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P A Fuchs

Publications and source records attributed to P A Fuchs.

35 records · Page 2Linked to original sources
PubMed1992

A novel cholinergic receptor mediates inhibition of chick cochlear hair cells.

The central nervous system provides feedback regulation at several points within the peripheral auditory apparatus. One component of that feedback is inhibition of cochlear hair cells by release of acetylcholine (ACh) from efferent brainstem neurons. The mechanism of hair cell inhibition, and the character of the presumed cholinergic receptor, however, have eluded understanding. Both nicotinic and muscarinic, as well as some non-cholinergic ligands can affect the efferent action. We have made whole-cell, tight-seal recordings from short (outer) hair cells isolated from the chick's cochlea. These are the principal targets of cochlear efferents in birds. ACh hyperpolarizes short hair cells by opening a cation channel through which Ca2+ enters the cell and subsequently activates Ca(2+)-dependent K+ current (Fuchs & Murrow 1991, 1992). Both curare and atropine are effective-antagonists of cholinergic inhibition at 3 microM, whereas trimethaphan camsylate and strychnine block at 1 microM. The normally irreversible nicotinic antagonist, alpha-bungarotoxin, reversibly blocked the hair cell response, as did kappa-bungarotoxin. The half-blocking concentration for alpha-bungarotoxin was 26 nM. It is proposed that the hair cell AChR is a ligand-gated cation channel related to the nicotinic receptor of nerve and muscle.

Acetylcholine↗
PubMed1992

Development of frequency tuning in the auditory periphery.

The peripheral auditory organ, the cochlea, acts as a spectral analyzer resolving the frequency components of sound. During development the cochlea first responds to loud low-frequency sounds, and only gradually acquires the adult pattern of increased sensitivity and an expanded high-frequency range. This evolution of function may result in part from the gradual maturation of hair cell properties.

Animals↗
PubMed1992

Cholinergic inhibition of short (outer) hair cells of the chick's cochlea.

Cochlear hair cells are thought to be inhibited by the release of ACh from efferent neurons. Several studies have implicated Ca2+ as a postsynaptic intermediary in hair cell inhibition, but its role remains unproven. We have made whole-cell, tight-seal recordings from single short hair cells (the avian analog of outer hair cells in the mammalian cochlea), isolated from the chick's cochlea, to determine the mechanism of cholinergic inhibition. These cells hyperpolarized upon exposure to ACh, although a brief depolarization preceded the much larger, longer-lasting hyperpolarization. In voltage clamp ACh evoked an outward current that reversed in sign near the K+ equilibrium potential. A small, transient inward current preceded the predominant outward current. The ACh-evoked K+ current depended on Ca2+ in the external saline, or could be prevented when the cell was dialyzed with the rapid Ca2+ buffer BAPTA. In BAPTA-loaded cells a residual inward current was seen. This activated with very little delay upon exposure of the cell to ACh and reversed near 0 mV membrane potential. Thus, the hair cell ACh receptor appears to be a nonspecific cation channel through which Ca2+ enters and triggers the opening of nearby Ca(2+)-activated K+ channels. However, the ACh-evoked K+ channels are not the same as the "maxi" K+ channels activated by Ca2+ influx through voltage-gated Ca2+ channels in these same cells.

Acetylcholine↗
PubMed1990

Preferential expression of transient potassium current (IA) by 'short' hair cells of the chick's cochlea.

We have made a comparative study of the membrane properties of tall and short hair cells isolated from a selected region of the chick's cochlea. Tall hair cells are analogous to inner cochlear hair cells of mammals, and like those, are presynaptic to the majority of afferent neurons in the cochlea. Short hair cells, like mammalian outer hair cells, are the postsynaptic targets of efferent neurons that inhibit the cochlea. Voltage-clamp recordings have revealed that short hair cells have an inactivating potassium (K) current, IA, whereas tall hair cells have little or none. Short hair cells are also sensitive to the cholinergic agonist carbachol, whereas tall hair cells are not. This pattern is in accord with the selective distribution of efferent cholinergic synapses in the cochlea. Although IA is completely inactivated at the resting potential of the short hair cells, cholinergic agonists can hyperpolarize these cells by as much as 30 mV. This hyperpolarization removes inactivation and allows IA to modulate subsequent voltage-dependent processes in short hair cells. It is concluded that IA could increase the high frequency response of the hair cell by decreasing membrane resistance and thus the membrane time constant after inhibition. This will be of particular importance to cochlear function if short hair cells produce voltage-dependent movements, as do mammalian outer hair cells.

