Vestibular hair cells of the chick express the nicotinic acetylcholine receptor subunit alpha 9.

The efferent cholinergic pathways to the vestibular periphery have yet to be fully characterized. While the nicotinic acetylcholine receptor subunit (nAChR) alpha 9 is now regarded as the principle receptor for efferent cholinergic signaling to the organ of Corti, there is still uncertainty over how the more complex efferent effects of the labyrinth are produced. Recent experimental work has demonstrated that the nAChR alpha 9 is present in the vestibular end-organs of the rat and mouse, suggesting that alpha 9 may be one of the mediators of efferent cholinergic signaling in the vestibular periphery as well. In this experiment, we sought to determine whether alpha 9 was also present in the vestibular end-organs of the chick. A homologue of alpha 9 has been cloned recently from the chick cochlea. Using reverse transcription polymerase chain reaction (RT-PCR), individual vestibular end-organ preparations, including posterior ampulla, combined horizontal and superior ampulla, saccule, utricle, and the vestibular ganglion were screened for alpha 9 messenger RNA expression. In each end-organ and the vestibular ganglion, a cDNA of the expected size was obtained by RT-PCR and was confirmed to be alpha 9 by sequence analysis. Further, alpha 9 mRNA was identified by RT-PCR from individually isolated type I and type II vestibular hair cells (single-cell RT-PCR). Lastly, insitu hybridization using digoxigenin-labeled alpha 9 riboprobes confirmed the presence of alpha 9 in type I and type II hair cells throughout the vestibular periphery. These results demonstrate the expression of alpha 9 in the vestibular end-organs of the chick, and lend further support for the role of alpha 9 as a mediator of efferent cholinergic signaling in vestibular hair cells.