gated potassium currents by gambierol in mouse taste cells.
- Author
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Ghiaroni V, Sasaki M, Fuwa H, Rossini GP, Scalera G, Yasumoto T, Pietra P, and Bigiani A
- Subjects
- Animals, Cells, Cultured, Mice, Patch-Clamp Techniques, Sodium Channels metabolism, Taste Buds cytology, Taste Buds metabolism, Action Potentials drug effects, Ciguatoxins toxicity, Ethers, Cyclic toxicity, Polycyclic Compounds toxicity, Potassium Channels, Voltage-Gated antagonists & inhibitors, Taste Buds drug effects
- Abstract
Ciguatera is a food poisoning caused by toxins of Gambierdiscus toxicus, a marine dinoflagellate. The neurological features of this intoxication include sensory abnormalities, such as paraesthesia, heightened nociperception, and also taste alterations. Here, we have evaluated the effect of gambierol, one of the possible ciguatera toxins, on the voltage-gated ion currents in taste cells. Taste cells are excitable cells endowed with voltage-gated Na+, K+, and Cl- currents (I(Na), I(K), and I(Cl), respectively). By applying the patch-clamp technique to single cells in isolated taste buds obtained from the mouse vallate papilla, we have recorded such currents and determined the effect of bath-applied gambierol. We found that this toxin markedly inhibited I(K) in the nanomolar range (IC50 of 1.8 nM), whereas it showed no significant effect on I(Na) or I(Cl) even at high concentration (1 microM). The block of I(K) was irreversible even after a 50-min wash. In addition to affecting the current amplitude, we found that gambierol significantly altered both the activation and inactivation processes of I(K). In conclusion, unlike other toxins involved in ciguatera, such as ciguatoxins, which affect the functioning of voltage-gated sodium channels, the preferred molecular target of gambierol is the voltage-gated potassium channel, at least in taste cells. Voltage-gated potassium currents play an important role in the generation of the firing pattern during chemotransduction. Thus, gambierol may alter action potential discharge in taste cells and this could be associated with the taste alterations reported in the clinical literature.
- Published
- 2005
- Full Text
- View/download PDF
5. Functional correlates of somatostatin receptor 2 overexpression in the retina of mice with genetic deletion of somatostatin receptor 1.
- Author
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Bigiani A, Petrucci C, Ghiaroni V, Dal Monte M, Cozzi A, Kreienkamp HJ, Richter D, and Bagnoli P
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- Animals, Female, Gene Expression Regulation drug effects, Gene Expression Regulation physiology, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Octreotide pharmacology, Receptors, Somatostatin agonists, Receptors, Somatostatin biosynthesis, Retina drug effects, Gene Deletion, Receptors, Somatostatin genetics, Receptors, Somatostatin physiology, Retina metabolism
- Abstract
Somatostatin-14 (SRIF) and its receptors (sst(1-5)) are found in the mammalian retina. However, scarce information is available on the role of the somatostatinergic system in retinal physiology. We have recently used gene-knockout technology to gain insights into the function of sst(1) and sst(2) receptors in the mouse retina. The sst(1) receptor localizes to SRIF-containing amacrine cells, whereas the sst(2) receptor localizes to several retinal cell populations including rod bipolar cells (RBCs). Molecular data indicate that, in retinas with deletion of the sst(1) receptor (sst(1) KO), sst(2) receptors become overexpressed in concomitance with an increased level of retinal SRIF. To test whether this up-regulation of sst(2) receptors correlates with altered sst(2) receptor physiology, we studied the effect of sst(2) receptor activation on potassium current (I(K)) in isolated RBCs and glutamate release in retina explants. Both I(K) and glutamate release are known to be negatively modulated by sst(2) receptors in the mammalian retina. We used octreotide, a SRIF analogue, to activate selectively sst(2) receptors. Patch-clamp recordings from isolated RBCs indicated that the sst(2) receptor-mediated inhibition of I(K) was significantly larger in sst(1) KO than in control retinas. In addition, HPLC measurements of glutamate release in sst(1) KO retinal explants demonstrated that the sst(2) receptor-mediated inhibition of K(+)-evoked glutamate release was also significantly larger than in control retinas. As a whole, these findings indicate that the overexpression of sst(2) receptors in sst(1) KO retinas can be correlated to an enhanced function of sst(2) receptors. The level of expression of sst(2) receptors may therefore represent a key step in the regulation of sst(2) receptor-mediated responses, at least in the retina.
