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Gap junctional coupling and connexin immunoreactivity in rabbit retinal glia

Published online by Cambridge University Press:  09 March 2006

KATHLEEN R. ZAHS  and
PAUL W. CEELEN
KATHLEEN R. ZAHS
Affiliation:
Department of Physiology, University of Minnesota Medical School, Minneapolis, Minnesota
PAUL W. CEELEN
Affiliation:
Department of Neuroscience, University of Minnesota Medical School, Minneapolis, Minnesota
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Abstract

Gap junctions provide a pathway for the direct intercellular exchange of ions and small signaling molecules. Gap junctional coupling between retinal astrocytes and between astrocytes and Müller cells, the principal glia of vertebrate retinas, has been previously demonstrated by the intercellular transfer of gap-junction permeant tracers. However, functional gap junctions have yet to be demonstrated between mammalian Müller cells. In the present study, when the gap-junction permeant tracers Neurobiotin and Lucifer yellow were injected into a Müller cell via a patch pipette, the tracers transferred to at least one additional cell in more than half of the cases examined. Simultaneous whole-cell recordings from pairs of Müller cells in the isolated rabbit retina revealed electrical coupling between closely neighboring cells, confirming the presence of functional gap junctions between rabbit Müller cells. The limited degree of this coupling suggests that Müller cell–Müller cell gap junctions may coordinate the functions of small ensembles of these glial cells. Immunohistochemistry and immunoblotting were used to identify the connexins in rabbit retinal glia. Connexin30 (Cx30) and connexin43 (Cx43) immunoreactivities were associated with astrocytes in the medullary ray region of the retinas of both pigmented and albino rabbits. Connexin43 was also found in Müller cells, but antibody recognition differed between astrocytic and Müller cell connexin43.

Keywords

Müller cellAstrocyteElectrical couplingDye coupling
Type
Research Article
Information
Visual Neuroscience , Volume 23 , Issue 1 , January 2006 , pp. 1 - 10
Copyright
2006 Cambridge University Press

