Changes In Membrane Conductance Research Articles

Changes in spontaneous otoacoustic emissions produced by acoustic stimulation of the contralateral ear

Spontaneous otoacoustic emissions (SOAEs) were measured in human ear canals before, during and after presentation of tonal stimuli to the contralateral ear. Stimuli were presented in 1 8 octave steps from 2 octaves below to 1 octave above the SOAE frequency at sound levels below the observed contralateral acoustic reflex threshold. For certain conditions there was an abrupt upward frequency shift at stimulus onset. For a fixed level the effect was frequency selective; the maximum frequency shift was obtained with tones approximately 1 2 octave below the SOAE. SOAE amplitude usually decreased but in some cases increased or remained unchanged. When amplitude changes were observed, the maximum shifts were observed for tones at or near the SOAE frequency. Changes in SOAEs were not observed for stimulus levels below 60 dB SPL. The effect is believed to be mediated by medial efferent neurons of the uncrossed olivocochlear bundle which arise in the medial region of the superior olivary complex and terminate on outer hair cells (OHCs). These results support those models which attribute SOAE generation to OHCs, and are indicative of an efferent influence on cochlear mechanics. A simple model is presented that proposes that efferent activity alters the tuning of the emission generator by causing changes in OHC membrane conductance.

Properties of membrane ion conductances evoked by hormonal stimulation of guinea-pig and rabbit isolated hepatocytes

Membrane conductance changes evoked in isolated guinea-pig or rabbit hepatocytes by hormonal stimulation were studied with the whole-cell patch clamp technique. In Cl-containing solutions, noradrenaline (NA), ATP or angiotensin II (AII) evoked an increase of conductance to both K (GK) and Cl (GCl) ions. Activation of GK occurred after a delay of several seconds and was sustained in the presence of hormone. Activation of GCl was transient, lasting several seconds, and arose either at the same time or shortly after the increase in GK. Conductances showed an initial rapid rise and slow oscillatory changes during maintained hormone application. The NA-induced current reversed at -19 mV in Cl solutions, between the equilibrium potentials for chloride (ECl = 0 mV) and potassium ions (EK = -85 mV), and at -75 mV, near EK, in Cl-free solution. In both conditions whole-cell current-voltage curves were linear in the range -100 mV to +40 mV. The conductance increase produced by NA to Cl- ions was about 50 nS, that to K+ ions was 6 nS. The potassium conductance increase was abolished by the polypeptide toxin apamin (50 nM). An increase in membrane current noise was associated with NA-evoked outward K+ current and blocked by apamin. Spectral analysis gave estimates of the elementary K channel conductance of 1.7 pS. Power spectra were fitted by two Lorentzian components, with average half-power frequencies of 2 and 190 Hz. These results are discussed in relation to the single-channel properties and indicate that the open probability of K channels during the NA response is high. In Cl solutions, with apamin to block the K conductance, no increase in current noise was detected during the large Cl conductance evoked by NA. This suggests either that Cl channels are of very low unitary conductance (less than 1 pS) or that Cl transport is due to a membrane carrier. The complex time-course of hormonally evoked conductances is not due to the properties of ion conductances per se but probably to underlying changes of intracellular second-messenger concentration.

Norepinephrine selectively reduces slow Ca2+- and Na+-mediated K+ currents in cat neocortical neurons.

