A Biophysical Model of the Inner Hair Cell: The Contribution of Potassium Currents to Peripheral Auditory Compression

The term peripheral auditory compression refers to the fact that the whole range of audible sound pressure levels is mapped into a narrower range of auditory nerve responses. Peripheral compression is the by-product of independent compressive processes occurring at the level of the basilar membrane, the inner hair cell (IHC), and the auditory nerve synapse. Here, an electrical-circuit equivalent of an IHC is used to look into the compression contributed by the IHC. The model includes a mechanically driven transducer potassium (K⁺) conductance and two time- and voltage-dependent basolateral K⁺ conductances: one with fast and one with slow kinetics. Special attention is paid to faithfully implement the activation kinetics of these basolateral conductances. Optimum model parameters are provided to account for previously reported in vitro observations that demonstrate the compression associated with the gating of the transducer and of the basolateral channels. Without having to readjust its parameters, the model also accounts for the in vivo nonlinear IHC transfer characteristics. Model simulations are then used to investigate the relative contribution of the transducer and basolateral K⁺ currents to the nonlinear IHC input/output functions in vivo. The simulations suggest that the voltage-dependent activation of the basolateral currents compresses the DC potential for stereocilia displacements above approximately 5 nm. The degree of compression exceeds 2-to-1 and is similar for all stimulation frequencies. The AC potential is compressed in a similar way, but only for frequencies below 800 Hz. The simulations further suggest that the nonlinear gating of the transducer current is responsible for the expansive growth of the DC potential with increasing sound level (slope of 2 dB/dB) at low sound pressure levels.Both authors contributed equally to this work.     

Firing properties and dendrotoxin-sensitive sustained potassium current in vestibular nuclei neurons of the hatchling chick

To understand the emergence of excitability in vestibular nuclei neurons, we performed patch-clamp recordings on brain slices to characterize the firing pattern on depolarization and the underlying currents in principal cells of the chick tangential nucleus. This study, on 0- to 3-day-old hatchlings, distinguishes electrophysiologically one main group of principal cells based on their response to depolarizing current pulses (300-400 ms) in current-clamp recordings. This group (90%; n=29) displayed nonaccommodating, repetitive firing on depolarization. The remaining cells fired one action potential at the beginning of the current pulse and then accommodated. In voltage-clamp recordings, a low-threshold, sustained, dendrotoxin-sensitive (DTX; 200 nM) potassium current, I(DS), was identified in both cell groups. In the repetitively firing principal cells, the mean proportion of the DTX-sensitive sustained current contributing to the total outward current was less than 20%. This percentage is significantly less than that reported (45%) in a previous study performed in late chick embryos (E16), in which most of the cells (83%; n=89) were accommodating neurons. Tonic firing is an important electrophysiological feature characterizing most mature, second-order vestibular neurons, since it allows the neurons to process signals from behaviorally relevant inputs. Accordingly, this study contributes toward defining the emergence of the mature pattern of neuronal excitability and the ionic currents involved.     

The Changes of Potassium Currents in Rabbit Ventricle with Healed Myocardial Infarction

Epidemiologicalstudiesshowedthatpatientswithhealmyocardialinfarction (HMI)arethemainpopula tionofcardiacsuddendeath .CASTtrial[1 ] andSWORDtrial[2 ] hadaprofoundimpactonthepracticeofcardiolo gy,andthemortalitywassignificantlyhigherinpatientswithHMItreated…     

An improved loose patch voltage clamp method using concentric pipettes

A method for noninvasive voltage-clamp recording from large cells is described. A firepolished pipette having two concentric barrels is pushed against the cell membrane, thereby electrically isolating a circular patch subdivided into an inner and an annular outer region. Both regions are held isopotential, but current is collected from the inner region only. The method electrically simulates a high resistance seal between pipette and cell membrane, allowing accurate and rapid voltage-clamp recording under conditions where the seal resistances actually obtained are low (near 1 M). This is useful in applications where one wishes to avoid enzymatic treatment.We provide details of electrode construction and voltage-clamp electronics, and present results obtained from frog skeletal muscle and leech neurons. For sodium channels of frog muscle, extensive data were previously obtained with other methods. There is good agreement between the earlier results and the measurements presented here.     