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References

REFERENCES

Altevogt, B.M., Kleopa, K.A., Postma, F.R., Scherer, S.S., & Paul, D.L. (2002). Connexin29 is uniquely distributed within myelinating glial cells of the central and peripheral nervous systems. Journal of Neuroscience 22, 6458–6470.Google Scholar
Ball, A.K. & McReynolds, J.S. (1998). Localization of gap junctions and tracer coupling in retinal Müller cells. Journal of Comparative Neurology 393, 48–57.3.0.CO;2-Q>CrossRefGoogle Scholar
Becker, D., Bonness, V., & Mobbs, P. (1998). Cell coupling in the retina, patterns and purpose. Cell Biology International 22, 781–792.CrossRefGoogle Scholar
Bennett, M.V.L. & Zukin, R.S. (2004). Electrical coupling and neuronal synchronization in the mammalian brain. Neuron 41, 495–511.CrossRefGoogle Scholar
Bevans, C.G., Kordel, M., Rhee, S.K., & Harris, A.L. (1998). Isoform composition of connexin channels determines selectivity among second messengers and uncharged molecules. Journal of Biological Chemistry 273, 2808–2816.CrossRefGoogle Scholar
Burris, C., Klug, K., Ngo, I.T., Sterling, P., & Schein, S. (2002). How Müller glial cells in macaque fovea coat and isolate the synaptic terminals of cone photoreceptors. Journal of Comparative Neurology 453, 100–111.CrossRefGoogle Scholar
Ceelen, P.W., Lockridge, A., & Newman, E.A. (2001). Electrical coupling between glial cells in the rat retina. Glia 35, 1–13.CrossRefGoogle Scholar
Charles, A.C., Dirksen, E.R., Merrill, J.E., & Sanderson, M.J. (1993). Mechanisms of intercellular calcium signaling in glial cells studied with dantrolene and thapsigargin. Glia 7, 134–145.CrossRefGoogle Scholar
Dermietzel, R. & Spray, D.C. (1993). Gap junctions in the brain: Where, what type, how many and why? Trends in Neuroscience 16, 186–192.Google Scholar
Dermietzel, R., Traub, O., Hwang, T.K., Beyer, E., Bennett, M.V.L., Spray, D.C., & Willecke, K. (1989). Differential expression of three gap junction proteins in developing and mature brain tissues. Proceedings of the National Academy of Sciences of the U.S.A. 86, 10148–10152.CrossRefGoogle Scholar
Dermietzel, R., Farooq, M., Kessler, J.A., Althaus, H., Hertzberg, E.L., & Spray, D.C. (1997). Oligodendrocytes express gap junction proteins connexin32 and connexin45. Glia 20, 101–114.3.0.CO;2-C>CrossRefGoogle Scholar
Doble, B.W., Chen, Y., Bosc, D.G., Litchfield, D.W., & Kardami, E. (1996). Fibroblast growth factor-2 decreases metabolic coupling and stimulates phosphorylation as well as masking of connexin43 epitopes in cardiac myocytes. Circulation Research 79, 647–658.CrossRefGoogle Scholar
Eugenin, E.A., Eckardt, D., Theis, M., Willecke, K., Bennett, M.V., & Saez, J.C. (2001). Microglia at brain stab wounds express connexin 43 and in vitro form functional gap junctions after treatment with interferon-γ and tumor necrosis factor-α. Proceedings of the National Academy of Sciences of the U.S.A. 98, 4190–4195.CrossRefGoogle Scholar
Giaume, C., Tabernero, A., & Medina, J.M. (1997). Metabolic trafficking through astrocytic gap junctions. Glia 21, 114–123.3.0.CO;2-V>CrossRefGoogle Scholar
Goldberg, G.S., Lampe, P.D., & Nicholson, B.J. (1999). Selective transfer of endogenous metabolites through gap junctions composed of different connexins. Nature Cell Biology 1, 457–459.CrossRefGoogle Scholar
Higgs, M.H. & Lukasiewicz, P.D. (1999). Glutamate uptake limits synaptic excitation of retinal ganglion cells. Journal of Neuroscience 19, 3691–3700.Google Scholar
Holländer, H., Makarov, F., Dreher, Z., Driel, D.V., Chan-Ling, T., & Stone, J. (1991). Structure of the macroglia of the retina, sharing and division of labour between astrocytes and Müller cells. Journal of Comparative Neurology 313, 587–603.CrossRefGoogle Scholar
Hossain, M.Z., Murphy, L.J., & Nagy, J.I. (1994a). Phosphorylated forms of connexin43 predominate in rat brain: Demonstration by rapid inactivation of brain metabolism. Journal of Neurochemistry 62, 2394–2403.Google Scholar
Hossain, M.Z., Sawchuk, M.A., Murphy, L.J., Hertzberg, E.L., & Nagy, J.I. (1994b). Kainic acid induced alterations in antibody recognition of connexin43 and loss of astrocytic gap junctions in rat brain. Glia 10, 250–265.Google Scholar
Johansson, K., Bruun, A., & Ehinger, B. (1999). Gap junction protein connexin43 is heterogeneously expressed among glial cells in the adult rabbit retina. Journal of Comparative Neurology 407, 395–403.3.0.CO;2-3>CrossRefGoogle Scholar
Jones, P.S., Elias, J.M., & Schecter, N. (1986). An improved method for embedding retina for cryosectioning. Journal of Histotechnology 9, 181–182.CrossRefGoogle Scholar