- Published
- 2004
- Full Text
- View/download PDF
6. Adrenocorticotropin reverses hemorrhagic shock in anesthetized rats through the rapid activation of a vagal anti-inflammatory pathway.
- Author
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Guarini S, Cainazzo MM, Giuliani D, Mioni C, Altavilla D, Marini H, Bigiani A, Ghiaroni V, Passaniti M, Leone S, Bazzani C, Caputi AP, Squadrito F, and Bertolini A
- Subjects
- Acute Disease, Animals, Atropine pharmacology, Chlorisondamine therapeutic use, Electrophoretic Mobility Shift Assay, Female, I-kappa B Proteins metabolism, Liver metabolism, Male, NF-kappa B metabolism, RNA, Messenger metabolism, Rats, Rats, Wistar, Receptors, Nicotinic drug effects, Reverse Transcriptase Polymerase Chain Reaction, Tumor Necrosis Factor-alpha analysis, Tumor Necrosis Factor-alpha genetics, Vagus Nerve drug effects, Cosyntropin therapeutic use, Shock, Hemorrhagic drug therapy, Shock, Hemorrhagic physiopathology, Vagus Nerve physiopathology
- Abstract
Objective: Several melanocortin peptides have a prompt and sustained resuscitating effect in conditions of hemorrhagic shock. The transcription nuclear factor kappaB (NF-kappaB) triggers a potentially lethal systemic inflammatory response, with marked production of tumor necrosis factor-alpha (TNF-alpha), in hemorrhagic shock. Here we investigated whether the hemorrhagic shock reversal produced by the melanocortin ACTH-(1-24) (adrenocorticotropin) depends on the activation of the recently recognized, vagus nerve-mediated, brain "cholinergic anti-inflammatory pathway"., Methods and Results: Anesthetized rats were stepwise bled until mean arterial pressure (MAP) stabilized at 20-25 mm Hg. The severe hypovolemia was incompatible with survival, and all saline-treated animals died within 30 min. In rats intravenously (i.v.) treated with ACTH-(1-24), neural efferent activity along vagus nerve (monitored by means of a standard system for extracellular recordings) was markedly increased, and the restoration of cardiovascular and respiratory functions was associated with blunted NF-kappaB activity and with decreased TNF-alpha mRNA liver content and TNF-alpha plasma levels. Bilateral cervical vagotomy, pretreatment with the melanocortin MC(4) receptor antagonist HS014, atropine sulfate or chlorisondamine, but not with atropine methylbromide, prevented the life-saving effect of ACTH-(1-24) and the associated effects on NF-kappaB activity and TNF-alpha levels. HS014 and atropine sulfate prevented, too, the ACTH-(1-24)-induced increase in neural efferent vagal activity, and accelerated the evolution of shock in saline-treated rats., Conclusions: The present data show, for the first time, that the melanocortin ACTH-(1-24) suppresses the NF-kappaB-dependent systemic inflammatory response triggered by hemorrhage, and reverses shock condition, by brain activation (in real-time) of the "cholinergic anti-inflammatory pathway", this pathway seeming to be melanocortin-dependent., (Copyright 2004 European Society of Cardiology)
- Published
- 2004
- Full Text
- View/download PDF
7. Electrophysiological heterogeneity in a functional subset of mouse taste cells during postnatal development.
- Author
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Ghiaroni V, Fieni F, Pietra P, and Bigiani A
- Subjects
- Animals, Chlorides metabolism, Electrophysiology, Ion Channel Gating, Mice, Patch-Clamp Techniques, Potassium metabolism, Sodium metabolism, Taste Buds cytology, Aging physiology, Taste physiology, Taste Buds growth & development, Taste Buds metabolism
- Abstract
Taste cells in adult mammals are functionally heterogeneous as to the expression of ion channels. How these adult phenotypes are established during postnatal development, however, is not yet clear. We have addressed this issue by studying voltage-gated K(+) and Cl(-) currents (I(K) and I(Cl), respectively) in developing taste cells of the mouse vallate papilla. I(K) and I(Cl) underlie action potential waveform and firing properties, and play an important role in taste transduction. By using the patch clamp technique, we analyzed these currents in a specific group of cells, called Na/OUT cells and thought to be sensory. In adult mice, three different electrophysiological phenotypes of Na/OUT cells could be detected: cells with I(K) (K cells); cells with both I(K) and I(Cl) (K+Cl cells); and cells with I(Cl) (Cl cells). In contrast, at early developmental stages (2-4 postnatal days, PD) there were no Cl cells, which appeared at PD 8. Our findings indicate a mechanism that contributes to building-up the functional heterogeneity of mammalian taste cells during the postnatal development.