Animals↗
PubMed1990

The acquisition during development of Ca-activated potassium currents by cochlear hair cells of the chick.

Voltage-clamp recordings were done on hair cells from a region of the chick's cochlea. In the adult, these cells have voltage-sensitive Ca currents and rapid, Ca-activated K currents that together support an electrical resonance, showing voltage oscillations at frequencies greater than 100 Hz. In embryos 14-days old (at one week before hatching) the same cells had a voltage-sensitive Ca current like that in adults, but a more slowly acting K current (of the delayed-rectifier type). In current-clamp they could generate only slowly repetitive action potentials. By two days before hatching, Ca-activated K currents were present. We suggest that the acquisition of Ca-activated K currents contributes to functional maturation of the chick's cochlea.

Action Potentials↗
PubMed1990

Potassium currents in hair cells isolated from the cochlea of the chick.

1. Potassium currents were characterized in tall hair cells of the chick's cochlea. Outward potassium currents were found to flow through two distinct classes of channels. 2. Individual hair cells were isolated from 200 microns long segments of the apical half of the chick's cochlea. Whole-cell voltage-clamp and current-clamp recordings were made from these cells. 3. Voltage responses to injected current ranged from high-frequency (100-250 Hz) oscillations in some cells, to slowly repetitive Ca2+ action potentials or slow oscillations (5-20 Hz) in others. 4. Ionic currents recorded in voltage clamp also varied in different hair cells. Cells with high-frequency voltage oscillations had rapidly activating Ca2(+)-dependent outward K+ current, IK(Ca). Cells that generated action potentials had slow delayed rectifier outward K+ current, IK, and inward rectifier current, IIR. All hair cells had inward Ca2+ current. 5. IK(Ca) activated positive to -45 mV. Tail currents reversed at the K+ equilibrium potential. This current was eliminated in Ca2(+)-free solutions, or when exposed to 10 mM-TEA. This outward current was fully activated within 1-3 ms at 0 mV. The whole-cell current was noisy and ensemble variance analysis suggested a single-channel conductance of 63 pS near 0 mV. 6. IK activated positive to -50 mV. Tail currents reversed at the K+ equilibrium potential. This current was not eliminated in Ca2(+)-free solutions, and was relatively resistant to external TEA. IK activated slowly, reaching peak values in 10-20 ms at 0 mV. This current showed little variance and the average single-channel conductance based on macroscopic noise near 0 mV was 8 pS. 7. External tetraethylammonium (TEA) or Ca2(+)-free saline eliminated the high-frequency voltage oscillations seen in many basal cells. In contrast TEA had little effect on slow action potentials (or low-frequency oscillations) seen in cells with IK. 8. IK(Ca) was prominent in hair cells originating 1.0-2.0 mm from the cochlear apex. IK and IIR dominated the membrane conductance of tall hair cells originating within 0.5 mm of the cochlear apex. 9. The frequency of voltage oscillation in apical cells was temperature-dependent, nearly doubling for each 10 degrees C rise in temperature. 10. IIR activated at membrane potentials negative to -75 mV. The average time constant of activation at -100 mV was 2 ms. Tail currents reversed at the K+ equilibrium potential and did not depend on the external Na+ concentration. IIR was blocked by 5 mM-Cs+ or 100 microM-Ba2+ in the external saline.

Action Potentials↗
PubMed1990

Calcium currents in hair cells isolated from the cochlea of the chick.

1. Calcium currents were characterized in tall hair cells isolated from the chick's cochlea to determine what types of Ca2+ channels existed and if these varied in cells with differing voltage responses to current injection. 2. Whole-cell, tight-seal recordings showed that the current-voltage relation of cochlear hair cells of the chick was dominated by K+ current. However, when outward K+ current was blocked it was found that all hair cells had a smaller, maintained inward current. 3. This inward current was a Ca2+ current since it required Ca2+ in the external medium, could also be carried by Ba2+, and was blocked reversibly by 5 mM-Co2+ and by Ni2+ and Cd2+ at micromolar concentrations. The Ca2+ channels were opened at membrane potentials positive to -50 mV, and the current was maximal near 0 mV. 4. The dihydropyridine BayK8644 (0.5 microM) produced a voltage-dependent increase of inward current. Ten micromolar nifedipine partially blocked the inward current. The outward Ca2(+)-activated K+ current was also reduced in the presence of 10 microM-nifedipine. These effects of dihydropyridines were completely reversible. 5. The Ca2+ current had rapid activation kinetics, reaching steady-state levels within 1 ms. If all outward currents were completely blocked the Ca2+ current showed no inactivation during depolarization lasting 200 ms. 6. No differences in voltage activation range, pharmacology, or kinetics of the Ca2+ current were found in tall hair cells from apical and basal regions of the cochlea. This is in contrast to the marked differences in K+ currents amongst cells from these two widely separated regions of the cochlea.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1988

Voltage oscillations and ionic conductances in hair cells isolated from the alligator cochlea.

