title: AMPA receptor-mediated rapid EPSCs in vestibular calyx afferents.
abstract: In the vestibular periphery neurotransmission between hair cells and primary afferent nerves occurs via specialized ribbon synapses. Type I vestibular hair cells (HCIs) make synaptic contacts with calyx terminals, which enclose most of the HCI basolateral surface. To probe synaptic transmission, whole cell patch-clamp recordings were made from calyx afferent terminals isolated together with their mature HCIs from gerbil crista. Neurotransmitter release was measured as excitatory postsynaptic currents (EPSCs) in voltage clamp. Spontaneous EPSCs were classified as simple or complex. Simple events exhibited a rapid rise time and a fast monoexponential decay (time constant < 1 ms). The remaining events, constituting ~40% of EPSCs, showed more complex characteristics. Extracellular Sr2+greatly increased EPSC frequency, and EPSCs were blocked by the AMPA receptor blocker NBQX. The role of presynaptic Ca2+channels was assessed by application of the L-type Ca2+channel blocker nifedipine (20 µM), which reduced EPSC frequency. In contrast, the L-type Ca2+channel opener BAY K 8644 increased EPSC frequency. Cyclothiazide increased the decay time constant of averaged simple EPSCs by approximately twofold. The low-affinity AMPA receptor antagonist γ-d-glutamylglycine (2 mM) reduced the proportion of simple EPSCs relative to complex events, indicating glutamate accumulation in the restricted cleft between HCI and calyx. In crista slices EPSC frequency was greater in central compared with peripheral calyces, which may be due to greater numbers of presynaptic ribbons in central hair cells. Our data support a role for L-type Ca2+channels in spontaneous release and demonstrate regional variations in AMPA-mediated quantal transmission at the calyx synapse.NEW & NOTEWORTHYIn vestibular calyx terminals of mature cristae we find that the majority of excitatory postsynaptic currents (EPSCs) are rapid monophasic events mediated by AMPA receptors. Spontaneous EPSCs are reduced by an L-type Ca2+channel blocker and notably enhanced in extracellular Sr2+EPSC frequency is greater in central areas of the crista compared with peripheral areas and may be associated with more numerous presynaptic ribbons in central hair cells.
authors.affiliation: Department of Otolaryngology, University of Colorado School of Medicine, Aurora, Colorado.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Kirk
authors.affiliation: Department of Otolaryngology, University of Colorado School of Medicine, Aurora, Colorado.
authors.forename: Frances L
authors.initials: FL
authors.lastname: Meredith
authors.affiliation: Department of Otolaryngology, University of Colorado School of Medicine, Aurora, Colorado.|Departments of Pediatrics, Neurology, and Pharmacology, University of Colorado School of Medicine, Aurora, Colorado.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Benke
authors.affiliation: Department of Otolaryngology, University of Colorado School of Medicine, Aurora, Colorado; katie.rennie@ucdenver.edu.|Department of Physiology and Biophysics, University of Colorado School of Medicine, Aurora, Colorado; and.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Rennie
mesh_terms: D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D001581:Benzothiadiazines / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000378:metabolism; D002478:Cells, Cultured; D004151:Dipeptides / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials; D005260:Female; D005849:Gerbillinae; D018069:Hair Cells, Vestibular / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D008297:Male; D009543:Nifedipine / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D018091:Receptors, AMPA / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013324:Strontium / Q000494:pharmacology; D013569:Synapses / Q000187:drug effects / Q000378:metabolism / Q000502:physiology
chemical_list: D001581:Benzothiadiazines; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D004151:Dipeptides; D011810:Quinoxalines; D018091:Receptors, AMPA; C062865:2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline; C028929:gamma-glutamylglycine; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D009543:Nifedipine; C004639:cyclothiazide; D013324:Strontium
keywords: AMPA receptor; crista; cyclothiazide; glutamate; type I hair cell