- Published
- 2003
- Full Text
- View/download PDF
8. Channels as taste receptors in vertebrates.
- Author
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Bigiani A, Ghiaroni V, and Fieni F
- Subjects
- Animals, Anura, Epithelial Sodium Channels, Humans, Membrane Potentials physiology, Necturus, Potassium Channels, Voltage-Gated physiology, Signal Transduction physiology, Sodium Channels physiology, Stimulation, Chemical, Ion Channels physiology, Sensory Receptor Cells physiology, Taste Buds physiology
- Abstract
Taste reception is fundamental for proper selection of food and beverages. Chemicals detected as taste stimuli by vertebrates include a large variety of substances, ranging from inorganic ions (e.g., Na(+), H(+)) to more complex molecules (e.g., sucrose, amino acids, alkaloids). Specialized epithelial cells, called taste receptor cells (TRCs), express specific membrane proteins that function as receptors for taste stimuli. Classical view of the early events in chemical detection was based on the assumption that taste substances bind to membrane receptors in TRCs without permeating the tissue. Although this model is still valid for some chemicals, such as sucrose, it does not hold for small ions, such as Na(+), that actually diffuse inside the taste tissue through ion channels. Electrophysiological, pharmacological, biochemical, and molecular biological studies have provided evidence that indeed TRCs use ion channels to reveal the presence of certain substances in foodstuff. In this review, we focus on the functional and molecular properties of ion channels that serve as receptors in taste transduction.
- Published
- 2003
- Full Text
- View/download PDF
9. Apical and basal neurones isolated from the mouse vomeronasal organ differ for voltage-dependent currents.
- Author
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Fieni F, Ghiaroni V, Tirindelli R, Pietra P, and Bigiani A
- Subjects
- Action Potentials physiology, Animals, Calcium Channels physiology, Electrophysiology, Female, In Situ Hybridization, In Vitro Techniques, Ion Channel Gating physiology, Male, Membrane Potentials physiology, Mice, Patch-Clamp Techniques, Potassium Channels physiology, Sodium Channels physiology, Tetrodotoxin pharmacology, Vomeronasal Organ innervation, Ion Channels physiology, Neurons physiology, Vomeronasal Organ physiology
- Abstract
The mammalian vomeronasal organ (VNO) contains specialized neurones that transduce the chemical information related to pheromones into discharge of action potentials to the brain. Molecular and biochemical studies have shown that specific components of the pheromonal transduction systems are segregated into two distinct subsets of vomeronasal neurones: apical neurones and basal neurones. However, it is still unknown whether these neuronal subsets also differ in other functional characteristics, such as their membrane properties. We addressed this issue by studying the electrophysiological properties of vomeronasal neurones isolated from mouse VNO. We used the patch-clamp technique to examine both the passive membrane properties and the voltage-gated Na+, K+ and Ca2+ currents. Apical neurones were distinguished from basal ones by the length of their dendrites and by their distinct immunoreactivity for the putative pheromone receptor V2R2. The analysis of passive properties revealed that there were no significant differences between the two neuronal subsets. Also, apical neurones were similar to basal neurones in their biophysical and pharmacological properties of voltage-gated Na+ and K+ currents. However, we found that the density of Na+ currents was about 2-3 times greater in apical neurones than in basal neurones. Consistently, in situ hybridization analysis revealed a higher expression of the Na+ channel subtype III in apical neurones than in basal ones. In contrast, basal neurones were endowed with Ca2+ currents (T-type) of greater magnitude than apical neurones. Our findings indicate that apical and basal neurones in the VNO exhibit distinct electrical properties. This might have a profound effect on the sensory processes occurring in the VNO during pheromone detection.