1. The effects of norepinephrine (NE) and related agonists and antagonists were examined on large neurons from layer V of cat sensorimotor cortex ("Betz cells") were examined in a brain slice preparation using intracellular recording, constant current stimulation and single microelectrode voltage clamp. 2. Application of NE (0.1-100 microM) usually caused a small depolarization from resting potential; hyperpolarizations were rare. Application of NE reversibly reduced rheobase and both the Ca2+- and Na+-dependent portions of the slow afterhyperpolarization (sAHP) that followed sustained firing evoked by constant current injection. The faster Ca2+-dependent medium afterhyperpolarization (mAHP), the fast afterhyperpolarization (fAHP), the action potential, and input resistance were unaffected. 3. The changes in excitability produced by NE application were most apparent during prolonged stimulation. The cells exhibited steady repetitive firing to currents that were formerly ineffective. The slow phase of spike frequency adaptation was reduced selectively and less habituation occurred during repeated long-lasting stimuli. The relation between firing rate and injected current became steeper if firing rate was averaged over several hundred milliseconds. 4. During voltage clamp in TTX, NE application selectively reduced the slow component of Ca2+-mediated K+ current. The faster Ca2+-mediated K+ current was unaffected, as were two voltage-dependent, transient K+ currents, the anomalous rectifier and leakage conductance measured at resting potential. Depolarizing voltage steps in the presence of Cd2+ revealed an apparent time- and voltage-dependent increase of the persistent Na+ current after NE application. The voltage-clamp results suggested ionic mechanisms for all effects seen during constant current stimulation except the depolarization from resting potential. The latter was insensitive to Cd2+ and TTX and occurred without a detectable change in membrane conductance. 5. NE application did not alter Ca2+ spikes evoked in the presence of TTX and 10 mM TEA. Inward Ca2+ currents examined during voltage clamp in TTX (with K+ currents reduced) became slightly larger after NE application. We conclude that NEs reduction of the slow Ca2+-mediated K+ current is not caused by reduction of Ca2+ influx. 6. Effects on membrane potential, rheobase, and the sAHP were mimicked by the beta-adrenergic agonist isoproterenol, but not by the alpha-adrenergic agonists clonidine or phenylephrine at higher concentrations.(ABSTRACT TRUNCATED AT 400 WORDS)

Agonist‐induced changes in cell membrane capacitance and conductance in dialysed pancreatic acinar cells of rats.

1. Single acinar cells enzymatically isolated from the rat pancreas were subjected to tight-seal whole-cell recordings. Changes in cell membrane capacitance and conductance were simultaneously recorded using a phase-sensitive detection method. 2. Acetylcholine (ACh, 0.05-0.5 microM) and cholecystokinin octapeptide (CCK-8, 10-50 pM) concomitantly induced transient increases in cell membrane current, capacitance and conductance only when cytosolic Ca2+ was weakly chelated by EGTA (70 microM). These responses were prolonged when the cells were dialysed with a solution containing GTP gamma S (a stable analogue of GTP, 50-100 microM), whereas they were inhibited by dialysing with that containing GDP beta S (a stable analogue of GDP). These results suggest that a type of guanine-nucleotide-binding protein (G-protein) could be involved in ACh- or CCK-receptor signalling. 3. The ACh- or CCK-induced responses (with or without GTP gamma S in the cytosol) were all abolished when a high dose of EGTA (1-2 mM) was injected into the acinar cells. In addition, A23187, a calcium ionophore, induced sustained responses when the cytosolic Ca2+ was weakly buffered by 70 microM-EGTA. These results suggest that the secretagogues regulate the changes in cell membrane capacitance and conductance via an increase and decrease of cytosolic Ca2+ concentration. 4. Oscillatory changes in cell membrane conductance and capacitance were consistently observed even without applying secretagogues when the cells were dialysed with a solution containing GTP gamma S (50-100 microM) and cytosolic free Ca2+ ions weakly buffered at about 10(-6) M with a low dose of EGTA and CaCl2. 5. The peak amplitude of changes in cell membrane capacitance induced by ACh or CCK-8, with or without GTP gamma S in the cytosol, varied between 200 and 1000 fF, thereby suggesting that 20-100 zymogen granules can fuse with the luminal cell membrane in response to these agonists in rat exocrine pancreatic acinar cells.

Effect of leukotriene C4 on electromechanical activity and Ca2+ uptake in taenia coli.

The actions of leukotriene C4 (LTC4) on electromechanical activity and 45Ca2+ uptake in guinea pig taenia coli were investigated. The contractile action of LTC4 was abolished by the removal of extracellular Ca2+. LTC4 concentrations eliciting a maximal contraction in normal medium produced no response in preparations depolarized with KCl. In single sucrose gap studies, LTC4 increased both the frequency of electrical spiking and tension. These effects were blocked by the dihydropyridine Ca2+-channel antagonist PY 108-068 and by the leukotriene receptor antagonist FPL 55712. In double sucrose gap experiments, LTC4 caused a small depolarization without measurable change in membrane conductivity; increased spontaneous electrical activity was again accompanied by an increase in tension. LTC4 caused a detectable increase in 45Ca2+ uptake only at extracellular Ca2+ concentrations less than 1 mM, and this was again inhibited by PY 108-068 or FPL 55712. It is concluded that the contractile effects of LTC4 in guinea pig taenia coli occur as a consequence of its ability to open voltage-sensitive Ca2+ channels, an effect that may occur independently of membrane depolarization.

