PubMed Health⌕ Search

Biomedical subjects

L Cazin

Publications and source records attributed to L Cazin.

At least 19 recordsLinked to original sources
PubMed1998

Adenosine potentiates the delayed-rectifier potassium conductance but has no effect on the hyperpolarization-activated Ih current in frog melanotrophs.

The effects of adenosine on the voltage-sensitive delayed-rectifier K+ (IK) currents and hyperpolarization-activated cationic inward current (Ih) were studied in cultured frog melanotrophs using the whole-cell configuration of the patch-clamp technique. The A1 receptor agonist R-N6-phenylisopropyl-adenosine (R-PIA; 50 microM) reversibly increased IK. Perfusion of dibutyryl-cAMP (1 mM) in the external solution did not modify the R-PIA-induced enhancement of IK. Pretreatment of melanotrophs with pertussis toxin (1 microg/ml; 12 h) totally abolished the R-PIA-evoked response. Application of hyperpolarizing voltage pulses from -60 to -120 mV to melanotrophs induced a two-component inward current corresponding to an Ih-like conductance. This conductance was characterized by a high K+ selectivity and a low Na+ permeability and was resistant to tetrodotoxin (1 microM). R-PIA had no effect on Ih. The present study demonstrates that in frog melanotrophs adenosine inhibits the electrical activity by activating IK through an A1 receptor subtype coupled to a pertussis toxin-sensitive pathway independent of the cAMP/PKA system. This study also demonstrates the existence of a Ih conductance in frog melanotrophs which is not modulated by A1 receptors.

Adenosine↗
PubMed1998

Gramicidin-perforated patch revealed depolarizing effect of GABA in cultured frog melanotrophs.

1. In frog pituitary melanotrophs, GABA induces a transient stimulation followed by prolonged inhibition of hormone secretion. This biphasic effect is inconsistent with the elevation of cytosolic calcium and the inhibition of electrical activity also provoked by GABA in single melanotrophs. In the present study, standard patch-clamp configurations and gramicidin-perforated patches were used to investigate the physiological GABAA receptor-mediated response and intracellular chloride concentration ([Cl-]i) in cultured frog melanotrophs. 2. In the gramicidin-perforated patch configuration, 1 microM GABA caused a depolarization associated with an action potential discharge and a slight fall of membrane resistance. In contrast, at a higher concentration (10 microM) GABA elicited a depolarization accompanied by a transient volley of action potentials, followed by a sustained inhibitory plateau and a marked fall of membrane resistance. Isoguvacine mimicked the GABA-evoked responses, indicating a mediation by GABAA receptors. 3. In gramicidin-perforated cells, the depolarizing excitatory effect of 1 microM GABA was converted into a depolarizing inhibitory action when 0.4 microM allopregnanolone was added to the bath solution. 4. After gaining the whole-cell configuration, the amplitude and/or direction of the GABA-evoked current (IGABA) rapidly changed before stabilizing. After stabilization, the reversal potential of IGABA followed the values predicted by the Nernst equation for chloride ions when [Cl-]i was varied. 5. In gramicidin-perforated cells, the steady-state I-V relationships of 10 microM GABA- or isoguvacine-evoked currents yielded reversal potentials of -37.5 +/- 1.6 (n = 17) and -38.6 +/- 2.0 mV (n = 8), respectively. These values were close to those obtained by using a voltage-ramp protocol in the presence of Na+, K+ and Ca2+ channel blockers. The current evoked by 1 microM GABA also reversed at these potentials. 6. We conclude that, in frog pituitary melanotrophs, chloride is the exclusive charge carrier of IGABA. In intact cells, the reversal potential of IGABA is positive to the resting potential because of a relatively high [Cl-]i (26.5 mM). Under these conditions, GABA induces a chloride efflux responsible for a depolarization triggering action potentials. However, GABA at a high concentration or in the presence of the potentiating steroid allopregnanolone exerts a concomitant shunting effect leading to a rapid inhibition of the spontaneous firing.

Animals↗
PubMed1998

Sigma ligands stimulate the electrical activity of frog pituitary melanotrope cells through a G-protein-dependent inhibition of potassium conductances.