- Published
- 2003
- Full Text
- View/download PDF
10. Ion conductances in supporting cells isolated from the mouse vomeronasal organ.
- Author
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Ghiaroni V, Fieni F, Tirindelli R, Pietra P, and Bigiani A
- Subjects
- Animals, Male, Membrane Potentials physiology, Mice, Mice, Inbred C57BL, Neurons, Afferent physiology, Patch-Clamp Techniques, Potassium metabolism, Potassium Channels, Voltage-Gated physiology, Signal Transduction physiology, Sodium metabolism, Sodium Channels physiology, Vomeronasal Organ innervation, Neuroglia physiology, Vomeronasal Organ cytology, Vomeronasal Organ physiology
- Abstract
The vomeronasal organ (VNO) is a chemosensory structure involved in the detection of pheromones in most mammals. The VNO sensory epithelium contains both neurons and supporting cells. Data suggest that vomeronasal neurons represent the pheromonal transduction sites, whereas scarce information is available on the functional properties of supporting cells. To begin to understand their role in VNO physiology, we have characterized with patch-clamp recording techniques the electrophysiological properties of supporting cells isolated from the neuroepithelium of the mouse VNO. Supporting cells were distinguished from neurons by their typical morphology and by the lack of immunoreactivity for Ggamma8 and OMP, two specific markers for vomeronasal neurons. Unlike glial cells in other tissues, VNO supporting cells exhibited a depolarized resting potential (about -29 mV). A Goldman-Hodgkin-Katz analysis for resting ion permeabilities revealed indeed an unique ratio of P(K):P(Na):P(Cl) = 1:0.23:1.4. Supporting cells also possessed voltage-dependent K(+) and Na(+) conductances that differed significantly in their biophysical and pharmacological properties from those expressed by VNO neurons. Thus glial membranes in the VNO can sustain significant fluxes of K(+) and Na(+), as well as Cl(-). This functional property might allow supporting cells to mop-up and redistribute the excess of KCl and NaCl that often occurs in certain pheromone-delivering fluids, like urine, and that could blunt the sensitivity of VNO neurons to pheromones. Therefore vomeronasal supporting cells could affect chemosensory transduction in the VNO by regulating the ionic strength of the pheromone-containing medium.
- Published
- 2003
- Full Text
- View/download PDF
11. Postnatal development of membrane excitability in taste cells of the mouse vallate papilla.
- Author
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Bigiani A, Cristiani R, Fieni F, Ghiaroni V, Bagnoli P, and Pietra P
- Subjects
- 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid pharmacology, Action Potentials drug effects, Action Potentials physiology, Animals, Cell Count, Cell Membrane drug effects, Chloride Channels metabolism, Chlorides metabolism, In Vitro Techniques, Membrane Potentials drug effects, Membrane Potentials physiology, Mice, Mice, Inbred C57BL, Patch-Clamp Techniques, Potassium metabolism, Potassium Channel Blockers pharmacology, Potassium Channels, Voltage-Gated antagonists & inhibitors, Potassium Channels, Voltage-Gated metabolism, Sodium metabolism, Sodium Channel Blockers, Sodium Channels metabolism, Taste Buds cytology, Taste Buds drug effects, Tetrodotoxin pharmacology, Transducin metabolism, Aging metabolism, Cell Membrane physiology, Taste Buds growth & development, Taste Buds metabolism
- Abstract
The mammalian peripheral taste system undergoes functional changes during postnatal development. These changes could reflect age-dependent alterations in the membrane properties of taste cells, which use a vast array of ion channels for transduction mechanisms. Yet, scarce information is available on the membrane events in developing taste cells. We have addressed this issue by studying voltage-dependent Na+, K+, and Cl- currents (I(Na), I(K), and I(Cl), respectively) in a subset of taste cells (the so-called "Na/OUT" cells, which are electrically excitable and thought to be sensory) from mouse vallate papilla. Voltage-dependent currents play a key role during taste transduction, especially in the generation of action potentials. Patch-clamp recordings revealed that I(Na), I(K), and I(Cl) were expressed early in postnatal development. However, only I(K) and I(Cl) densities increased significantly in developing Na/OUT cells. Consistent with the rise of I(K) density, we found that action potential waveform changed markedly, with an increased speed of repolarization that was accompanied by an enhanced capability of repetitive firing. In addition to membrane excitability changes in putative sensory cells, we observed a concomitant increase in the occurrence of glia-like taste cells (the so called "leaky" cells) among patched cells. Leaky cells are likely involved in dissipating the increase of extracellular K+ during action potential discharge in chemosensory cells. Thus, developing taste cells of the mouse vallate papilla undergo a significant electrophysiological maturation and diversification. These functional changes may have a profound impact on the transduction capabilities of taste buds during development.
- Published
- 2002
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