Voltage clamp studies of fertilization in sea urchin eggs: II. Current patterns in relation to sperm entry, nonentry, and activation

Following attachment of a sperm to the surface of a sea urchin egg clamped at a membrane potential ( V m) more positive than +17 mV, no changes in membrane conductance can be detected, the sperm does not enter the egg, and no morphological changes can be detected. At V m from +17 to −100 mV three characteristically different types of current profiles are observed: Type I are activation currents in eggs penetrated by a sperm. These have three phases, which occur in all eggs clamped at V m from +17 to −20 mV and in decreasing percentages at clamped V m more negative than −20 mV (to −75 mV). Complete fertilization envelopes are elevated, relatively large mound-shaped fertilization cones form, and the eggs develop to normal embryos. Type II are sperm transient currents in eggs not penetrated by a sperm, the eggs otherwise remaining in the unfertilized state. These transients are simpler and shorter than type I currents, and are observed only at clamped V m more negative than −20 mV. Type III are modified activation currents in eggs not penetrated by a sperm. These have three phases, are observed only at clamped V m more negative than −20 mV, and are the only type of activation current seen at clamped V m more negative than −75 mV. Complete fertilization envelopes are elevated, the fertilization cones are small and filament-like, and the eggs fail to cleave. We conclude that (a) the sperm transient currents (type II) and phase 1 of the activation currents (types I and III) are similar events generated by a sperm-initiated localized conductance increase, (b) the abrupt decrease of current which terminates the sperm transients and phase 1 of type III currents results from a turnoff of the sperm-induced conductance increase and signals that the sperm will not enter the egg, and (c) the occurrence of phase 2 during an electrophysiological response induced by a sperm indicates that the egg is activating.

Passive electrical properties and electrogenic sodium transport of cultured guinea-pig coronary endothelial cells.

1. Coronary endothelial cells were isolated from adult guinea-pig hearts (Nees, Gerbes & Gerlach, 1981) and the electrical properties of primary cultures were studied using the tight-seal whole-cell recording mode of the patch clamp technique. 2. On the third or fourth day in culture whole-cell clamp records from single coronary endothelial cells were obtained at 37 degrees C. The resting potential was -33 +/- 6 mV (n = 10). The membrane time constant determined with rectangular current pulses was 68 +/- 22 ms (n = 10). 3. In voltage clamp experiments, no time-dependent membrane conductance changes were found in the range -80 to +40 mV. The current-voltage relation was linear in normal physiological salt solution containing 5.4 mM-K+. The input resistance was 1.7 +/- 0.4 G omega. When the external K+ concentration was increased to 116 mM the cells depolarized to about -3 mV and the clamp currents showed marked inward rectification. 4. Between days four and seven in culture the endothelial cells formed confluent monolayers which showed the characteristic 'cobblestone' morphology. The input resistance of cells in a monolayer was 8 +/- 3 M omega, i.e. a factor of 200 lower than that found in single cells. It was concluded that the cells in the confluent monolayer are coupled electrically by gap junctions. 5. Exposure of coronary endothelial cells to K+-free solution for 5 min produced a depolarization of about 8 mV. Upon readmission of normal external K+ the cells transiently hyperpolarized by up to 20 mV. This transient hyperpolarization decayed with a time constant of 1.9 +/- 0.3 min. 6. The transient hyperpolarization could be abolished by application of 2 x 10(-4) M-dihydro-ouabain (DHO). Application of DHO in the steady state produced a depolarization of 8 +/- 1 mV. From these findings it was concluded that coronary endothelial cells possess an electrogenic sodium pump which contributes about -8 mV to the resting potential. 7. From the passive electrical properties of single cells and the morphological data available it was calculated that endothelium in situ may have a large electrical space constant, probably between 250 and 550 microns. 8. The functional implications of the large space constant of the endothelial monolayer are discussed. It is suggested that intra-endothelial conduction of electrical signals from capillaries to the resistance vessels may be involved in the local regulation of blood flow in the intact heart.

The influence of pH on membrane conductance and intercellular resistance in the rat lens.

