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Localization of vanilloid receptor 1 (TRPV1/VR1)-like immunoreactivity in goldfish and zebrafish retinas: Restriction to photoreceptor synaptic ribbons

  • Published:
Journal of Neurocytology

Abstract

The vanilloid receptor type 1 (TRPV1/VR1) is a non-specific calcium-permeable ionotropic cation channel expressed in the peripheral sensory system as well as in the central nervous system. An endogenous ligand for TRPV1 is arachidonoyl ethanolamide (anandamide), which also activates the metabotropic cannabinoid receptor 1 (CB1). Previous studies in this laboratory reported CB1 receptors and CB1-mediated effects on voltage-gated currents in goldfish cones and bipolar cells. In this study, we show TRPV1-like-immunoreactivity (TRPV1-L-IR) by immunoblot analysis of goldfish retina and rat brain homogenates with a guinea pig polyclonal antibody against the C-terminus of the rat TRPV1. Light-level immunocytochemistry showed restriction of the guinea pig-TRPV1 antibody to a very narrow band in the outer plexiform layer in goldfish and zebrafish retinas. However, no immunoreactivity was detected using rabbit-polyclonal antibodies against the C or N-termini of the rat TRPV1. Pre and post-embedding electron microscopy (EM) immunocytochemistry revealed that TRPV1-L-IR (using the guinea pig antibody) was restricted to synaptic ribbons of all cones and many rods, but never was observed at the synaptic ribbons of bipolar cells. While pre-embedded tissue showed diffuse label associated only with photoreceptor-synaptic ribbons, analysis of post-embedded tissue showed label tightly restricted to synaptic ribbons of all cones and many rods. Oblique sections showed that immunogold particles were confined to the outer electron dense region of the ribbons, with few or no gold particles in the ribbon core or associated with tethers or vesicles. Although TRPV1-L-IR described here, does not necessarily represent TRPV1 antigen associated with synaptic ribbons, these data provide an unequivocal label with which to study the functional dynamics of ribbon formation and degradation in teleost photoreceptors.

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References

  • Ahluwalia, J., Urban, L., Bevan, S. & Nagy, I. (2003) Anandamide regulates neuropeptide release from capsaicin-sensitive primary sensory neurons by activating both the cannabinoid 1 receptor and the vanilloid receptor 1 in vitro. European Journal of Neuroscience 17, 2611–2618.

    Google Scholar 

  • Ahluwalia, J., Urban, L., Capogna, M., Bevan, S. & Nagy, I. (2000) Cannabinoid 1 receptors are ex-pressed in nociceptive primary sensory neurons. Neuro-science 100, 685–688.

    Google Scholar 

  • Allwardt, B. A., Lall, A. B., Brockerhoff, S. E. & Dowling, J. E. (2001) Synapse formation is arrested in retinal photoreceptors of the zebrafish nrc mutant. Journal of Neuroscience 21, 2330–2342.

    Google Scholar 

  • Balkema, G. W. & Nguyen, T. H. (1999) Co-localization of clathrin adapter proteins with photore-ceptor synaptic ribbons in the OPL of the mouse. Society for Neuroscience Abstracts 25, 135.

    Google Scholar 

  • Balkema, G. W. & Rizkalla, R. (1996) Ultrastructural localization of a synaptic ribbon protein recognized by antibody B16. Journal of Neurocytology 25, 565–571.

    Google Scholar 

  • BrandstÄtter, J. H., Fletcher, E. L., Garner, C. C., Gundelfinger, E. D. & wÄssle, H. (1999) Differential expression of the presynaptic cy-tomatrix protein bassoon among ribbon synapses in the mammalian retina. European Journal of Neuroscience 11, 3683–3693.

    Google Scholar 

  • Caterina, M. J., Schumacher, M., Tominaga, M., Rosen, T. A., Levine, J. D. & Julius, D. (1997) The capsaicin receptor: A heat-activated ion channel in the pain pathway. Nature 389, 819–824.

