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Localization of GABA- and GAD-like immunoreactivity in the turtle retina

Published online by Cambridge University Press:  02 June 2009

Lawrence B. Hurd II  and
William D. Eldred
Lawrence B. Hurd II
Affiliation:
Department of Biology, Boston University, Boston
William D. Eldred
Affiliation:
Department of Biology, Boston University, Boston
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Abstract

γ-aminobutyric acid (GABA) has been reported to be an important neurotransmitter in the retinas of many species. This immunocytochemical study detailed the localization of antigens resembling GABA and glutamic acid decarboxylase (GAD, an enzyme involved in the synthesis of GABA), in retinal neurons in the turtle, Pseudemys scripta elegans. GABA-like immunoreactivity was present within somata in the inner and outer regions of the inner nuclear layer, within somata in the ganglion cell layer, and in processes in the outer plexiform layer, inner plexiform layer, and ganglion cell axon layer. GAD-like immunoreactivity was found in somata in the inner and outer regions of the inner nuclear layer and in processes in the inner and outer plexiform layers. Cell counts indicated more somata with GABA-like than GAD-like immunoreactivity in the inner nuclear layer. Double-label studies showed that every somata in the inner nuclear layer which had GAD-like immunoreactivity also had GABA-like immunoreactivity, but that many somata had only GABA-like immunoreactivity.

The stratification of immunoreactivity within the inner plexiform layer was analyzed using a scanning densitometer. We described the strata within the inner plexiform layer such that S0 represented the inner nuclear layer/inner plexiform layer border and S100 represented the inner plexiform layer/ganglion cell layer border. Analysis of GAD-like labeling yielded seven distinct strata with peak densities at positions S8, S19, S28, S42, S59, S75, and S93. GABA-like labeling provided five distinct strata with peak densities at positions S17, S28, S67, S84, and S95. The strata with peaks of GABA-like immunoreactivity at S17 and S28 were in statistically identical locations to corresponding strata with GAD-like immunoreactivity. The strata with GABA-like immunoreactivity at S67, S84, and S95 did not have statistically identical peaks of correlated GAD-like immunoreactivity, although there were corresponding strata with GAD-like immunoreactivity nearby. Antiserum directed against GABA failed to produce labeled strata at positions corresponding to the strata with GAD-like immunoreactivity at S8 and S42.

In summary, our results indicated that the antisera we used, which were directed against GABA and GAD, produced significantly different labeling in the inner nuclear layer, inner plexiform layer, and the ganglion cell body and axon layers of the turtle retina. Until the physiological significance of these differences is resolved, studies employing these markers to investigate the function of GABA in the turtle retina should be interpreted with caution.

Keywords

NeurotransmitterImmunocytochemistryInner plexiform layerStratificationGanglion cell layer
Type
Research Articles
Information
Visual Neuroscience , Volume 3 , Issue 1 , July 1989 , pp. 9 - 20
Copyright
Copyright © Cambridge University Press 1989

