Skip to main content
Log in

Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate

  • Published:
Experimental Brain Research Aims and scope Submit manuscript

Summary

The light- and electron microscopic localization of glutamate and glutamine in the rat organ of Corti was studied by means of antisera raised against the respective amino acids coupled to carrier proteins. The light microscopic analysis was performed in semithin sections treated according to the peroxidase-antiperoxidase procedure. The two amino acids were visualized in the same ultrathin sections by use of postembedding immunocytochemistry with two different gold particle sizes. The distribution of aspartate-like immunoreactivity was also recorded, but only at the light microscopic level. In the hair cells, the level of glutamate-like immunoreactivity was higher than that in supporting cells but lower than that in the presumed glutamatergic terminals of cerebellar parallel and mossy fibres. The latter types of terminal were sampled from ultrathin sections that had been incubated under the same conditions as the cochlear sections. Within the hair cells, gold particles signalling glutamate were enriched on mitochondria but not on clusters of synaptic vesicles. Glutamine-like immunoreactivity was present in hair cells as well as supporting cells. The glutamate/glutamine ratio, expressed as the ratio between the respective gold particle densities, was considerably lower for hair cells compared with the cerebellar excitatory terminals. No consistent difference was found between outer and inner hair cells in relation to the levels and subcellular distribution of glutamate and glutamine immunoreactivities. Aspartate-like immunoreactivity was accumulated in outer hair cells, with some labelling also of border cells and Böttcher cells. While the present study confirmed the presence of glutamate in hair cells and demonstrated that these cells are also endowed with the important glutamate precursor glutamine, it revealed notable differences between hair cells and presumed glutamatergic terminals in the CNS. These could reflect differences in the synthesis and compartmentation of transmitter glutamate. Methodological factors could also contribute. Alternatively, the differences could be interpreted to suggest that the hair cell transmitter is not glutamate, but a similar compound. Aspartate could be a candidate in the case of the outer hair cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Altschuler RA, Sheridan CE, Horn JW, Wenthold RJ (1989) Immunocytochemical localization of glutamate immunoreactivity in the guinea pig cochlea. Hear Res 42:167–174

    Google Scholar 

  • Anniko M, Lundquist P-G (1980) Temporal bone morphology after systemic arterial perfusion or intralabyrinthine in situ immersion: 1. Hair cells of the vestibular organs and the cochlea. Micron 11:73–83

    Google Scholar 

  • Bendayan M, Nanci A, Kan FWK (1987) Effect of tissue processing on colloidal gold cytochemistry. J Histochem Cytochem 35:983–996

    Google Scholar 

  • Blackstad TW, Karagülle T, Ottersen OP (1990) MORFOREL, a computer program for two-dimensional analysis of micrographs of biological specimens, with emphasis on immunogold preparations. Comput Biol Med 20:15–34

    Google Scholar 

  • Bledsoe SC Jr, Bobbin RP, Puel J-L (1988) Neurotransmission in the inner ear. In: Jahn AF, Santos-Sacchi J (eds) Physiology of the ear. Raven Press, New York, pp 385–406

    Google Scholar 

  • Bobbin RP (1979) Glutamate and aspartate mimic the afferent transmitter in the cochlea. Exp Brain Res 34:389–393

    Google Scholar 

  • Bobbin RP, Ceasar G, Fallon M (1990) Potassium induced release of GABA and other substances from the guinea pig cochlea. Hear Res 46:83–94

    Google Scholar 

  • Bobbin RP, Ceasar G, Fallon M (1991) Changing cation levels (Mg2+, Ca2+, Na+) alters the release of glutamate, GABA and other substances from the guinea pig cochlea. Hear Res 54:135–144

    Google Scholar 

  • Bramham CR, Torp R, Zhang N, Storm-Mathisen J, Ottersen OP (1990) Distribution of glutamate-like immunoreactivity in excitatory hippocampal pathways: a semiquantitative electron microscopic study in rats. Neuroscience 39:405–417

    Google Scholar 

  • Burger PM, Mehl E, Cameron PL, Maycox PR, Baumbert M, Lottspeich F, De Camilli P, Jahn R (1989) Synaptic vesicles immuno-isolated from rat cerebral cortex contain high levels of glutamate. Neuron 3:715–720

    Google Scholar 

  • Cotman CW, Monaghan DT, Ottersen OP, Storm-Mathisen J (1987) Anatomical organization of excitatory amino acid receptors and their pathways. Trends in Neurosci 10:273–280