We have investigated the effects of sigma ligands [1,3-di(2-tolyl)guanidine (DTG) and (+)-pentazocine] on the electrical activity of cultured frog pituitary melanotrope cells by using the patch-clamp technique. DTG and (+)-pentazocine (10 microM each) induced a reversible depolarization associated with an increase in membrane resistance and action potential firing. In voltage-clamp experiments, DTG and (+)-pentazocine elicited inward currents whose intensity augmented with membrane depolarization. The currents vanished or reversed between -90 and -100 mV, at values close to the K+ equilibrium potential (E(K)+ = -102 mV). DTG (2-500 microM) and (+)-pentazocine (0.2-200 microM) reduced the outward delayed rectifier K+ current [IK (V)] in a dose-dependent manner with EC50 of 64 and 37 microM, respectively. In contrast, naloxone (50 microM) and pirenzepine (10 microM) did not affect the sigma ligand-induced inhibition of IK (V). Addition of guanosine-5'-O-(3-thiophosphate) in the pipette solution irreversibly sustained the DTG-induced current whereas guanosine-5'-O-(2-thiodiphosphate) virtually suppressed the response. Cholera toxin-pretreatment (1 microgram/ml; 18 hr) abolished the inward current and the inhibition of IK (V) induced by sigma ligands. In contrast, pretreatment with pertussis toxin (1 microgram/ml; 18 hr) had no effect. Taken together, these data indicate that DTG and (+)-pentazocine activate the electrical activity of cultured frog melanotrope cells by reducing both a tonic K+ current and a voltage-dependent [IK (V)] K+ conductance through the activation of a cholera toxin-sensitive G-protein.

Action Potentials↗
PubMed1997

Multiple modulatory effects of the neuroactive steroid pregnanolone on GABAA receptor in frog pituitary melanotrophs.

1. The effects of the neuroactive steroid pregnanolone (5 beta-pregnan-3 alpha-ol-20-one) on the electrical response to GABA were investigated in cultured frog pituitary melanotrophs using the patch-clamp technique. 2. Low concentrations of pregnanolone (0.01-1 microM) in the extracellular solution enhanced the current evoked by submaximal concentrations of GABAA receptor agonists and prolonged the GABA-induced inhibition of the spontaneous action potentials in a dose-dependent manner. 3. Pregnanolone augmented the opening probability of the single GABA-activated channels but did not modify the conductance levels. 4. Pregnanolone (1 microM) shifted the GABA dose-response curve towards the low GABA concentrations, reducing the EC50 from 4.2 to 1.8 microM. 5. Internal cell dialysis with pregnanolone (1 or 10 microM) did not alter the GABA-evoked current. 6. Pregnanolone accelerated the desensitization of both the current and conductance increases caused by GABA. 7. High concentrations of pregnanolone (30 microM) markedly and reversibly diminished the current evoked by 10 microM GABA. 8. At high concentrations (10-30 microM), pregnanolone induced an outward current which reversed at the chloride equilibrium potential. 9. It is concluded that, in frog pituitary melanotrophs, pregnanolone exerts a dual inverse modulation and a direct activation of the GABAA receptor-channel depending on the concentrations of both GABA and steroid. Pregnanolone acts on an extracellular site on the GABAA receptor inducing conformational changes of the receptor-channel complex, resulting in a desensitized less-conducting state.

Action Potentials↗
PubMed1997

Electrophysiological effects of various neuroactive steroids on the GABA(A) receptor in pituitary melanotrope cells.

The action of steroids on the bioelectrical response to gamma-aminobutyric acid (GABA) has never been studied in pituitary cells. In the present study, we have thus investigated the effects of a series of neuroactive steroids on the GABA-activated current in frog melanotrope cells in primary culture, using the patch-clamp technique in the whole-cell configuration. Bath perfusion of 3alpha-isomers of pregnanolone or tetrahydrodeoxycorticosterone (1 microM) significantly enhanced the current evoked by short pulses of GABA (3 microM) and accelerated its desensitization. In contrast, the 3beta-isomers (30 microM) had no effect on the GABA-activated current. Addition to the bath solution of dehydroepiandrosterone or dehydroepiandrosterone sulfate (10 microM) inhibited the GABA-activated current without modifying its kinetics while pregnenolone sulfate (10 microM) both inhibited the GABA-activated current and accelerated its decay rate. The effects of pregnane steroids were not impaired by the central-type benzodiazepine receptor antagonist flumazenil (10 microM). In conclusion, the present study reveals that neuroactive steroids may exert multiple modulatory activities on the GABA(A) receptor borne by melanotrope cells. The effect of steroids on the current evoked by GABA is rapid, reversible, stereospecific and not mediated through the benzodiazepine binding site of the GABA(A) receptor.

Animals↗
PubMed1997

[Permanent percutaneous electric connection. General principles].