1. The conductance of the rat lens was measured using a two-internal-microelectrode technique. The voltage response to a step of current consisted of two components arising from bulk and membrane resistance respectively. 2. The potassium permeability was calculated by applying Goldman theory to 86Rb+ efflux data. 3. The internal pH (pHi) and internal free calcium (pCai) were measured directly using single- and double-barrelled ion-sensitive microelectrodes. 4. Lens pHi was 6.9 in control solution (external pH, pHo = 7.3) and was reduced on lowering pHo. The presence of propionate or 100% CO2 in the external solution accentuated this effect. 5. Internal acidification was accompanied by a depolarization of membrane potential, an increase in membrane and cell-to-cell resistance and a decrease in potassium permeability. The acidification had no effect on pCai. 6. The intracellular pH was increased by perifusing with trimethylamine or NH4Cl. Both treatments induced a membrane depolarization with little change in potassium permeability. Subsequent removal of NH4Cl led to a sustained decrease in pHi. 7. In every case where pHi decreased, the changes in membrane potential and conductance could be explained largely on the basis of a decrease in potassium permeability. The concomitant increase in cell-to-cell resistance was less pronounced and probably insufficient to uncouple the lens system.

Immunoglobulin E mediated membrane conductance changes in rat basophilic leukemia cells.

Electrophysiological effects of anaphylactic stimulation of rat basophilic leukemia cells (RBL-2H3) were studied using conventional microelectrodes. Stimulation of passively sensitized cells by anti-immunoglobulin E resulted in hyperpolarization followed by depolarization. These changes in membrane polarization were associated with a decrease in input membrane resistance. No effect of anaphylactic stimulation was seen in Ca2+-free solution or when Ca2+ influx was blocked by Co2+, but it was mimicked by the Ca2+ ionophore A-23187. This suggests that the changes in ionic conductances were associated with calcium influx. These results support the hypothesis that membrane conductance changes are involved in the stimulus-secretion process of the RBL-2H3 cells.

The effect of temperature on neuromuscular transmission in the main caudal artery of the rat.

1. Excitatory junction potentials (EJPs) recorded in isolated segments of the proximal main ventral artery of the rat tail were reduced in amplitude and prolonged in time course as temperature was lowered from 35 to 15 degrees C. 2. The slow depolarization that followed the EJPs after supramaximal or repetitive perivascular stimulation was markedly slowed in time course, but little affected in amplitude, as temperature was lowered. 3. The time constant (tau EJP) of the exponential decay phase of the EJP recorded from cells deep in the media was similar to the membrane time constant, so that the increase in tau EJP at low temperatures is consistent with a decrease in membrane conductance. 4. The value of tau EJP was also prolonged if the EJP was evoked at the time of the peak of the slow depolarization; this effect was blocked by idazoxan (10(-7) M) but not by prazosin (10(-6) M). 5. During repeated short bursts of high-frequency stimulation, action potential initiation was facilitated by both the prolongation of EJPs and summation of slow depolarizations; these effects were greater at 25 than at 35 degrees C. 6. The interactions between EJPs and alpha-adrenoreceptor-mediated membrane conductance changes are considered with respect to the electrical events occurring during sympathetic neuromuscular transmission at the natural temperatures of the rat tail.

Modification of K+ conductance of heart cell membrane by BRL 34915.

1. The effect of the K+ channel agonist BRL 34915 on membrane conductance was investigated in isolated guinea-pig cardiac myocytes. 2. BRL 34915 reduced the duration of the transmembrane action potential and slightly increased the membrane resting potential in a concentration-dependent manner. 3. BRL 34915 removed the rectification in the steady-state current-voltage relationship. At membrane potentials more negative than the K+ equilibrium potential, membrane conductance was reduced. In the presence of 10(-4) mol/l BRL 34915, the current-voltage relationship was linear, i.e. of an ohmic type. 4. The BRL 34915-mediated change in membrane conductance was susceptible to the K+ channel blockers BaCl2 and tetrahydroaminoacridine. 5. In conclusion, BRL 34915 modifies K+ conductance in the cardiac cell membrane. The precise nature of the K+ conductance change remains to be elucidated.