    Google Scholar 

  • Dale N, Ottersen OP, Roberts A, Storm-Mathisen J (1986) Inhibitory neurones of a motor pattern generator in Xenopus revealed by antibodies to glycine. Nature 324:255–257

    Google Scholar 

  • Do KQ, Mattenberger M, Streit P, Cuénod M (1986) In vitro release of endogenous excitatory sulfur-containing amino acids from various rat brain regions. J Neurochem 46:779–786

    Google Scholar 

  • Drescher MJ, Drescher DG, Medina JE (1983) Effect of sound stimulation at several levels on concentrations of primary amines, including neurotransmitter candidates, in perilymph of the guinea pig inner ear. J Neurochem 41:309–320

    Google Scholar 

  • Ehrenberger K, Felix D (1991) Glutamate receptors in afferent cohclear neurotransmission in guinea pigs. Hear Res 52:73–80

    Google Scholar 

  • Erecinska M, Silver IA (1990) Metabolism and role of glutamate in mammalian brain. Prog Neurobiol 35:245–296

    Google Scholar 

  • Eybalin M, Pujol R (1983) A radioautographic study of [3H]L glutamate and [3H]L-glutamine uptake in the guinea-pig cochlea. Neuroscience 9:863–871

    Google Scholar 

  • Eybalin M, Renard N, Ottersen OP, Storm-Mathisen J, Pujol R (1991) Ultrastructural immunocalization of glutamate in the guinea pig organ of Corti. Association for Research in Otolaryngology, Abstracts of the 14th Midwinter Research Meeting, p 18

  • Fex J, Altschuler RA (1986) Neurotransmitter-related immunocytochemistry of the organ of Corti. Hear Res 22:249–263

    Google Scholar 

  • Fonnum F (1984) Glutamate: a transmitter in mammalian brain. J Neurochem 42:1–11

    Google Scholar 

  • Gleich O, Johnstone BM, Robertson D (1990) Effects of l-glutamate on auditory afferent activity in view of its proposed excitatory transmitter role in the mammalian cochlea. Hear Res 45:295–312

    Google Scholar 

  • Godfrey DA, Wiet GJ, Ross CD (1986) Quantitative histochemistry of the cochlea. In: Altschuler RA, Hoffman DW, Bobbin RP (eds) Neurobiology of hearing: the cochlea. Raven Press, New York, pp 149–160

    Google Scholar 

  • Godfrey DA, Carter JA, Berger SJ, Matschinsky FM (1976) Levels of putative transmitter amino acids in the guinea pig cochlea. J Histochem Cytochem 24:468–472

    Google Scholar 

  • Griffiths R (1990) Cysteine sulphinate (CSA) as an excitatory amino acid transmitter candidate in the mammalian central nervous system. Prog Neurobiol 35:313–323

    Google Scholar 

  • Jenison GL, Bobbin RP, Thalmann R (1985) Potassium-induced release of endogenous amino acids in the guinea pig cochlea. J Neurochem 44:1845–1853

    Google Scholar 

  • Ji Z, Aas J-E, Laake J, Walberg F, Ottersen OP (1991) An electron microscopic immunogold analysis of glutamate and glutamine in terminals of rat spinocerebellar fibers. J Comp Neurol 307:296–310

    Google Scholar 

  • Klinke R (1986) Neurotransmission in the inner ear. Hear Res 22:235–243

    Google Scholar 

  • Kvamme E (1983) Glutamine. In: Lajtha A (eds) Handbook of neurochemistry. Plenum Press, New York, pp 405–489

    Google Scholar 

  • Laake JH, Gundersen V, Nordbø G, Ottersen OP, Storm-Mathisen J (1986) An antiserum against glutamine. In: Roberts PJ, Storm-Mathisen J, Bradford HF (eds) Excitatory amino acids. Macmillan, London, pp 448–450

    Google Scholar 

  • Madsen S, Ottersen OP, Storm-Mathisen J, Sturman JA (1990) Immunocytochemical localization of taurine: methodological aspects. In: Pasantes-Morales H et al (eds) Taurine: Functional neurochemistry, physiology, and cardiology. Wiley-Liss, New York, pp 37–44

    Google Scholar 

  • Melamed B, Norris B, Bryant G, Guth P (1982) Amino acid contents of guinea pig perilymph collected under conditions of quiet or sound stimulation. Hear Res 7:13–18

    Google Scholar 

  • Montero VM, Wenthold RJ (1989) Quantitative immunogold analysis reveals high glutamate levels in retinal and cortical synaptic terminals in the lateral geniculate nucleus of the macaque. Neuroscience 31:639–647