The Swedes for more than twenty years, and the Germans for over five years have been able to maintain inert or active prostheses with permanent percutaneous connections, thanks to the dependable and proven material and techniques of extraoral implants. The significant improvement extra-oral implants have brought about is not only in a new therapeutic approach to the treatment of important facial defects or transmission deafness; it is also because for some twenty old years now, the few millimeter wide cylinders of Titanium, the < > affixed on the implants, have crossed the cutaneous barrier for extended periods without complications. The percutaneous abutment thus creates a permanent communication between the interior and the exterior of the organism. If the abutment, instead of simply carrying a Maxillo-Facial Prosthesis or an auditive prosthesis, is modified by placing an electric conductor inside it, the simple "percutaneous peg" will turn out to be, in a way, a "percutaneous electric plug". By adapting classic "mechanical" abutments and implants, authors have created a Permanent Percutaneous Electric Connection (PPEC) which has been successfully experimented on rabbits to record EEG. Clinical applications on humans would make it possible either to receive "bio-electrical information" coming from within the organism, or to send electrical energy into the organism. This last application opens vast perspectives of improvement both in diagnosis and therapy in many fields.

Humans↗
PubMed1996

Adenosine inhibits L- and N-type calcium channels in pituitary melanotrophs. Evidence for the involvement of a G protein in calcium channel gating.

It has been previously demonstrated that activation of A1 adenosine receptors in frog melanotrophs causes inhibition of spontaneous action potential discharges and alpha-melanocyte-stimulating hormone secretion. In the present study, we have investigated the effect of adenosine on high-voltage-activated (HVA) calcium currents in cultured melanotrophs, using the whole-cell variant of the patch-clamp technique with barium as a charge carrier. Adenosine and the specific A1 adenosine receptor agonist R-PIA (50 microM each) produced a decrease of the amplitude of the barium current, while the selective A2 adenosine receptor agonist CGS 21680 did not affect the current. The inhibitory effect of R-PIA was observed throughout the activation range of the current, with stronger responses at more positive potentials. R-PIA inhibited both the L- and N-type components of the current, the effect on the N-component being two-fold higher than on the L-component. The inhibitory effect of R-PIA was rendered irreversible by addition of GTP gamma S (100 microM) to the intracellular solution. Pre-treatment of the cells with pertussis toxin (1 microgram/ml; 12 h) totally abolished the effect of R-PIA on the HVA calcium channels. Conversely, addition of a high concentration of cAMP (100 microM) together with the phosphodiesterase inhibitor IBMX (100 microM) to the intracellular solution did not modify the effect of R-PIA on the current. It is concluded that, in frog melanotrophs, adenosine induces inhibition of L- and N-calcium currents and that this effect is mediated by a pertussis toxin-sensitive G protein. Our data also indicate that the inhibitory effect of adenosine on the calcium currents is not mediated by inhibition of adenylyl cyclase.

1-Methyl-3-isobutylxanthine↗
PubMed1995

Osmo- and mechanosensitivity of the transient outward K+ current in a mammalian neuronal cell line.

1. The transient outward current in NG108-15 cells was investigated with the whole-cell patch-clamp technique. The current was inhibited by external 4-aminopyridine or tetra-ethylammonium. The reversal potential shifted rightward with increased external K+ concentrations. 2. Current inactivation was markedly accelerated in hyperosmotic media (+30 mosmol l-1) and after nearby ejection of isosmotic solution with maximal acceleration occurring after 15-20 s and full recovery within 2-4 min, thus demonstrating an osmo- and mechanosensitivity of this current. Voltage-dependent Na+ and Ca2+ currents were unaffected. 3. Hyperosmotic solution shifted the voltage dependence of inactivation leftward. Inactivation was sensitive to reducing and oxidizing intracellular conditions. Reduction blocked the acceleration of current inactivation induced by hyperosmotic media, while oxidation did not hamper the response. 4. Action potentials had a decreased amplitude and a slower repolarization after hyperosmotic ejections. 5. It is concluded that the transient K+ current is osmo- and mechanosensitive, thus providing a mechanism for extracellular osmolarity to modulate neuronal excitability. The response appeared to be mediated through a changed sensitivity of the inactivating principle to the membrane electric field and was dependent on the redox state of the cell.

4-Aminopyridine↗
PubMed1995

A-type potassium current modulated by A1 adenosine receptor in frog melanotrophs.