Stimulation of NaCl secretion in the rectal gland of the dogfish Squalus acanthias

1. 1. The rectal gland of the dogfish ( Squalus acanthias) secretes NaCl when stimulated by a hormone related to vasointestinal peptide. 2. 2. Patch clamp and microelectrode techniques are used to examine the changes in membrane conductances occurring with hormonal stimulation. 3. 3. The conductance of the “resting” cell is dominated by basolateral K + channels. 4. 4. Hormonal stimulation “opens” apical Cl − channels. 5. 5. This opening of apical chloride channels appears to be mediated by cAMP-dependent phosphorylation of pre-existing closed channels.

Muscarinic M2 receptors on cardiac ganglion neurons of the guinea-pig heart.

Intracellular recordings were made from neurons of the cardiac ganglion of the guinea-pig heart. Application of muscarinic agonists produced a depolarization in the majority of neurons. The change in membrane conductance underlying the depolarization consisted of an initial low-conductance phase followed by a high-conductance phase. The first component was probably due to suppression of a potassium conductance, and the second was likely generated by activation of a chloride conductance. Both components of the depolarization were blocked by pirenzepine with an antagonist dissociation constants (KD) of 200-900 nM, suggesting that both are mediated through a muscarinic M2 receptor.

Presynaptic K-channel blockade counteracts the depressant effect of adenosine in olfactory cortex

Slices of isolated olfactory cortex from guinea-pig have been used to study the action of adenosine at synapses between axons of the lateral olfactory tract and neurons in the olfactory cortex. Adenosine depressed the excitatory postsynaptic potential, and, with paired or multiple stimuli, the reduced excitatory postsynaptic potentials in adenosine showed more synaptic facilitation. Very small excitatory postsynaptic potentials which were estimated not to be affected by postsynaptic membrane conductance changes were highly sensitive to adenosine. Both observations indicate a presynaptic action of adenosine. To test whether a conductance increase to potassium ions mediated adenosine action, the K-channel blockers, 3,4-diaminopyridine (1–100 μ mol/l) or 4-aminopyridine (100–500 μmol/l) were applied or Cs partially substituted for K. These substances reduced or prevented adenosine from having its depressant effect on synaptic transmission. These particular K-channel blockers also prolonged the action potential propagating along the lateral olfactory tract. When the increased excitability was counteracted by high Mg or low concentrations of tetrodotoxin, 3,4-diaminopyridine still blocked adenosine action. UO 2 ions prolonged the lateral olfactory tract action potential without blockade of K-conductance, but still supported an adenosine depression of the excitatory postsynaptic potential. Veratridine also supported the adenosine depression. These observations suggest that the action of 3,4-diaminopyridine on adenosine was not solely the result of increased tissue excitability. In contrast, tetraethylammonium (20 mmol/l), Ba (0.5–4 mmol/l) or Rb replacement for K had a negligible effect on the duration of the presynaptic action potential and had no effect on the depressant action of adenosine. These data are compatible with the idea that adenosine enhances an aminopyridine-sensitive potassium conductance in nerve terminals and changes in Ca influx are consequential to this.

Postsynaptic membrane shifts during frequency potentiation of the hippocampal EPSP.

1. In some classes of central neurons, repetitive synaptic stimulation induces substantial changes in the postsynaptic membrane, in conjunction with robust frequency potentiation of the excitatory postsynaptic potential (EPSP). However, the nature and time course of these postsynaptic membrane shifts, or their possible contributions to EPSP frequency potentiation (e.g., by altering driving force or current pathways), have not been examined extensively. We therefore studied the simultaneous patterns of change in composite EPSP amplitude, postsynaptic input resistance (Rin), and postsynaptic membrane potential during a 4-min train of 10-Hz monosynaptic stimulation in CA1 neurons of hippocampal slices. Slices were maintained in media containing either control (4 mM) or high (6.5 mM) concentrations of K+. 2. Potentiation of the EPSP, hyperpolarization of the membrane, and a decline of Rin, all developed rapidly during 10-Hz synaptic stimulation; these responses reached maximal levels by 5-15 s of the stimulation train. In most cells, a membrane depolarization phase occurred between 15 and 45 s of stimulation, followed by rehyperpolarization by 1 min of stimulation. During the depolarization phase, both EPSP potentiation and the decline in Rin remained near maximal. No significant differences were seen as a function of K+ concentrations. 3. These results show that hyperpolarization is not invariably associated temporally with EPSP frequency potentiation. Moreover, if driving force and membrane conductance changes are assumed to be approximately similar in large dendrites and soma, then the increase in driving force due to membrane hyperpolarization was not sufficient to account for the three- and fourfold increases in EPSP amplitude seen during frequency potentiation. Further, based on similar assumptions and on dendritic models of EPSP attenuation, the decline in Rin should reduce EPSP amplitude at the dendritic synaptic site and, to a proportionately greater extent, at the soma. 4. Studies in which the membrane was hyperpolarized with injected current to approximately the IPSP reversal potential, or in which bicuculline methiodide was applied to the slices, indicated that depression of the IPSP by repetitive stimulation did not account for frequency potentiation of EPSP amplitude. 5. These data are therefore consistent with the conclusion that the frequency potentiation of composite EPSPs in central neurons depends on presynaptic mechanisms, rather than on generalized postsynaptic changes. However, our findings do not rule out localized postsynaptic changes in receptors or spines as possible contributing factors.(ABSTRACT TRUNCATED AT 400 WORDS)