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References

Agardh, E., Bruun, A., Ehinger, B. & Storm-Mathisen, J. (1986). GABA immunoreactivity in the retina. Investigative Ophthalmology and Visual Science 27, 674–678.Google ScholarPubMed
Agardh, E., Ehinger, B. & Wu, J.-Y. (1987 a). GABA- and GAD-like immunoreactivity in the primate retina. Histochemistry 86, 485–490.CrossRefGoogle ScholarPubMed
Agardh, E., Bruun, A., Ehinger, B., Ekstrom, P., van Veen, T. & Wu, J.-Y. (1987 b). Gamma-aminobutyric acid- and glutamic acid decarboxylase-immunoreactive neurons in the retina of different vertebrates. Journal of Comparative Neurology 258, 622–630.CrossRefGoogle ScholarPubMed
Ball, A.K. & Brandon, C. (1986). Localization of [3H]-GABA, -muscimol, and -glycine in goldfish retinas stained for glutamate decarboxylase. Journal of Neuroscience 6, 1621–1627.CrossRefGoogle ScholarPubMed
Bolz, J. & McGuire, B.A. (1985). GABA-like immunoreactivity in horizontal cells of the cat retina. Society for Neuroscience Abstracts 11, 1215.Google Scholar
Brandon, C. (1985). Retinal GABA neurons: localization in vertebrate species using an antiserum to rabbit brain glutamate decarboxylase. Drain Research 344, 286–295.Google ScholarPubMed
Brandon, C., Lam, D.M.K. & Wu, J.-Y. (1979). The γ-aminobutyric acid system in rabbit retina: localization by immunocytochemistry and autoradiography. Proceedings of the National Academy of Sciences of the U.S.A. 76, 3557–3561.CrossRefGoogle ScholarPubMed
Brandon, C., Lam, D.M.K., Su, Y.Y.T. & Wu, J.-Y. (1980). Immunocytochemical localization of GABA neurons in the rabbit and frog retina. Brain Research Bulletin 5, 21–29.CrossRefGoogle Scholar
Cajal, S.R.Y. (1933). Die Retina der Wirbeltiere. Wiesbaden: Bergmann. (Translation: Thorpe S.A. & Glickstein M. (1972). The Structure of the Retina. Springfield: Thomas).Google Scholar
Denner, L.A. & Wu, J.-Y. (1985). Two forms of rat brain glutamic acid decarboxylase differ in their dependence on free pyridoxal phosphate. Journal of Neurochemistry 44, 957–965.CrossRefGoogle ScholarPubMed
Eldred, W.D., Zucker, C., Karten, H.J. & Yazulla, S. (1983). Comparison of fixation and penetration-enhancement techniques for use in ultrastructural immunocytochemistry. Journal of Histochemistry and Cytochemistry 31, 285–292.CrossRefGoogle ScholarPubMed
Famiglietti, E.V. Jr. & Vaughn, J.E. (1981). Golgi-impregnated amacrine cells and GABAergic retinal neurons: a comparison of dendritic, immunocytochemical, and histochemical stratification in the inner plexiform layer of the rat retina. Journal of Comparative Neurology 197, 129–139.CrossRefGoogle ScholarPubMed
Hendrickson, A., Ryan, M., Nobel, B. & Wu, J.-Y. (1985). Colocalization of [3H]-muscimol and antisera to GABA and glutamic acid decarboxylase within the same neurons in monkey retina. Brain Research 348, 391–396.CrossRefGoogle ScholarPubMed
Hughes, A. (1985). New perspectives in retinal organization. In Progress in Retinal Research, Vol. 4, ed. Osborne, N.N. & Chader, G.J., pp. 243–313. Oxford: Pergamon Press.Google Scholar
Lam, D.M.K., Su, Y.Y.T., Swain, L., Marc, R.E., Brandon, C. & Wu, J.-Y. (1979). Immunocytochemical localization L-glutamic acid decarboxylase in the goldfish retina. Nature 278, 565–567.CrossRefGoogle ScholarPubMed
Lam, D.M.K., Su, Y.Y.T., Chin, C.A., Brandon, C., Wu, J.-Y., Marc, R.E. & Lasater, E.M. (1980). GABAergic horizontal cells in the teleost retina. Brain Research Bulletin 5, 137–140.CrossRefGoogle Scholar
Leeper, H.F. (1978). Horizontal cells of the turtle retina. I. Light microscopy of Golgi preparations. Journal of Comparative Neurology 182, 777–794.CrossRefGoogle ScholarPubMed
Lin, C.-T., Li, H.-Z. & Wu, J.-Y. (1983). Immunocytochemical localization of L-glutamate decarboxylase, gamma-aminobutyric acid transaminase, cysteine sulfinic acid decarboxylase, aspartate aminotransferase, and somatostatin in rat retina. Brain Research 270, 273–283.CrossRefGoogle ScholarPubMed
Lin, C.-T., Song, G.-X. & Wu, J.-Y. (1985). Ultrastructural demonstration of L-glutamate decarboxylase and cysteine sulfinic add decarboxylase in rat retina by immunocytochemistry. Brain Research 33, 71–80.Google Scholar
Marc, R.E. (1986). Neurochemical stratification in the inner plexiform layer of the vertebrate retina. Vision Research 26, 223–238.CrossRefGoogle ScholarPubMed
Morgan, W.W. (1985). GABA: a potential neurotransmitter in retina. In Retinal Transmitters and Modulators: Models for the Brain, Vol. 2, ed. Morgan, W.W., pp. 63–96. Boca Raton, Florida: CRC Press.Google Scholar
Mosinger, J.L. & Yazulla, S. (1985). Colocalization of GAD immunoreactivity and [3H]-GABA uptake in amacrine cells of rabbit retina. Journal of Comparative Neurology 240, 396–406.CrossRefGoogle ScholarPubMed
