Muscarinic acetylcholine receptor subtype expression in type vestibular hair cells of guinea pigs

Abstract

Recent studies have demonstrated that five subtypes (M1-M5) of muscarinic acetylcholine receptor (mAChR) are expressed in the vestibular periphery. However, the exact cellular location of the mAChRs is not clear. In this study, we investigated whether there is the expression of M1-M5 muscarinic receptor mRNA in isolated type II vestibular hair cells of guinea pig by using single-cell RT-PCR. In vestibular end-organ, cDNA of the expected size was obtained by RT-PCR. Moreover, mRNA was identified by RT-PCR from individually isolated type II vestibular hair cells (single-cell RT-PCR). Sequence analysis confirmed that the products were M1-M5 mAChR. These results demonstrated that M1-M5 mAChR was expressed in the type II vestibular hair cells of the guinea pig, which lends further support for the role of M1-M5 mAChR as a mediator of efferent cholinergic signalling pathway in vestibular hair cells.

Keywords

muscarinic acetylcholine receptor / type II vestibular hair cells / single-cell RT-PCR

References

[1] EybalinM.. Neurotransmitters and neuromodulators of the mammalian cochlea. Physiol Rev, 1993, 73(2): 309-373

[2] GuthP.S., DunnA., KronomerK., et al.. The cholinergic pharmacology of the frog saccule. Hear Res, 1994, 75(1–2): 225-232

[3] JaggerD.J., GriesingerC.B., RivoltaM.N., Holley, et al.. Calcium signaling mediated by the α9 acetylcholine recetor in a cochlear cell line from the immortomouse. J Physiol, 2000, 527(Pt1): 49-54

[4] IshiyamaA.I., LopezI., WackymP.A.. Choline acetyltransferase immunoreactivity in the human vestibular end-organs. Cell Biol Int, 1994, 18(10): 979-984

[5] MatsudaY.. Localization of choline acetyltransferase and calcitonin gene-related peptide immunoreactivities in the vestibular end-organs of the guinea pig. Osaka City Med J, 1996, 42(2): 61-76

[6] KongW.J., EggG., HusslB., et al.. Localization of chat-like immunoreactivity in the vestibular endorgans of the rat. Hear Res, 1994, 75(1–2): 191-200

[7] KongW.J., ScholtzA.W., HusslB., et al.. Localization of eVerent neurotransmitters in the inner ear of the homozygous Bronx waltzer mutant mouse. Hear Res, 2002, 167(1–2): 136-155

[8] KongW.J., HusslB., ThumfartW.F., et al.. Ultrastructural localization of ChAT-like immunoreactivity in the human vestibular periphery. Hear, 1998, 119(1–2): 96-103

[9] PopperP., IshiyamaA., LopezI., et al.. Calcitonin gene-related peptide and choline acetyltransferase immunoreactivity in the human vestibular periphery. Audiol Neurootol, 2002, 7(5): 298-302

[10] WackymP.A., ChenC.T., IshiyamaA., et al.. Muscarinic acetylcholine receptor subtype mRNAs in the human and rat vestibular periphery. Cell Biol Int, 1996, 20(3): 187-192

[11] HielH., ElgoyhenA.B., DrescherD.G., et al.. Expression of nicotinic acetylcholine receptor mRNA in the adult rat peripheral vestibular system. Brain Res, 1996, 738(2): 347-352

[12] AndersonA.D., TroyanovskayaM., WackymP.A.. Differential expression of K2-9 and L2-4 nicotinic acetylcholine receptor subunit mRNA in the vestibular end-organs and Scarpa’s ganglia of the rat. Brain Res, 1997, 778(2): 409-413

[13] IshiyamaA.I., LopezI., WackymP.A.. Molecular characterization of muscarinic receptors in the human vestibular periphery. Implications for pharmacotherapy. Am J Otol, 1997, 18(5): 648-654

[14] ElgoyhenA.B., VetterD.E., KatzE., et al.. α10: A determinant of nicotinic cholinergic receptor function in mammalian vestibular and cochlear mechanosensory hair cells. Proc Natl Acad Sci USA, 2001, 98(6): 3501-3506

[15] LiG.Q., KevetterG.A., LeonardR.B., et al.. Muscarinic acetylcholine receptor subtype expression in avian vestibular hair cells, nerve terminals and ganglion cells. Neuroscience, 2007, 146(1): 384-402

[16] KhanK.M., DrescherM.J., HatfieldJ.S., et al.. Muscarinic receptor subtypes are differentially distributed in the rat cochlea. Neuroscience, 2002, 111(2): 291-302