1. Transient outward current was recorded in cultured frog melanotrophs with the whole-cell configuration of the patch-clamp technique. The ionic dependence, kinetics and pharmacological properties of the current were studied. The effects of the A1 adenosine receptor agonist R-N6-phenylisopropyl-adenosine (R-PIA) on this current were also investigated. 2. In tetrodotoxin- and cobalt-containing solution, depolarization from -120 mV elicited both transient and delayed outward currents. Pulses from -60 mV activated only a sustained late current. 3. 4-Aminopyridine (4 mM) reduced the transient outward current much more than the delayed outward current. In contrast, tetraethylammonium (10-20 mM) selectively reduced the delayed current. 4. Tail current measurements showed a positive shift in the reversal potential when external K+ concentration was increased, indicating that K+ was the predominant charge carrier. 5. Steady-state inactivation was complete at potentials positive to -10 mV and removed by hyperpolarization. 6. Inactivation of the transient current was slowed and accelerated in oxidizing and reducing conditions, respectively, confirming the involvement of an inactivating 'ball and chain' peptide. 7. R-PIA increased the transient current. The steady-state inactivation curve was shifted towards more positive potentials without changing the activation kinetics. Pretreatment with pertussis toxin (1 microgram ml-1) blocked the response to R-PIA. 8. It is concluded that frog melanotrophs possess an A-type current that is likely to play an important role in excitability. This current, which is directly modulated by A1 adenosine receptors through a Gi/G(o) protein, appears to be responsible for the inhibitory effects of adenosine on electrical activity.

4-Aminopyridine↗
PubMed1995

Activation of dopamine D4 receptor inhibits an L-type calcium current in cerebellar granule cells.

The functions of the D4 receptor, a newly cloned D2-like receptor, as well as the identity of cells expressing it, are still poorly defined. Using quantitative polymerase chain reaction we detected the messenger RNA of the D4, but not other D2-like receptor, in cultured granule cells from neonatal rat cerebellum. In these neurons, dopamine reduced high-voltage-activated calcium current, with a pharmacology corresponding to that of the D4 receptor. The response declined from one to three days, when calcium currents were mostly sensitive to nifedipine, to 15 days, when nifedipine-insensitive calcium currents were also present and D4 receptor messenger RNA had declined. The dopamine response was abolished after pretreatment of the cells by pertussis toxin, was potentiated and made irreversible by infusion of guanosine 5'-O-(3-thiotriphosphate) but persisted in the presence of cyclic AMP and isobutylmethylxanthine. These results indicate the presence in the neonatal cerebellum of a functional D4 receptor inhibiting an L-type calcium current, an action involving a Gi/Go protein but independent from adenylate cyclase inhibition.

Adenylyl Cyclases↗
PubMed1994

Inhibitory effect of adenosine on electrical activity of frog melanotrophs mediated through A1 purinergic receptors.

1. The effects of adenosine were studied in cultured frog melanotrophs by the patch-clamp technique. 2. In cell-attached experiments, most cells responded to adenosine (50 microM) by a reversible inhibition of action current discharges without any apparent desensitization. 3. In whole-cell experiments, adenosine provoked a hyperpolarization accompanied by a depression of spontaneous action potentials and a decrease in membrane resistance. When adenosine was repeatedly applied, tachyphylaxis was observed. Addition of GTP (100 microM) in the intracellular solution augmented the percentage of cells hyperpolarized by adenosine, and the duration and amplitude of the hyperpolarization, and prevented the tachyphylaxis. 4. Pretreatment with pertussis toxin (1 microgram ml-1) blocked adenosine-induced inhibition. 5. In cells dialysed with the non-hydrolysable GTP analogue GTP gamma S (100 microM), adenosine caused a sustained, strong hyperpolarization and an irreversible inhibition of spikes. 6. The effect of adenosine was mimicked by the A1 receptor agonist R-PIA (R-N6-phenylisopropyl-adenosine; 50 microM) and blocked by the A1 receptor antagonist CPDPX (8-cyclopentyl-1,3-dipropylxanthine, 50 microM). The A2 receptor antagonist CGS15943 (9-chloro-2-(2-furanyl)-5,6-dihydro-1,2,4-triazolo[1,5-c] quinazoline-5-imine; 50 microM) did not affect the adenosine-induced response. 7. The results suggest that, in frog melanotrophs, adenosine exerts a direct hyperpolarizing effect accompanied by blockage of spontaneous action potentials. The effect of adenosine is mediated through A1 receptors coupled to a Gi/o protein.

Action Potentials↗
PubMed1994

Melanostatin (NPY) inhibited electrical activity in frog melanotrophs through modulation of K+, Na+ and Ca2+ currents.