Membrane conductance oscillations in astrocytes induced by phorbol ester.

Glial cells in the central nervous systems (CNS) have complex functions which are difficult to decipher because of the intimate intertwining of glial cells with neurons. We have therefore developed an essentially neuron-free preparation of CNS astrocytes in the kainic acid lesioned hippocampal slice. With this preparation we have examined the effect of activating protein kinase C in astrocytes with a phorbol ester, TPA (12-O-tetradecanoyl-phorbol-13-acetate). In most cells, TPA induced rhythmic oscillations (0.1-3.0 Hz) of membrane potential which were typically 5-10 mV in amplitude and were associated with increases of up to eightfold in input resistance during the depolarizing phase. These large changes in membrane conductance are the first reported observations of endogenously generated conductance changes in astrocytes of the mammalian CNS and they could influence excitability of surrounding neurons, possibly by altering extracellular ion concentrations.

Dissociation of acetylcholine- and cyclic GMP-induced currents inXenopu oocytes

In Xenopus follicular oocytes, activation of muscarinic receptors evokes a slow potassium current (H-response); a similar current is evoked by intracellular injection of cyclic guanosine 3',5'-monophosphate, cGMP (Dascal et al. 1984). We have tested the hypothesis that cGMP may be the second messenger that mediates the opening of K channel by acetylcholine (ACh). ACh elevated the intracellular level of cGMP with a time course similar to that of the development of the muscarinic H-response; maximal increase in cGMP concentration above the control was about 0.2 pmole/oocyte. The amount of injected cGMP that produced a detectable K current ("threshold dose") varied between 0.5 and 3 pmole/oocyte. At low doses of cGMP, the slope of log dose-log response curve was about 2.5, suggesting involvement of a biochemical process with a positive cooperativity of at least 3. Higher doses of cGMP evoked, in addition to the outward current, an irregular, rapidly developing, long-lasting inward current, that never reached amplitudes comparable to those of ACh-evoked Cl currents. The K current elicited by cGMP was insensitive to elevation or depletion of external Ca. It was potentiated by isobutylmethylxanthine (IBMX). ACh strongly inhibited the cGMP-evoked K current when applied at the plateau of the latter. 4-Phorbol 12,13-dibutyrate (PDBu) (1 microM) rapidly and completely inhibited the cGMP response. It is concluded, that most of the results presented in this report contradict the hypothesis that cGMP is the intracellular mediator of ACh-induced changes in membrane conductance in the oocytes.

Push-pull effect of surround illumination on excitatory and inhibitory inputs to mudpuppy retinal ganglion cells.