Kanno, S. & Saffitz, J.E. (2001). The role of myocardial gap junctions in electrical conduction and arrhythmogenesis. Cardiovascular Pathology 10, 169–177.CrossRefGoogle Scholar
Kettenmann, H. & Ransom, B.R. (1988). Electrical coupling between astrocytes and between oligodendrocytes studied in mammalian cell cultures. Glia 1, 64–73.CrossRefGoogle Scholar
Kunzelmann, P., Blumcke, I., Traub, O., Dermietzel, R., & Willecke, K. (1997). Coexpression of connexin45 and -32 in oligodendrocytes of rat brain. Journal of Neurocytology 26, 17–22.CrossRefGoogle Scholar
Kunzelmann, P., Schroder, W., Traub, O., Steinhauser, C., Dermietzel, R., & Willecke, K. (1999). Late onset and increasing expression of the gap junction protein connexin30 in adult murine brain and long-term cultured astrocytes. Glia 25, 111–119.3.0.CO;2-I>CrossRefGoogle Scholar
Laird, D.W., Puranam, K.L., & Revel, J.P. (1991). Turnover and phosphorylation dynamics of connexin43 gap junction protein in cultured cardiac myocytes. Biochemical Journal 273, 67–72.CrossRefGoogle Scholar
Laskawi, R., Rohlmann, A., Landgrebe, M., & Wolff, J. (1997). Rapid astroglial reactions in the motor cortex of adult rats following peripheral facial nerve lesions. European Archives of Otorhinolaryngology 254, 81–85.CrossRefGoogle Scholar
Li, W.E.I. & Nagy, J.I. (2000). Activation of fibres in rat sciatic nerve alters phosphorylation state of connexin-43 at astrocytic gap junctions in spinal cord: Evidence for junction regulation by neuronal-glial interactions. Neuroscience 97, 113–123.CrossRefGoogle Scholar
Li, W.E.I., Ochalski, P.A.Y., Hertzberg, E.L., & Nagy, J.I. (1998). Immunorecognition, ultrastructure and phosphorylation status of astrocytic gap junctions and connexin43 in rat brain after cerebral focal ischaemia. European Journal of Neuroscience 10, 2444–2463.CrossRefGoogle Scholar
Linser, P.J., Sorrentino, M., & Moscona, A.A. (1984). Cellular compartmentalization of carbonic anhydrase-C and glutamine synthetase in developing and mature mouse neural retina. Brain Research 315, 65–71.CrossRefGoogle Scholar
Marrero, H. & Orkand, R.K. (1996). Nerve impulses increase glial intercellular permeability. Glia 16, 285–289.3.0.CO;2-W>CrossRefGoogle Scholar
Matesic, D., Tillen, T., & Sitaramayya, A. (2003). Connexin 40 expression in bovine and rat retinas. Cell Biology International 27, 89–99.CrossRefGoogle Scholar
Menichella, D.M., Goodenough, D.A., Sirkowski, E., Scherer, S.S., & Paul, D.L. (2003). Connexins are critical for normal myelination in the CNS. Journal of Neuroscience 23, 5963–5973.Google Scholar
Mobbs, P., Brew, H., & Attwell, D. (1988). A quantitative analysis of glial cell coupling in the retina of the axolotl (Ambystoma mexicanum). Brain Research 460, 235–245.CrossRefGoogle Scholar
Müller, T., Moller, T., Neuhaus, J., & Kettenmann, H. (1996). Electrical coupling among Bergmann glial cells and its modulation by glutamate receptor activation. Glia 17, 274–284.3.0.CO;2-#>CrossRefGoogle Scholar
Musil, L.S. & Goodenough, D.A. (1991). Biochemical analysis of connexin43 intracellular transport, phosphorylation, and assembly into gap junctional plaques. Journal of Cell Biology 115, 1357–1374.CrossRefGoogle Scholar
Nadarajah, B., Thomaidou, D., Evans, W.H., & Parnavelas, J.G. (1996). Gap junctions in the adult cerebral cortex, regional differences in their distribution and cellular expression of connexins. Journal of Comparative Neurology 376, 326–342.3.0.CO;2-J>CrossRefGoogle Scholar
Nagy, J.I & Li, W.E.I. (2000). A brain slice model for in vitro analyses of astrocytic gap junction and connexin43 regulation: Actions of ischemia, glutamate and elevated potassium. European Journal of Neuroscience 12, 4567–4572.Google Scholar
Nagy, J.I., Patel, D., Ochalski, P.A., & Stelmack, G.L. (1999). Connexin30 in rodent, cat, and human brain: Selective expression in gray matter astrocytes, co-localization with connexin43 at gap junctions and late developmental appearance. Neuroscience 88, 447–468.CrossRefGoogle Scholar
Newman, E.A. (1985). Regulation of potassium levels by glial cells in the retina. Trends in Neuroscience 8, 156–159.CrossRefGoogle Scholar
Newman, E.A. (2001). Propagation of intercellular calcium waves in retinal astrocytes and Müller cells. Journal of Neuroscience 21, 2215–2223.Google Scholar
Newman, E.A. (2003). Glial cell inhibition of neurons by release of ATP. Journal of Neuroscience 23, 1659–1666.Google Scholar
Newman, E.A., Frambach, D.A., & Odette, L.L. (1984). Control of extracellular potassium levels by retinal glial cell potassium siphoning. Science 225, 1174–1175.CrossRefGoogle Scholar
