Summary
The guanine nucleotide binding protein G0 alpha was immunolocalized in the guinea-pig vestibular system by confocal and electron microscopy. The vestibular sensory epithelia consist of the macula utriculi, macula sacculi and cristae ampullaris of the semicircular canals. Two types of hair cells are present in these epithelia. Type I hair cells are surrounded by an afferent nerve calyx that receives efferent innervation and type II hair cells are innervated directly by the afferent and efferent nerves. G0 alpha protein was observed on the inner face of the afferent calyceal membrane surrounding type I hair cells and in nerve endings in contact with type II hair cells. No labelling was found in the stereocilia and cuticular plate of type I and type II hair cells whereas the cytoplasmic matrix displayed a diffuse labelling. The plasma membrane of the supporting cells showed discreet labelling in the confocal microscope that are still confirmed by electron microscopy. A positive reaction was also observed along the plasma membrane of the vestibular ganglion neurons. Immunoblotting with affinity-purified polyclonal rabbit antibodies selective for the 39 kDa alpha subunit of G0 indicated that G0 alpha protein was present in both the vestibular ganglion. That G0 alpha labelling was observed in the cytoplasm of vestibular hair cells and in nerve endings contacting hair cells suggests that G0 may be involved in the modulation of vestibular neurotransmission.
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References
Anniko, M. &Arnold, W. (1991) Acetylcholine receptor localization in human adult cochlear and vestibular hair cells.Acta Oto-laryngologica 111, 491–9.
Annoni, J. M., Cochran, S. L. &Precht, W. (1984) Pharmacology of the vestibular hair cells afferent fibre synapse in the frog.Journal of Neuroscience 4, 2106–16.
Bartolami, S., Guiramand, J., Lenoir, M., Pujol, R. &Rêcasens, M. (1990) Carbachol-induced inositol phosphate formation during rat cochlea development.Hearing Research 47, 229–34.
Bartolami, S., Planche, M. &Pujol, R. (1993) Characterization of muscarinic binding sites in the adult and developing rat cochlea.Neurochemistry International 23, 419–25.
Birnbaumer, L., Abramowitz, J. &Brown, A. M. (1990) Receptor effector coupling by G proteins.Biochemica et Biophysica Acta 1031, 163–224.
Bockaert, J., Homburger, V. &Rouot, B. (1987) GTP binding proteins: a key role in cellular communication.Biochimie 69, 329–38.
Bockaert, J., Brabet, P., Gabrion, J., Homburger, V., Rouot, B., &Toutant, M. (1990) Structural, immunobiological, and functional characterization of guanine nucleotide-binding protein Go. InG Proteins (edited byIyengar, R. &Birnbaumer, L.) pp. 81–113. New York: Academic Press.
Brabet, P., Dumuis, A., Sebben, M., Pantaloni, C., Bockaert, J. &Homburger, V. (1988) Immunocytochemical localization of the guanine nucleotide-binding protein G0 in primary cultures of neuronal and glial cells.Journal of Neuroscience 8, 701–8.
Brabet, P., Homburger, V. &Bockaert, J. (1993) Neurobiology of G0. InHandbook of experimental pharmacology Vol. 108/11. GTAPases in biology II. (edited byDickey, B. F. &Birnbaumer, L.) pp. 429–46. New York: Springer-Verlag.
Brabet, P., Pantaloni, C., Rodriguez, M., Martinez, J., Bockaert, J. &Homburger, V. (1990) Neuroblastoma differentiation involves the expression of two isoforms of the alpha subunit of G0 Journal of Neurochemistry 54, 1310–20.
Canlon, B., Homburger, V. &Bockaert, J. (1991) The identification and localization of the guanine nucleotide binding protein G0 in the auditory system.European Journal of Neuroscience 3, 1338–42.
Demêmes, D., Raymond, J. &Sans, A. (1983) Selective retrograde labelling of vestibular efferent neurons with (3H) choline.Neuroscience 8, 285–90.
Demêmes, D., Wenthold, R. J., Moniot, B. &Sans, A. (1990) Glutamate-like immunoreactivity in the peripheral vestibular system of mammals.Hearing Research 46, 261–70.
Demêmes, D., Lleixa, A. &Dechesne, C. J. (1995) Cellular and subcellular localization of AMPA-selective glutamate receptors in the mammalian peripheral vestibular system.Brain Research 671, 83–94.
Dechesne, C. J., Raymond, J. &Sans, A. (1984) The action of glutamate in the cat labyrinth.Annals of Otology, Rhinology and Laryngology 93, 163–5.
Devau, G., Lehouelleur, J. &Sans, A. (1993) Glutamate receptors on type I vestibular hair cells of guinea-pigEuropean Journal of Neuroscience 5, 1210–17.
Dulon, D. &Schacht, J. (1992) Motility of cochlear outer hair cells.American Journal of Otology 13, 108–12.
Dulon, D., Moataz, R. &Mollard, P. (1993) Characterization of Ca2+ signals generated by extracellular nucleotides in supporting cells of the organ of Corti.Cell Calcium 14, 245–54.
Fesenko, E. E., Kolesnikov, S. S. &Lyubarsky, A. L. (1985) Induction by cyclic GMP of cationic conductance in plasma membrane of retinal rod outer segment.Nature 313, 310–13.
Gabrion, J., Brabet, P., Nguyen Than Dao, B., Homburger, V., Dumuis, A., Sebben, M., Rouot, B. &Bockaert, J. (1989) Ultrastructural localization of the GTP binding protein G0 in neurons.Cell Signalling 1, 107–32.