[17] CaulfieldM.P.. Muscarinic receptors—characterization, coupling and function. Pharmacol Ther, 1993, 58(3): 319-379

[18] EglenR.M., NahorskiS.R.. The muscarinic M(5) receptor: a silent or emerging subtype?. Br J Pharmacol, 2000, 130(1): 13-21

[19] GuthP.S., PerinP., NorrisC.H., et al.. The vestibular hair cells: post-transductional signal processing. Prog Neurobiol, 1998, 54(2): 193-247

[20] TsengJ., ErbeC.B., KwitekA.E., et al.. Radiation hybrid mapping of five muscarinic acetylcholine receptor subtype genes in Rattus norvegicus. Hear Res, 2002, 174(1–2): 86-92

[21] KongW.J., GuoC.K., ZhangS., et al.. The properties of ACh-induced BK currents in guinea pig type II vestibular hair cells. Hear Res, 2005, 209(1-2): 1-9

[22] SugaiT., SugitaniM., OoyamaH.. Effects of activation of the divergent efferent fibers on the spontaneous activity of vestibular afferent fibers in the toad. Jpn J Physiol, 1991, 41(2): 217-232

[23] KakehataS., NakagawaT., TakasakaT., et al.. Cellular mechanism of ACh-induced response in dissociated outer hair cells of guinea pig cochlea. J Physiol, 1993, 463: 227-244

[24] LiuQ., WuJ., WangX., et al.. Changes in muscarinic acetylcholine receptor expression in form deprivation myopia in guinea pigs. Mol Vis, 2007, 13: 1234-1244

[25] RicciA., RennieK., CochranS., et al.. Vestibular type I and type II hair cells. 1: Morphometric identification in the pigeon and gerbil. J Vestibular Res, 1997, 7(5): 393-406

[26] NorrisC.H., HousleyG.D., WilliamsW.H., et al.. The acetylcholine receptors of the semicircular canal in the frog (Rana pipiens). Hear, 1988, 32(2–3): 197-206

[27] SridharT.S., LibermanM.C., BrownM.C., et al.. Anovel cholinergic “slow eVect” of eVerent stimulation on cochlear potentials in guinea pig. J Neurosci, 1995, 15(5Pt1): 3667-3678

[28] HoltJ.C., LioudynoM., AthasG., et al.. The effect of proteolytic enzymes on the alpha9-nicotinic receptormediated response in isolated frog vestibular hair cells. Hear Res, 2001, 152(1–2): 25-42

[29] SugaiT., YanoJ., SugitaniM., et al.. Actions of cholinergic agonists and antagonists on the efferent synapse in the frog sacculus. Hear, 1992, 61(1–2): 56-64

[30] ElgoyhenA.B., JohnsonD.S., BoulterJ., et al.. α9: An acetylcholine receptor with novel pharmacological properties expressed in rat cochlear hair cells. Cell, 1994, 79(4): 705-715

[31] GlowatzkiE., FuchsP.A.. Cholinergic synaptic inhibition of inner hair cells in the neonatal mammalian cochlea. Science, 2000, 288(5475): 2366-2368

[32] KatzE., VerbitskyM., RothlinC.V., et al.. High calcium permeability and calcium block of the alpha9 nicotinic acetylcholine receptor. Hear Res, 2000, 141(1–2): 117-128

[33] OliverD., KlockerN., SchuckJ., et al.. Gating of Ca²⁺-activated K⁺ channels controls fast inhibitory synaptic transmission at auditory outer hair cells. Neuron, 2000, 26(3): 595-601

[34] FuchsP.A., MurrowB.W.. Cholinergic inhibition of short (outer) hair cells of the chicks cochlea. J Neurosci, 1992, 12(3): 800-809

[35] NenovA.P., NorrisC.H., BobbinR.P.. Acetylcholine response in guinea pig outer hair cells. I. Properties of the response. Hear Res, 1996, 101(1–2): 132-148

[36] KongW.J., ChengH.M., van CauwenbergeP.. Expression of nicotinic acetylcholine receptor subunit alpha9 in type II vestibular hair cells of rats. Acta Pharmacol Sin, 2006, 27(11): 1509-1514

[37] VetterD.E., LibermanM.C., MannJ., et al.. Role of alpha9 ACh receptor subunits in the development and function of cochlear eVerent innervation. Neuron, 1999, 23(1): 93-103

[38] FelderC.C.. Muscarinic acetylcholine receptors: signal transduction through multiple eVectors. FASEB J, 1995, 9(8): 619-625