1. Changes in membrane potential and conductance were measured in on-centre and off-centre ganglion cells during the responses to illumination of different portions of the receptive field. 2. In on-centre ganglion cells the sustained depolarizing response to steady illumination of the receptive field centre was associated with a net increase in conductance. In the presence of centre illumination, stimulation of the surround with an annulus of light caused a hyperpolarization and a net decrease in conductance, and the reversal potential of the light-evoked response was shifted in a negative direction. In the absence of centre illumination the same annular stimulus caused a hyperpolarization and a net increase in conductance. 3. In off-centre ganglion cells the sustained hyperpolarizing response to centre illumination was associated with a net increase in conductance. In the presence of centre illumination, stimulation of the surround with an annulus caused a depolarization and a net decrease in conductance, and the reversal potential of the light-evoked response was shifted in a positive direction. In the absence of centre illumination the same annulus caused a depolarization and a net increase in conductance. 4. The results indicate that illumination of the receptive field surround can affect both the excitatory and inhibitory sustained inputs to a given ganglion cell in a 'push-pull' manner, by decreasing the synaptic input that was increased by centre illumination and increasing the synaptic input of opposite sign. The relative effect of a given surround illumination on these two inputs, and hence the sign and magnitude of the net conductance change, varied with the amount of centre illumination.

Divalent cations directly affect the conductance of excised patches of rod photoreceptor membrane.

Phototransduction in rod cells is likely to involve an intracellular messenger system that links the absorption of light by rhodopsin to a change in membrane conductance. The direct effect of guanosine 3',5'-monophosphate (cGMP) on excised patches of rod outer segment membrane strongly supports a role for cGMP as an intracellular messenger in phototransduction. It is reported here that magnesium and calcium directly affect the conductance of excised patches of rod membrane in the absence of cGMP and that magnesium, applied to intact rod cells, blocks a component of the cellular light response. The divalent cation-suppressed conductance in excised patches showed outward rectification and cation-selective permeability resembling those of the light-suppressed conductance measured from the intact rod cell. The divalent cation-suppressed conductance was partly blocked by a concentration of the pharmacological agent L-cis-diltiazem that blocked all of the cGMP-activated conductance. Divalent cations may act, together with cGMP, as an intracellular messenger system that mediates the light response of the rod photoreceptor cell.

Actions of gamma-aminobutyric acid on rat supraoptic nucleus neurosecretory neurones in vitro.

1. Intracellular recordings were obtained from thirty-eight rat supraoptic nucleus (s.o.n.) neurosecretory neurones in perfused hypothalamic explants. Changes in membrane potential and conductance were monitored following application of gamma-aminobutyric acid (GABA), and related agonists and antagonists. 2. GABA depressed action potential discharge of all of thirty-five s.o.n. neurones tested and induced either membrane hyperpolarization or depolarization. Neurones that displayed membrane hyperpolarization in response to lower GABA concentrations (30-300 microM) demonstrated a biphasic membrane voltage change with a later depolarizing phase as a response to higher concentrations (up to 3000 microM). 3. GABA (10-3000 microM) induced a prominent concentration-dependent increase in membrane conductance in all neurones. The critical slope for the log-log plot of [GABA] vs. GABA-induced membrane conductance was 1.7, indicating co-operativity in the GABA receptor-induced conductance change. 4. Muscimol (0.3-30 microM) potently mimicked all the effects of GABA. Bicuculline (1-100 microM) antagonized the effects of GABA and muscimol in a competitive manner. 5. Glycine and taurine (1-10 mM) had weak effects, although comparatively similar to those of GABA. These actions were blocked both by bicuculline (100 microM) and by strychnine (1 microM). At higher concentrations (greater than 10 microM), strychnine also antagonized the actions of GABA. 6. In recordings with potassium-acetate-filled micropipettes, the reversal potential of hyperpolarizing membrane voltage responses to GABA was -72.5 +/- 1.5 mV in close agreement (+/- 5 mV) with the reversal potential of inhibitory post-synaptic potentials (i.p.s.p.s) recorded in the same neurones. Depolarizing responses to GABA reversed polarity at -50 +/- 1.6 mV. In recordings with KCl-filled micropipettes, voltage responses to GABA were always depolarizing and reversed near -40.0 +/- 4.3 mV. Similarly, reduction of the concentration of chloride ions in the perfusion medium from 134 to 10.4 mM induced a positive shift of the GABA reversal potential by 40-50 mV. 7. From measurements of input resistance (Rin) and cell time constant (tau O), input capacitance (Cin; representing total membrane capacitance) was calculated as 78.9 +/- 2.1 pF. During responses to GABA or muscimol, decreased Rin was accompanied by a linearly related decrease in tau o indicating that these substances had no effect on the membrane capacitance of s.o.n. neurones.(ABSTRACT TRUNCATED AT 400 WORDS)