Onset of cholinergic efferent synaptic function in sensory hair cells of the rat cochlea.
In the developing mammalian cochlea, the sensory hair cells receive efferent innervation originating in the superior olivary complex. This input is mediated by α9/α10 nicotinic acetylcholine receptors (nAChRs) and is inhibitory due to the subsequent activation of calcium-dependent SK2 potassium channels. We examined the acquisition of this cholinergic efferent input using whole-cell voltage-clamp recordings from inner hair cells (IHCs) in acutely excised apical turns of the rat cochlea from embryonic day 21 to postnatal day 8 (P8). Responses to 1 mm acetylcholine (ACh) were detected from P0 on in almost every IHC. The ACh-activated current amplitude increased with age and demonstrated the same pharmacology as α9-containing nAChRs. Interestingly, at P0, the ACh response was not coupled to SK2 channels, so that the initial cholinergic response was excitatory and could trigger action potentials in IHCs. Coupling to SK current was detected earliest at P1 in a subset of IHCs and by P3 in every IHC studied. Clustered nAChRs and SK2 channels were found on IHCs from P1 on using Alexa Fluor 488 conjugated α-bungarotoxin and SK2 immunohistochemistry. The number of nAChRs clusters increased with age to 16 per IHC at P8. Cholinergic efferent synaptic currents first appeared in a subset of IHCs at P1 and by P3 in every IHC studied, contemporaneously with ACh-evoked SK currents, suggesting that SK2 channels may be necessary at onset of synaptic function. An analogous pattern of development was observed for the efferent synapses that form later (P6-P8) on outer hair cells in the basal cochlea.
Department of Otolaryngology, Head and Neck Surgery, The Center for Hearing and Balance, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Isabelle
I
Roux
Eric
E
Wersinger
J Michael
JM
McIntosh
Paul A
PA
Fuchs
Elisabeth
E
Glowatzki
D000109:Acetylcholine / Q000494:pharmacology; D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D001030:Apamin / Q000494:pharmacology; D001703:Biophysics; D002038:Bungarotoxins / Q000378:metabolism; D018678:Cholinergic Agents / Q000378:metabolism*; D003051:Cochlea / Q000166:cytology / Q000196:embryology / Q000254:growth & development; D004558:Electric Stimulation; D004622:Embryo, Mammalian; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018684:Glycine Agents / Q000494:pharmacology; D006198:Hair Cells, Auditory / Q000502:physiology*; D008297:Male; D018408:Patch-Clamp Techniques / Q000379:methods; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000378:metabolism; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000378:metabolism; D013331:Strychnine / Q000494:pharmacology; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
D002038:Bungarotoxins; D018678:Cholinergic Agents; C092350:FM 4-64; D018684:Glycine Agents; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D011978:Receptors, Nicotinic; D051640:Small-Conductance Calcium-Activated Potassium Channels; D001030:Apamin; D013331:Strychnine; D000109:Acetylcholine