Mosinger, J.L. & Yazulla, S. (1987). Double-label analysis of GADand GABA-like immunoreactivity in the rabbit retina. Vision Research 27(1), 23–30.CrossRefGoogle Scholar
Mosinger, J.L., Studholme, K.M. & Yazulla, S. (1985). Immunocytochemical localization of GABA in the retina: a species comparison. Investigative Ophthalmology and Visual Science (Suppl.) 26, 94 (Abstract).Google Scholar
Mosinger, J.L., Yazulla, S. & Studholme, K.M. (1986). GABA-like immunoreactivity in the vertebrate retina: a species comparison. Experimental Eye Research 42, 631–644.CrossRefGoogle ScholarPubMed
Nakamura, Y., McGuire, B.A. & Sterling, P. (1980). Interplexiform cell in cat retina: identification by uptake of γ-[3H]aminobutyric acid and serial reconstructions. Proceedings of the National Academy of Sciences of the U.S.A. 77, 658–661.CrossRefGoogle Scholar
Oertel, W.H., Tappaz, I.J., Kopin, I.J., Ranson, D.H. & Schmechel, D.E. (1980). Production of an antiserum to rat brain glutamate (GAD)/cysteine sulfinate (CSD) decarboxylase. Brain Research Bulletin 5, 713–719.CrossRefGoogle Scholar
Osborne, N.N., Patel, S., Beaton, D.W. & Neuhoff, V. (1986). GABA neurones in the retinas of different species and their postnatal development in situ and in culture in the rabbit retina. Cell and Tissue Research 243, 117–123.CrossRefGoogle ScholarPubMed
Pourcho, R.G., Goebel, D.J. & McReynolds, J.S. (1984). Autoradiographic studies of [3H]-glycine, [3H]-GABA, and [3H]-muscimol uptake in the mudpuppy retina. Experimental Eye Research 39, 69–81.CrossRefGoogle Scholar
Ryan, M. & Hendrickson, A. (1987). Interplexiform cells in the macaque monkey retina. Experimental Eye Research 45, 57–66.CrossRefGoogle ScholarPubMed
Spink, D.C., Wu, S.J. & Martin, D.L. (1983). Multiple forms of glutamate decarboxylase in porcine brain. Journal of Neurochemistry 40, 1113–1119.CrossRefGoogle ScholarPubMed
Storm-Mathisen, J., Leknes, A.K., Bore, A.T., Vaaland, J.L., Edminson, P., Haug, F.M.S. & Ottersen, O.P. (1983). First visualization of glutamate and GABA in neurones by immunocytochemistry. Nature 301, 517–520.CrossRefGoogle ScholarPubMed
Studholme, K.M., Yazulla, S. & Wu, J.-Y. (1985). A double-label analysis of GABAergic amacrine cells in the goldfish retina. Investigative Ophthalmology and Visual Science (Suppl.) 26, 95 (Abstract).Google Scholar
Vaney, D.I. & Young, H.M. (1988). GABA-like immunoreactivity in cholinergic amacrine cells of the rabbit retina. Brain Research 438, 369–373.CrossRefGoogle ScholarPubMed
Vaughn, J.E., Barber, R.P., Saito, K., Roberts, E. & Famiglietti, E.V. Jr. (1978). Immunocytochemical identification of GABAergic neurons in rat retina. Anatomical Record 190, 571–572 (Abstract).Google Scholar
Vaughn, J.E., Famiglietti, E.V. Jr., Barber, R.P., Saito, K., Roberts, E. & Ribak, C.E. (1981). GABAergic amacrine cells in the rat retina: immunocytochemical identification and synaptic connectivity. Journal of Comparative Neurology 197, 113–127.CrossRefGoogle ScholarPubMed
Voaden, M.J., Marshall, J., & Murani, N. (1974). The uptake of 3H-γ-aminobutyric acid and 3H-glycine by the isolated retina of the frog. Brain Research 67, 115–132.CrossRefGoogle Scholar
Voaden, M.J. (1976). Gamma-aminobutyric acid and glycine as retinal neurotransmitters. In Transmitters in the Visual Process, ed. Bonting, S.E., pp. 107–125. Oxford: Pergamon Press.CrossRefGoogle Scholar
Wenthold, R.E., Zemple, I.M., Parakkal, M.H., Reeks, K.A. & Altschuler, R.A. (1986). Immunocytochemical localization of GABA in the cochlear nucleus of the guinea pig. Brain Research 380, 7–18.CrossRefGoogle ScholarPubMed
Wu, J.-Y., Brandon, C., Su, Y.Y. & Lam, D.M.K. (1981). Immunocytochemical and autoradiographic localization of GABA system in the vertebrate retina. Molecular and Cell Biochemistry 39, 229–238.CrossRefGoogle ScholarPubMed
Yazulla, S. (1986). GABAergic mechanisms in the retina. In Progress in Retina Research, Vol. 5, ed. Osborne, N.N. & Chader, G.J., pp. 1–52. Oxford: Pergamon Press.Google Scholar
Yazulla, S., Studholme, K. & Wu, J.-Y. (1986). Comparative distribution of [3H]-GABA uptake and GAD immunoreactivity in goldfish retinal amacrine cells: a double-label analysis. Journal of Comparative Neurology 244, 149–162.CrossRefGoogle ScholarPubMed
Yu, B.C.-Y., Watt, C.B., Lam, D.M.K. & Fry, K.R. (1988). GABAergic ganglion cells in the rabbit retina. Brain Reseatch 439, 376–382.CrossRefGoogle ScholarPubMed
Zucker, C., Yazulla, S. & Wu, J.-Y. (1984). Non-correspondence of [3H]-GABA uptake and GAD localization in goldfish amacrine cells. Brain Research 298, 154–158.CrossRefGoogle ScholarPubMed