    Google Scholar 

  • Ottersen OP (1987) Postembedding light- and electron microscopic immunocytochemistry of amino acids: description of a new model system allowing identical conditions for specificity testing and tissue processing. Exp Brain Res 69:167–174

    Google Scholar 

  • Ottersen OP (1989) Quantitative electron microscopic immunocytochemistry of amino acids. Anat Embryol 180:1–15

    Google Scholar 

  • Ottersen OP, Zhang N, Walberg F (1992) Metabolic compartmentation of glutamate and glutamine: morphological evidence obtained by quantitative immunocytochemistry in rat cerebellum. Neuroscience 46:519–534

    Article  CAS  PubMed  Google Scholar 

  • Palaiologos G, Hertz L, Schousboe A (1989) Role of aspartate aminotransferase and mitochondrial dicarboxylate transport for release of endogenously and exogenously supplied neurotransmitter in glutamatergic neurons. Neurochem Res 14:359–366

    Google Scholar 

  • Puel J-L, Ladrech S, Chabert R, Pujol R, Eybalin M (1991a) Electrophysiological evidence for the presence of NMDA receptors in the guinea pig cochlea. Hear Res 51:255–264

    Google Scholar 

  • Puel J-L, Pujol R, Ladrech S, Eybalin M (1991b) α-amino-3 hydroxy-5-methyl-4-isoxazole propionic acid electrophysiological and neurotoxic effects in the guinea-pig cochlea. Neuroscience 45:63–72

    Google Scholar 

  • Pujol R, Lenoir M, Robertson D, Eybalin M, Johnstone BM (1985) Kainic acid selectively alters auditory dendrites connected with cochlear inner hair cells. Hear Res 18:145–151

    Google Scholar 

  • Pujol R, Rebillard G, Puel J-L, Lenoir M, Eybalin M, Recasens M (1991) Glutamate neurotoxicity in the cochlea: a possible consequence of ischaemic or anoxic conditions occurring in ageing. Acta Otolaryngol Suppl (Stockh) 476:32–36

    Google Scholar 

  • Ryan AF, Schwartz IR (1984) Preferential glutamine uptake by cochlear hair cells: implications for the afferent cochlear transmitter. Brain Res 290:376–379

    Google Scholar 

  • Sewell WF, Mroz EA (1990) Purification of a low-molecular-weight excitatory substance from the inner ears of goldfish. Hear Res 50:127–138

    Google Scholar 

  • Somogyi P, Halasy K, Somogyi J, Storm-Mathisen J, Ottersen OP (1986) Quantification of immunogold labelling reveals enrichment of glutamate in mossy and parallel fibre terminals in cat cerebellum. Neuroscience 19:1045–1050

    Google Scholar 

  • Somogyi P, Hodgson AJ, Smith AD, Nunzi MG, Gorio A, Wu J-Y (1984) Different populations of GABAergic neurons in the visual cortex and hippocampus of cat contain somatostatinor cholecystokinin-immunoreactive material. J Neurosci 4:2590–2603

    Google Scholar 

  • Storm-Mathisen J, Leknes AK, Bore AT, Vaaland JL, Edminson P, Haug FMS, Ottersen OP (1983) First visualization of glutamate and GABA in neurones by immunocytochemistry. Nature 301:517–520

    Google Scholar 

  • Vollenweider FX, Cuénod M, Do KQ (1990) Effect of climbing fibre deprivation on release of endogenous aspartate, glutamate, and homocysteate in slices of rat cerebellar hemispheres and vermis. J Neurochem 54:1533–1540

    Google Scholar 

  • Wang B-L, Larsson L-I (1985) Simultaneous demonstration of multiple antigens by indirect immunofluorescence or immunogold staining: novel light and electron microscopial double and triple staining method employing primary antibodies from the same species. Histochemistry 83:47–56

    Google Scholar 

  • Wersäll J (1956) Studies on the structure and innervation of the sensory epithelium of the cristae ampullares in the guinea pig. Acta Otolaryngol Suppl (Stockh) 126:185

    Google Scholar 

  • Zhang N, Walberg F, Laake JH, Meldrum BS, Ottersen OP (1990) Aspartate-like and glutamate-like immunoreactivities in the inferior olive and climbing fibre system: a light microscopic and semiquantitative electron microscopic study in rat and baboon (Papio anubis). Neuroscience 38:61–80

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Usami, Si., Osen, K.K., Zhang, N. et al. Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate. Exp Brain Res 91, 1–11 (1992). https://doi.org/10.1007/BF00230008

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00230008

Key words

Navigation