Electrophysiological properties of cultured hippocampal neurons from Wistar Audiogenic Rats

The main goal of this work was to analyze the electrophysiological properties of cultured hippocampal neurons from a particular epileptic rat strain, called Wistar Audiogenic Rats (WAR). The whole-cell patch-clamp technique was used to record both active and passive membrane responses in an attempt to detect alterations in their characteristics in relation to controls from Wistar rats. Neurons from WARs show a significant reduction in the magnitude of the inhibitory GABAergic currents ( approximately 45%), in spite of maintaining a normal level of the excitatory glutamatergic currents. In addition, the magnitude of potassium currents, measured at +80 mV, is reduced by about 30% in comparison to controls. Surprisingly, we also found important changes in the passive cellular properties in WAR neurons such as membrane potential (-50.0 mV in WARs and -63.1 mV in controls) and input resistance (647 MOmega in WARs and 408 MOmega in controls). The changes described here, could be the basis of the neurophysiological and behavioral alterations present in these hyperexcitable animals, contributing to a better understanding of epileptogenesis in this particular animal model.     

Effects of binocular form deprivation on the excitatory post-synaptic currents mediated by N-methyl-D-aspartate receptors in rat visual cortex

PURPOSE: To investigate the effects of binocular form deprivation (BFD) on the excitatory post-synaptic currents (EPSCs) mediated by the N-methyl-D-aspartate (NMDA) receptor (NMDA-EPSCs), and the proportion of NMDA-EPSCs relative to glutamate receptor currents (glutamate-EPSCs) in rat visual cortex. METHODS: Binocular form deprivation was achieved by suturing the eyelids of Wistar rats at postnatal day (PD) 14, before eye-opening. Visual cortical slices (300 micro m) were prepared from normal and BFD Wistar rats aged PD 14, 21 and 28. Recordings were obtained in slices from layer II to IV using the whole-cell patch-clamp technique. Glutamate-EPSCs were isolated in the presence of bicuculline methiodide (20 micro mol/L) in the bathing medium, and NMDA-EPSCs were isolated with a combination of bicuculline methiodide (20 micro mol/L) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 20 micro mol/L). In addition, D,L-2-amino-5-phosphonovalerate (AP-5, 20 micro mol/L) was applied to study the NMDA-only mediated currents. For each cell, the ratio of peak NMDA to glutamate EPSCs was calculated. RESULTS: During visual development, the decay time constant of NMDA-EPSCs became shorter after eye-opening in normal rats (F = 5.949, P <0.05; PD 28 vs PD 14, P = 0.027), but not in rats with BFD (P > 0.05). The weighted time constant of NMDA-EPSCs in the visual cortex became shorter after the rats' eyes were opened in the normal group (F(2,37) = 4.727, P = 0.015; PD 28 vs PD 14, P = 0.035), but not in the BFD group (P > 0.05). However, the rise time constant and peak value of NMDA-EPSCs showed no significant changes in normal and BFD groups (P > 0.05). The ratio of NMDA-EPSCs to glutamate-EPSCs became gradually smaller with age in the normal rats (F = 4.661, P < 0.05; PD 28 vs PD 14, P = 0.025), but not in the BFD group (P > 0.05). CONCLUSIONS: These studies reveal that the proportion of NMDA-EPSCs relative to glutamate-EPSCs and the decay time constant of NMDA-EPSCs are influenced by BFD. These changes may reflect important experience-dependent modifications of neuronal synapses in visual cortex.     

Multiple signal pathways coupling VIP and PACAP receptors to calcium channels in hamster submandibular ganglion neurons

The Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are two novel neuropeptides which produce particular biological effects caused by interaction with G-protein-coupled receptors. We have shown in a previous study where VIP and PACAP 38 inhibit voltage-dependent calcium channel (VDCC) currents (ICa) via G-proteins in hamster submandibular ganglion (SMG) neurons. In this study, we attempt to further characterize the signal transduction pathways of VIP-and PACAP 38-induced modulation of ICa. Application of 1 microM VIP and PACAP 38 inhibited ICa by 33.0 +/- 3.1% and 36.8 +/- 2.6%, respectively (mean +/- S.E.M., n = 8). Application of strong voltage prepulse attenuated PACAP 38-induced inhibition of ICa. Pretreatment of cAMP dependent protein kinase (PKA) activator attenuated VIP-induced inhibition, but not the PACAP 38-induced inhibition. Intracellular dialysis of the PKA inhibitor attenuated the VIP-induced inhibition, but not the PACAP 38-induced inhibition. Pretreatment of protein kinase C (PKC) activator and inhibitor attenuated VIP-induced inhibition, but not the PACAP 38-induced inhibition. Pretreatment of cholera toxin (CTX) attenuated PACAP 38-induced inhibition of ICa. These findings indicate that there are multiple signaling pathways in VIP and PACAP 38-induced inhibitions of ICa: one pathway would be the VPAC1/VPAC2 receptors-induced inhibition involving both the PKA and PKC, and another one concerns the PAC1 receptor-induced inhibition via Gs-protein betagamma subunits. The VIP-and PACAP 38-induced facilitation of ICa can be observed in the SMG neurons in addition to inhibiting of ICa.