Constitutive expression of the alpha10 nicotinic acetylcholine receptor subunit fails to maintain cholinergic responses in inner hair cells after the onset of hearing.
Efferent inhibition of cochlear hair cells is mediated by alpha9alpha10 nicotinic cholinergic receptors (nAChRs) functionally coupled to calcium-activated, small conductance (SK2) potassium channels. Before the onset of hearing, efferent fibers transiently make functional cholinergic synapses with inner hair cells (IHCs). The retraction of these fibers after the onset of hearing correlates with the cessation of transcription of the Chrna10 (but not the Chrna9) gene in IHCs. To further analyze this developmental change, we generated a transgenic mice whose IHCs constitutively express alpha10 into adulthood by expressing the alpha10 cDNA under the control of the Pou4f3 gene promoter. In situ hybridization showed that the alpha10 mRNA is expressed in IHCs of 8-week-old transgenic mice, but not in wild-type mice. Moreover, this mRNA is translated into a functional protein, since IHCs from P8-P10 alpha10 transgenic mice backcrossed to a Chrna10(-/-) background (whose IHCs have no cholinergic function) displayed normal synaptic and acetylcholine (ACh)-evoked currents in patch-clamp recordings. Thus, the alpha10 transgene restored nAChR function. However, in the alpha10 transgenic mice, no synaptic or ACh-evoked currents were observed in P16-18 IHCs, indicating developmental down-regulation of functional nAChRs after the onset of hearing, as normally observed in wild-type mice. The lack of functional ACh currents correlated with the lack of SK2 currents. These results indicate that multiple features of the efferent postsynaptic complex to IHCs, in addition to the nAChR subunits, are down-regulated in synchrony after the onset of hearing, leading to lack of responses to ACh.
Taranda
Julián
J
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, 1428, Argentina.
Ballestero
Jimena A
JA
Hiel
Hakim
H
de Souza
Flavio S J
FS
Wedemeyer
Carolina
C
Gómez-Casati
M Eugenia
ME
Lipovsek
Marcela
M
Vetter
Douglas E
DE
Fuchs
Paul A
PA
Katz
Eleonora
E
Elgoyhen
A Belén
AB
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D018678:Cholinergic Agents / Q000494:pharmacology; D006199:Hair Cells, Auditory, Inner / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006309:Hearing / Q000187:drug effects / Q000502:physiology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000378:metabolism*; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000187:drug effects / Q000378:metabolism; D050821:Transcription Factor Brn-3C / Q000235:genetics / Q000378:metabolism
D018678:Cholinergic Agents; C526014:Chrna10 protein, mouse; D018398:Homeodomain Proteins; C496898:Kcnn2 protein, mouse; C121290:Pou4f3 protein, mouse; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D051640:Small-Conductance Calcium-Activated Potassium Channels; D050821:Transcription Factor Brn-3C; D000109:Acetylcholine