Newman, E.A. & Zahs, K.R. (1997). Calcium waves in retinal glial cells. Science 275, 844–847.CrossRefGoogle Scholar
Newman, E.A. & Zahs, K.R. (1998). Modulation of neuronal activity by glial cells in the retina. Journal of Neuroscience 18, 4022–4028.Google Scholar
Niessen, H., Harz, H., Bender, P., Kramer, K., & Willecke, K. (2000). Selective permeability of different connexin channels to the second messenger inositol 1,4,5-trisphosphate. Journal of Cell Science 113, 1365–1372.Google Scholar
Odermatt, B., Wellershaus, K., Wallraff, A., Seifert, G., Degen, J., Euwens, C., Fuss, B., Bussow, H., Schilling, K., Steinhauser, C., & Willecke, K. (2003). Connexin 47 (Cx47)-deficient mice with enhanced green fluorescent protein reporter gene reveal predominant oligodendrocytic expression of Cx47 and display vacuolized myelin in the CNS. Journal of Neuroscience 23, 4549–4559.Google Scholar
Pow, D.V. (2001). Amino acids and their transporters in the retina. Neurochemistry International 38, 463–484.CrossRefGoogle Scholar
Rash, J.E., Yasumura, T., Davidson, K.G., Furman, C.S., Dudek, F.E., & Nagy, J.I. (2001). Identification of cells expressing Cx43, Cx30, Cx26, Cx32 and Cx36 in gap junctions of rat brain and spinal cord. Cell Communication and Adhesion 8, 315–320.CrossRefGoogle Scholar
Robinson, S.R., Hampson, E.C.G.M., Munro, M.N., & Vaney, D.I. (1993). Unidirectional coupling of gap junctions between neuroglia. Science 262, 1072–1074.CrossRefGoogle Scholar
Rohlmann, A., Laskawi, R., Hofer, A., Dermietzel, R., & Wolff, J.R. (1994). Astrocytes as rapid sensors of peripheral axotomy in the facial nucleus of rats. Neuroreport 5, 409–412.CrossRefGoogle Scholar
Rohlmann, A., Laskawi, R., Hofer, A., Dobo, E., Dermietzel, R., & Wolff, J.R. (1993). Facial nerve lesions lead to increased immunostaining of the astrocytic gap junction protein (connexin 43) in the corresponding facial nucleus of rats. Neuroscience Letters 154, 206–208.CrossRefGoogle Scholar
Rouach, N., Glowinski, J., & Giaume, C. (2000). Activity dependent neuronal control of gap-junctional communication in astrocytes. Journal of Cell Biology 149, 1513–1526.CrossRefGoogle Scholar
Saari, J.C., Huang, J.M., Possin, D.E., Fariss, R.N., Leonard, J., Garwin, G.G., Crabb, J.W., & Milam, A.H. (1997). Cellular retinaldehyde-binding protein is expressed by oligodendrocytes in optic nerve and brain. Glia 21, 259–268.3.0.CO;2-0>CrossRefGoogle Scholar
Schutte, M., Chen, S., Buku, A., & Wolosin, J. (1998). Connexin50, a gap junction protein of macroglia in the mammalian retina and visual pathway. Experimental Eye Research 66, 605–613.CrossRefGoogle Scholar
Simburger, E., Stang, A., Kremer, M., & Dermietzel, R. (1997). Expression of connexin43 mRNA in adult rodent brain. Histochemistry and Cell Biology 107, 127–137.CrossRefGoogle Scholar
Singh, D. & Lampe, P.D. (2003). Identification of connexin-43 interacting proteins. Cell Communication and Adhesion 10, 215–220.CrossRefGoogle Scholar
Solan, J.L. & Lampe, P.D. (2005). Connexin phosphorylation as a regulatory event linked to gap junction channel assembly. Biochimica et Biophysica Acta 1711, 154–163.CrossRefGoogle Scholar
Spray, D.C., Ginzberg, R.D., Morales, E.A., Gatmaitan, Z., & Arias, I.M.. (1986). Electrophysiological properties of gap junctions between dissociated pairs of rat hepatocytes. Journal of Cell Biology 103, 135–144.CrossRefGoogle Scholar
Stone, J., Makarov, F., & Holländer, H. (1995). The glial ensheathment of the soma and axon hillock of retinal ganglion cells. Visual Neuroscience 12, 273–279.CrossRefGoogle Scholar
Stout, C.E., Costantin, J.L., Naus, C.C., & Charles, A.C. (2002). Intercellular Ca2+ signaling in astrocytes via ATP release through connexin hemichannels. Journal of Biological Chemistry 277, 10482–10488.CrossRefGoogle Scholar
Trexler, E.B., Li, W., Mills, S.L., & Massey, S.C. (2001). Coupling from AII amacrine cells to ON cone bipolar cells is bidirectional. Journal of Comparative Neurology 437, 408–422.CrossRefGoogle Scholar
Yamamoto, T., Ochalski, A., Hertzberg, E.L., & Nagy, J.I. (1990). On the organization of astrocytic gap junctions in rat brain as suggested by LM and EM immunohistochemistry of connexin43 expression. Journal of Comparative Neurology 302, 853–883.CrossRefGoogle Scholar
Zahs, K.R., Kofuji, P., Meier, C., & Dermietzel, R. (2003). Connexin immunoreactivity in glial cells of the rat retina. Journal of Comparative Neurology 455, 531–546.CrossRefGoogle Scholar
Zahs, K.R. & Newman, E.A. (1997). Asymmetric gap junctional coupling between glial cells in the rat retina. Glia 20, 10–22.3.0.CO;2-9>CrossRefGoogle Scholar