Tall hair cells were isolated by enzymatic and mechanical dissociation from selected regions of the apical half of the alligator (A. mississippiensis) cochlea. Single cells were subjected to voltage-clamp and current-clamp using the tight-seal whole-cell recording technique. Most hair cells isolated from the apex of the cochlea produced slowly regenerative depolarizations or Na action potentials during current injection, whereas hair cells isolated from more basal regions usually produced voltage oscillations (ringing) in response to depolarizing current injection, an indication of electrical resonance. Resonant frequencies ranged from 50 to 157 Hz in different cells. The higher-frequency cells tended to have larger and more rapidly activating outward currents than did the lower-frequency cells. An inward Ca current and an outward Ca-activated K current were present in all hair cells. In addition, an inwardly rectifying current and a small, transient outward current were often seen. Thus, we conclude that an electrical tuning mechanism is present in alligator hair cells. The role of the ionic conductances in shaping hair cell responses to current injection, and the possible contributions of these electrical responses to cochlear function are discussed.

Action Potentials↗
PubMed1988

Electrical tuning in hair cells isolated from the chick cochlea.

Tall (inner) hair cells were isolated from specific locations in the chick cochlea. The electrical membrane properties of these cells were recorded using the tight-seal whole-cell technique. Depolarizing current steps elicited damped voltage oscillations that ranged in frequency from 100 to 250 Hz among cells from the middle third of the cochlea (basal cells). The current-voltage relation obtained under voltage clamp was dominated by calcium-activated potassium current in the voltage range over which these oscillations occurred. Tall hair cells isolated from the apical tip of the cochlea (apical cells) exhibited action potentials and lower frequency voltage oscillations (5-14 Hz) during depolarizing current steps. Outward currents in these cells were 20-fold slower than those found in the basal cells. These results suggest that electrical tuning of hair cells may play a role in determining the frequency selectivity of the chick cochlea.

Animals↗
PubMed1987

Tetrodotoxin-sensitive, voltage-dependent sodium currents in hair cells from the alligator cochlea.

We have used whole-cell patch clamp techniques to record from tall hair cells isolated from the apical half of the alligator cochlea. Some of these cells gave action potentials in response to depolarizing current injections. When the same cells were voltage clamped, large transient inward currents followed by smaller outward currents were seen in response to depolarizing steps. We studied the transient inward current after the outward current had been blocked by external tetraethylammonium (20 mM) or by replacing internal potassium with cesium. It was found to be a sodium current because it was abolished by either replacing external sodium with choline or by external application of tetrodotoxin (100 nM). The sodium current showed voltage-dependent activation and inactivation. Most of the spiking hair cells came from the apex of the cochlea, where they would be subject to low-frequency mechanical stimulation in vivo.

Alligators and Crocodiles↗
PubMed1985

Efferent modulation of hair cell tuning in the cochlea of the turtle.

Intracellular recordings were made from turtle cochlear hair cells in order to study the changes in their tuning properties resulting from electrical stimulation of the efferent axons. Efferent stimulation caused a reduction in the amplitude of the receptor potential at the hair cell's most sensitive or characteristic frequency, an increased amplitude at frequencies more than an octave below the characteristic frequency, and no change at very high frequencies. These differential effects resulted in a broadening of each cell's tuning curve, which, during maximal efferent stimulation degenerated from a sharply tuned resonance to a critically damped low-pass filter. Efferent alterations in tuning were also inferred from the oscillations in membrane potential produced by acoustic clicks or extrinsic currents. The quality factor (Q) of tuning, derived from the decay of the oscillations, was progressively reduced with synaptic hyperpolarizations up to about 5 mV in amplitude. A consequence of efferent action was that the wave forms of transient pressure changes were more faithfully encoded as changes in hair cell membrane potential. Hyperpolarization of a hair cell by steady current injection resulted in a lowering of its characteristic frequency and quality factor, and an increase in steady-state resistance. By comparison, for a given reduction in quality factor, efferent stimulation was associated with a smaller change in characteristic frequency. This difference is expected if the resonance is also damped by the shunting action of the synaptic conductance. Perfusion with perilymphs containing 0.5-15 mM of the potassium channel blocker, tetraethylammonium bromide (TEA) reduced the hair cell's frequency selectivity, whether assayed acoustically or with extrinsic currents. Lower TEA concentrations abolished the efferent inhibitory post-synaptic potential with only a minor change in tuning. TEA produced other effects different from efferent stimulation including (i) a lowering of the characteristic frequency, and (ii) a highly asymmetric receptor potential. These observations suggest that the efferents do not simply block membrane conductances associated with tuning. We conclude that the efferent modification of the shape of the tuning curve may be a composite result of the synaptic conductance and the hyperpolarization of the hair cell membrane.

