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The authors use 3-D tomographic reconstruction to provide a detailed description of the components of the hair cell active zone, including the ribbon, tethered and distant vesicles, and other membranous organelles. Following prolonged stimulation, vesicles docked at the plasma membrane were depleted regardless of their distance from the ribbon. Ribbon-tethered vesicles were preferentially lost from the side of the synaptic body nearer the plasma membrane.
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The authors used in situ hybridization to show that cysteine string protein (CSP) mRNA was present in inner, but not outer hair cells of the rat cochlea. CSP was also expressed in spiral ganglion neurons. Antibodies to CSP co-localized with antibodies to syntaxin and SNAP-25 in inner hair cells, and in efferent nerve terminals that innervate outer hair cells. Immunoreactivity could also be detected by electron microscopy to be associated with synaptic vesicles and efferent nerve terminals.
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The authors used flash photolysis of caged calcium and capacitance measurements to determine the calcium dependence of exocytosis and endocytosis in hair cells of the mouse cochlea. Rapid elevation of calcium above 8 μM caused a biphasic capacitance increase corresponding to the fusion of around 40,000 vesicles. This is many times greater than the 300 or so vesicles thought to be docked immediately at the active zones, and so must include fusion-competent vesicles docked some distance from the active zone.
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The authors combined single unit recording and cochlear perfusion in vivo to demonstrate that dihydropyridine-sensitive calcium channels support afferent synaptic transmission in the mammalian cochlea.
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The authors used whole-cell voltage clamp recording to describe the progressive acquisition of different types of voltage-gated potassium currents by inner hair cells in the mouse cochlea. These changes convert the hair cell from a pattern of calcium electrogenesis to graded depolarizations produced by sound.
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in press. Published online. The authors perfused specific toxins into the guinea pig cochlea to examine the role of BK channels (calcium-sensitive, voltage-gated potassium channels) in synaptic transmission in vivo. Charybdotoxin or iberiotoxin reversibly blocked the compound action potential but had no effect on signals generated by outer hair cells. As BK channels appear to be expressed by inner hair cells and by the afferent neurons, the effects of toxin on sound evoked afferent signals could occur presynaptically, postsynaptically, or both
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+ channels to auditory neurotransmission in the guinea pig cochlea. J Neurophysiol 2003, in press. Published online: http://jn.physiology.org/cgi/reprint/01155.2002v1.pdf The authors perfused specific toxins into the guinea pig cochlea to examine the role of BK channels (calcium-sensitive, voltage-gated potassium channels) in synaptic transmission in vivo. Charybdotoxin or iberiotoxin reversibly blocked the compound action potential but had no effect on signals generated by outer hair cells. As BK channels appear to be expressed by inner hair cells and by the afferent neurons, the effects of toxin on sound evoked afferent signals could occur presynaptically, postsynaptically, or both.
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The authors used measurements of voltage-gated calcium current and membrane capacitance to chart the maturation of synaptic function in hair cells of the mouse cochlea. Calcium-dependent exocytosis and endocytosis were operational at least one week before the onset of hearing, around postnatal day 10. Calcium current density and capacitance changes reached a peak about postnatal day 6, then fell to adult levels by postnatal day 14. Although smaller at maturity, exocytosis was more efficiently coupled to calcium influx in the mature hair cells, perhaps reflecting structural changes seen in synaptic ribbons over this time course.
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The authors recorded excitatory postsynaptic currents (EPSCs) from afferent boutons at their point of contact with inner hair cells in an excised preparation of the neonatal rat cochlea. EPSCs occurred infrequently at rest (around 1 Hz) but could be raised in frequency by bath application of high potassium saline. EPSCs were rapid and of variable amplitude. Both interval and amplitude histograms suggested that larger EPSCs occurred through the coordinate release of multiple vesicles of neurotransmitter.
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Glowatzki E., Fuchs P.A. Transmitter release at the hair cell ribbon synapse. Nat. Neurosci. 5:2002;147-154 The authors recorded excitatory postsynaptic currents (EPSCs) from afferent boutons at their point of contact with inner hair cells in an excised preparation of the neonatal rat cochlea. EPSCs occurred infrequently at rest (around 1 Hz) but could be raised in frequency by bath application of high potassium saline. EPSCs were rapid and of variable amplitude. Both interval and amplitude histograms suggested that larger EPSCs occurred through the coordinate release of multiple vesicles of neurotransmitter.
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Distribution of the glutamate/aspartate transporter GLAST in relation to the afferent synapses of outer hair cells in the guinea pig cochlea
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The authors used RT-PCR cloning, in situ hybridization and immunohistology to map the expression of various glutamate transporters in the guinea-pig cochlea. In addition to GLAST expression in supporting cells, GLT-1 (glutamate transporter-1) and EAAC1 were detected in spiral ganglion neurons, the cochlear afferents.
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Goldfish retinal bipolar cells and frog saccular hair cells were studied using evanescent wave fluorescence microscopy to examine near-membrane, voltage-gated calcium influx. At discrete locations calcium rose and fell contemporaneously with the opening and closing of voltage-gated calcium channels. These calcium 'hotspots' were found to correspond in density to those of synaptic ribbons labeled with antibodies to the protein 'Ribeye'.
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