Griguer, C., Lehouelleur, J., Valat, J., Sahuquet, A. &Sans, A. (1993) Voltage dependent reversible movements of the apex in isolated guinea pig vestibular hair cells.Hearing Research 67, 110–16.
Guth, P. S., Norris, C. H. &Barron, S. E. (1988) Three tests of the hypothesis that glutamate is the sensory hair cells transmitter in the frog semicircular canal.Hearing Research 33, 223–8.
Guth, P. S., Aubert, A., Ricci, A. J. &Norris, C. H. (1991) Differential modulation of spontaneous and evoked neurotransmitter release from hair cell: some novel hypotheses.Hearing Research 56, 69–78.
Haynes, L. W. &Yau, K. W. (1985) Cyclic GMP-sensitive conductance in outer segment membranes of catfish cones.Nature 317, 61–3.
Homburger, V., Brabet, P., Audigier, Y., Pantaloni, C., Bockaert, J. &Rouot, B. (1987) Immunological localization of the GTP binding proteins G0 in different tissues of vertebrates and invertebrates.Molecular Pharmacology 31, 313–19.
Kakehata, S., Nakagawa, T., Akaike, N. &Takasaka, T. (1992) ACh activates inositol phosphate cascade via a PTX-sensitive G-protein in dissociated outer hair cells of guinea-pig cochlea.Proceedings Sendai Symposium 2, 25–9.
Kleuss, C., Hescheler, J., Ewel, C., Rosenthal, W., Schultz, G. &Wittig, B. (1991) Assignment of G-protein subtypes to specific receptors inducing inhibition of calcium currents.Nature 353, 43–8.
Neer, E. J., Lok, J. M. &Wolf, L. G. (1984) Purification and properties of the inhibitory guanine nucleotide regulatory unit of brain adenylate cyclase.Journal of Biological Chemistry 259, 14222–9.
Neer, E. &Clapham, D. E. (1988) Role of G proteins subunit in transmembrane signalling.Nature 333, 129–34.
Niedzielski, A. &Shacht, J. (1992) P2 purinoreceptors stimulate inositol phosphate release in the organ of Corti.Neuroreport 3, 273–5.
Ogawa, K. &Schacht, J. (1993) Receptor-mediated release of inositol phosphates in the cochlear and vestibular epithelia of the rat.Hearing Research 69, 207–14.
Ohtani, M., Devau, G., Lehouelleur, J. &Sans, A. (1994) Cholinergic agonists increase intracellular calcium concentration in frog vestibular hair cells.Hearing Research 80, 167–73.
Pace, U. &Lancet, D. (1986) Olfactory GTP-binding protein: signal-transducing polypeptide of vertebrate chemosensory neurons.Proceedings of the National Academy of Sciences (USA) 83, 4947–95.
Prigioni, I., Russo, G., Valli, P. &Masetto, S. (1990) Pre- and postsynaptic excitatory action of glutamate agonist on frog vestibular receptors.Hearing Research 46, 253–60.
Rouot, B., Charpentier, N., Chabbert, C., Carrette, J., Zumbihl, R., Bockaert, J. &Homburger, V. (1992) Specific antibodies G0 isoforms reveal the early expression of G02 alpha subunit and appearance of G01 alpha during neuronal differentiation.Molecular Pharmacology 41, 273–80.
Schwartz, D. W. F., Satoh, K., Schwartz, I. E., Hu, K. &Fibiger, H. C. (1986) Cholinergic innervation of the rat's labryinth.Experimental Brain Research 64, 19–26.
Shigemoto, T. &Ohmori, H. (1990) Muscarinic agonists and ATP increase the intracellular calcium concentration in chick cochlear hair cells.Journal of Physiology 420, 127–48.
Slepecky, N. &Ulfendahl, M. (1988) Glutaraldehyde induces cell shape changes in isolated outer hair cells from the inner ear.Journal of Submicroscopy Cytology and Pathology 20, 37–45.
Sternweis, P. C. &Robishaw, J. D. (1984) Isolation of two proteins with high affinity for guanine nucleotides for membranes from bovine brain.Journal of Biological Chemistry 259, 13806–13.
Tonosaki, K. &Funakoshi, M. (1988) Cyclic nucleotides may mediate taste transduction.Nature 331, 354–6.
Van Megen, Y. J. B., Klaasen, A. M. B., Rodrigues De Miranda, J. F. &Kuijpers, W. (1988) Cholinergic muscarinic receptors in rat cochlea.Brain Research 474, 185–8.
Valat, J., Devau, G., Dulon, D. &Sans, A. (1989) Vestibular hair cells isolated from guinea-pig labryinth.Hearing Research 40, 255–60.
Valat, J., Griguer, C., Lehouelleur, J. &Sans, A. (1991) Motile responses of isolated guinea-pig vestibular hair cells.Neuroscience Letters 127, 231–6.
Yoshida, N., Shigemoto, T., Sugai, T. &Ohmori, H. (1994) The role of inositol triphosphate on Ach-induced outward currents in bullfrog saccular hair cells.Brain Research 644, 90–100.
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Valat, J., Scarfone, E., Travo, C. et al. Immunocytochemical localization of the GTP-binding protein G0 alpha in the vestibular epithelium and ganglion of the guinea-pig. J Neurocytol 24, 916–924 (1995). https://doi.org/10.1007/BF01215642
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DOI: https://doi.org/10.1007/BF01215642