Action Potentials↗
PubMed1984

Synaptic hyperpolarization and inhibition of turtle cochlear hair cells.

Intracellular recordings were made from turtle cochlear hair cells in order to examine the properties of the post-synaptic potentials evoked by electrical stimulation of the efferent axons. Single shocks to the efferents generated a hair cell membrane hyperpolarization with an average amplitude generally less than 1 mV and lasting for about 100 ms. With short trains of shocks, the size of the post-synaptic potential grew markedly to a maximum of 20-30 mV. The interaction between pairs of shocks separated by a varying interval was studied. For an interval of 4 ms, the response to the second shock was increased on average by a factor of 3 and the conditioning effect of the first shock decayed with a time constant of about 100 ms. We suggest the augmentation in response to trains of shocks may be partly due to facilitation of efferent transmitter release. The efferent post-synaptic potentials could be reversibly abolished by perfusion with perilymphs containing 3 microM-curare or atropine, and infusion of acetylcholine gave a transient membrane hyperpolarization. These observations are consistent with efferent action being mediated via a cholinergic synapse onto the hair cells. The post-synaptic potentials could be reversed in polarity by injection of hyperpolarizing currents through the recording electrode. The reversal potential was estimated as about -80 mV, 30 mV negative to the resting potential. Near reversal, a small brief depolarization was evident and may constitute a minor component of the synaptic response. The value of the reversal potential was unaffected by substitution of the perilymphatic chloride, but was altered in a predictable manner by changes in extracellular potassium concentration indicating that the post-synaptic potentials arise mainly by an increase in the permeability of the hair cell membrane to potassium ions. Throughout the post-synaptic hyperpolarization there was a reduction in the sensitivity of the hair cell to tones at its characteristic frequency. The desensitization, maximal for low sound pressures, varied in different cells from a factor of 1.6 to 28. At the peak of the largest synaptic potentials, the receptor potential remained negative to the resting potential with all but the loudest characteristic frequency tone s. We suggest that there are two factors in efferent inhibition; one a r duction in the receptor potential at the hair cell's characteristic frequency and the other a hyperpolarization of its membrane potential which should reduce the release of excitatory transmitter onto the afferent terminals.

Acetylcholine↗
PubMed1982

Efferent regulation of hair cells in the turtle cochlea.

Intracellular recordings were made from hair cells in the isolated cochlea of the turtle to characterize the inhibition achieved by the cochlea's efferent innervation. A short train of shocks delivered to the efferent axons produced in the hair cells slow hyperpolarizing synaptic potentials which could be reversed by shifting the membrane potential more negative than about -80 mV. Throughout the efferent hyperpolarization, there was a reduction of up to 25-fold in the amplitude of the receptor potential for tones presented at the hair cell's characteristic frequency. Efferent stimulation also was shown to degrade the cell's tuning properties. It is argued that the combined effects of the hyperpolarization and the loss in hair cell sensitivity could account for a threshold elevation of at least 70 dB in the auditory nerve fibres.

Action Potentials↗
PubMed1982

Chemical transmission between individual Retzius and sensory neurones of the leech in culture.