1. Melanostatin, a thirty-six amino acid peptide recently isolated from the frog brain due to its ability to inhibit alpha-melanocyte-stimulating hormone (alpha-MSH) release, is the amphibian counterpart of mammalian neuropeptide Y (NPY). The effect of synthetic melanostatin on the bioelectrical activity of cultured frog melanotrophs was studied in 124 cells by using the whole-cell patch-clamp technique. 2. In current-clamp experiments, melanostatin (1 microM) provoked a reversible hyperpolarization and a suppression of spontaneous action potentials. In some cells the hyperpolarizing response was absent, but an arrest of spike firing still occurred. 3. Melanostatin-induced hyperpolarization was associated with a decrease in membrane resistance. In voltage-clamp experiments, melanostatin induced an outward current at a constant command potential. This hyperpolarizing outward current appeared to be carried by potassium ions. 4. Cell dialysis with the non-hydrolysable GTP analogue guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S) sustained the outward current produced by melanostatin. Dopamine (1 microM), which generates a similar hyperpolarizing outward current in frog melanotrophs, was not capable of increasing the current provoked by melanostatin and sustained by GTP gamma S. 5. Melanostatin also modulated voltage-operated currents. The amplitude of voltage-activated potassium current was increased by 30%. 6. Melanostatin reduced the fast sodium current. This inhibitory effect was rather persistent compared to the other modulated currents. 7. Melanostatin markedly scaled down high voltage-activated N- and L-like calcium currents. The activation kinetics of these two calcium currents were not altered by the peptide. 8. Pretreatment of melanotrophs with pertussis toxin (1 microgram ml-1) blocked melanostatin-induced inhibition of N- and L-like calcium currents. 9. It is concluded that the NPY-related peptide melanostatin generates a very complex pattern of electrical responses in frog melanotrophs, including hyperpolarization and modulation of voltage-activated currents underlying action potentials. G proteins appear to mediate at least part of these effects.

Action Potentials↗
PubMed1994

Characterization of the GABA-induced current in frog pituitary melanotrophs.

The molecular mechanisms regulating GABAA receptor activity in cultured frog melanotrophs were studied using the patch-clamp technique. In the whole-cell configuration, application of GABA evoked a dose-related increase of inward chloride currents. The ED50 value, estimated from the sigmoidal dose-response curve was 2 x 10(-6) M and the Hill coefficient was 1.55. The amplitude of the GABA-induced current decayed with time. Kinetics analysis of the desensitization revealed that the time-course of the current decrement was fitted by one exponential. Graded doses of GABA or association of GABA with the benzodiazepine receptor agonist flunitrazepam accelerated the desensitization process. In contrast, the time-course of the current did not significantly vary at different holding potentials. In the outside-out configuration, GABA was found to activate channels which displayed three unitary conductance levels (8, 15 and 30 pS). The channel openings of the more frequent conductance level (30 pS) exhibited short and long lasting open states (1.2 and 28.3 ms at -60 mV). Altogether these data reveal that frog melanotrophs possess a single population of GABAA receptors which interconvert into a higher affinity state in the presence of benzodiazepine receptor agonists. Two GABA molecules must bind to the receptor to trigger long lasting channel openings. In addition, the activity of the GABAA receptor appears to be independent of the accumulation of intracellular chloride ions.

Animals↗
PubMed1994

Adrenaline induces hyperpolarization in frog pituitary melanotrophs through activation of potassium channels.

A patch-clamp study was conducted on cultured frog pituitary melanotrophs, in order to investigate the effects of adrenaline on the electrical activity of these cells. In the whole-cell configuration, adrenaline (1 microM) caused hyperpolarization that was accompanied by a fall in membrane input resistance and a blockage of spontaneous action potentials. Under voltage clamp, adrenaline elicited a net-outward current. The hyperpolarization became undetectable at a command voltage of -100 mV which corresponded to the equilibrium potential of potassium ions. The effect of adrenaline on membrane potential and spontaneous activity was blocked by the alpha 2-adrenergic receptor antagonist yohimbine (1-10 microM) but could not be mimicked by the alpha 2-adrenergic agonist clonidine (1-10 microM). In the cell-attached configuration, exposure of the extra-patch membrane to adrenaline increased the occurrence of single-channel currents with a slope conductance of 100 pS. The deduced reversal potential of these currents corresponded to the equilibrium potential of potassium ions. These results suggest that frog melanotrophs display an alpha 2-adrenergic receptor subtype coupled to potassium channels involved in hyperpolarization.

Adrenergic alpha-Antagonists↗