1. Chemical synaptic transmission develops between individual identified neurones dissected from leech ganglia and maintained in culture. Impulses in Retzius cells give rise to hyperpolarizing synaptic potentials in pressure (P) sensory cells. In suitable medium the potentials develop by 3 days and can be observed for more than 3 weeks. 2. The synaptic potentials occur after a synaptic delay, exhibit facilitation and depression and are reversed by hyperpolarization. The blocking effects of reduced calcium and raised magnesium concentrations in the bathing fluid provide additional evidence for the chemical nature of transmission. 3. An increase in chloride conductance is involved in the generation of the synaptic potential in the P cell. With high intracellular Cl in the post-synaptic cell, the synaptic potentials become reversed and amplified. The amplitudes of these reversed responses range from 1 to 20 mV with a falling phase lasting for seconds. 4. Changes in the membrane potential of the presynaptic cell that modify the amplitude and duration of the action potential influence the efficacy of transmission. In addition, impulses in Retzius cells initiated from hyperpolarized values of membrane potential evoke smaller synaptic potentials in the P cells than impulses arising from a depolarized level. 5. With neurones placed directly next to one another in the dish, maintained depolarization of the presynaptic Retzius cell in the absence of conducted action potentials gives rise to slow synaptic potentials in the P cells. In some pairs, the response in the P cell consists of a marked increase in 'noise'. 6. Injection of horseradish peroxidase into the Retzius cell reveals neurites with distinctive varicosities growing over the P cell.

Animals↗
PubMed1981

Membrane properties and selective connexions of identified leech neurones in culture.

1. Individual, identified neurones, dissected from the central nervous system of the leech and maintained in culture for several weeks, sprouted processes and formed synaptic connexions.2. The action potentials of isolated touch (T), pressure (P), nociceptive (N) cells and Retzius cells resembled those of their counterparts in situ, enabling them to be recognized unambiguously. Their input resistances were approximately 4 times greater than those of corresponding cells within the animal. In T, P and N cells trains of impulses were followed by a pronounced after-hyperpolarization, as in the animal.3. In certain cells, notably the L motoneurones, membrane properties became altered in culture. The current-voltage relation showed novel rectification and action potentials became much larger.4. Numerous neurites often extended for hundreds of micrometres from isolated neurones and ended in typical growth cones. Electron micrographs revealed that many fine axons were braided together to form thicker fascicles. Frequently, the processes were orientated between two neighbouring cells rather than at random. The fine structure of the cytoplasm, nucleus and organelles in cultured cells resembled those of their counterparts in situ. The glial cell that normally surrounds the neurones was, however, absent.5. Pairs of Retzius cells in culture usually became coupled electrically after about 6 days. Similarly L motoneurones became coupled in vitro. These junctions allowed current to pass in both directions and resembled those seen in the animal.6. Selective connexions were made by certain types of cells. Thus, P sensory neurones did not become coupled with Retzius cells but did develop electrical connexions with L motoneurones, as in the animal.7. Novel synaptic interactions not obvious in the animal could appear in culture. Retzius and L cells became electrically coupled and, in some instances where electrical coupling between Retzius cells failed to develop, chemically mediated inhibitory potentials became apparent.8. Isolated, identified leech neurones not only survive but regenerate processes and are capable of forming selective connexions in culture. The ability to define interactions between isolated pairs of cells offers the opportunity to explore in detail problems relating to synapse formation and cell-cell recognition.

Action Potentials↗
PubMed1980

Ionic basis of presynaptic inhibitory potentials at crayfish claw opener.

1. Intracellular recordings from the claw opener excitor axon of the crayfish, Procambarus clarkii, were obtained near the terminal arborizations of the axon on the surface of the opener muscle. Rest potential in the excitor axon averaged --80 mV over 20 cells. Action-potential amplitude and duration averaged 100 mV and 2 ms, respectively. 2. A single action potential in the opener inhibitor axon produces a hyperpolarizing synaptic potential (average amplitude 0.3 mV) in the excitor axon. The apparent reversal potential of this inhibitory synaptic potential is approximately 5 mV more negative than rest in control saline. No excitor axons were observed to have depolarizing synaptic potentials at rest. 3. A decrease in external chloride concentration from 240 to 24 mM causes the apparent reversal potential to depolarize an average of 12 mV, with no change in rest potential. In low-chloride saline, the synaptic potential evoked by stimulation of the inhibitor axon becomes depolarizing. 4. An increase in external potassium concentration from 5 to 10 mM causes the apparent reversal potential to depolarize by 16 mV; however, rest potential depolarizes by 10 mV. Low external potassium has the opposite effects, causing both rest potential and the apparent reversal potential to hyperpolarize. 5. Presynaptic inhibition at the Procambarus claw opener neuromuscular junction appears to be mediated by a hyperpolarizing synaptic potential. The results of these experiments suggest that chloride serves as the charge for the presynaptic potential. The evidence for a direct involvement of potassium as a charge carrier is equivocal due to Donnan equilibrium effects involving Cl.

Animals↗