    Google Scholar 

  • Couet, J., Belanger, M. M., Roussel, E. & Drolet, M. C. (2001) Cell biology of caveolae and caveolin. Advanced Drug Delivery Reviews 49, 223–235.

    Google Scholar 

  • De Juan, J., García, M. & Cuenca, N. (1996) For-mation and dissolution of spinules and changes in ne-matosome size require optic nerve integrity in black bass (Micropterus salmoidess)retina. Brain Research 707, 213–220.

    Google Scholar 

  • De Petrocellis, L., Bisogno, T., Maccarrone, M., Davis, J. B., Finazzi-Agro, A. & Dimarzo, V. (2001) The activity of anandamide at vanilloid VR1 re-ceptors requires facilitated transport across the cell mem-brane and is limited by intracellular metabolism. Journal of Biological Chemistry 276, 12856–12863.

    Google Scholar 

  • Dick, O., Hack, I., Altrock, W. D., Garner, C. C., Gundelfinger, E. D. & BrandstÄtter, J. H. (2001) Localization of the presynaptic cytomatrix protein piccolo at ribbon and conventional synapses in the rat retina: Comparison with bassoon. Journal of Comparative Neurology 439, 224–234.

    Google Scholar 

  • Dimarzo, V., Depetrocellis, L., Fezza, F., Ligresti, A. & Bisogno, T. (2002) Anandamide receptors. Prostaglandins Leukotrienes and Essential Fatty Acids 66, 377–391.

    Google Scholar 

  • Fan, S. F. & Yazulla, S. (2003) Biphasic modulation of voltage-dependent currents of retinal cones by cannabinoid CB1 agonist, WIN 55212–2. Visual Neuroscience 20, 177–188.

    Google Scholar 

  • Glaser, S. T., Studholme, K. M., Fatade, R., Yazulla, S., Abumrad, N. A. & Deutsch, D. G. (2002) Is there an anandamide transporter. Burlington, VT, International Cannabinoid Research Society. 2002 Symposium on the Cannabinoids. Page 9. http://cannabinoidsociety.org/HTML/ ICRS.2002.symposium.pdf

  • GrÜn, G. (1980) Developmental dynamic in synaptic ribbons of retinal receptor cells (Tilapia, Xenopus). Cell and Tissue Research 207, 331–339.

    Google Scholar 

  • Gumbleton, M., Abulrob, A. G. & Campbell, L. (2000) Caveolae: An alternative membrane transport compartment. Pharmaceutical Research 17, 1035–1048.

    Google Scholar 

  • Gunthorpe, M. J., Benham, C. D., Randall, A. & Davis, J. B. (2002) The diversity in the vanilloid (TRPV).Synaptic ribbons contain TRPV1-like-IR 451 receptor family of ion channels. Trends in Pharmacological Science 23, 183–191.

    Google Scholar 

  • Guo, A., Vulchanova, L., Wang, J., Li, X. & Elde, R. (1999) Immunocytochemical localization of the vanilloid receptor 1 (VR1): Relationship to neuropeptides, the P2X3 purinoceptor and IB4 binding sites. European Journal of Neuroscience 11, 946–958.

    Google Scholar 

  • Kachi, S., Yamazaki, A. & Usukura, J. (2001) Localization of caveolin-1 in photoreceptor synaptic ribbons. Investigative Ophthalmology & Visual Science 42, 850–852.

    Google Scholar 

  • Kedei, N., Szabo, T., Lile, J. D., Treanor, J. J., Olah, Z., Iadarola, M. J. & Blumberg, P. M. (2001) Analysis of the native quaternary structure of vanilloid receptor 1. Journal of Biological Chemistry 276, 28613–28619.

    Google Scholar 

  • Klooster, J., Studholme, K. M. & Yazulla, S. (2001) Localization of the AMPA subunit GluR2 in the outer plexiform layer of goldfish retina. Journal of Com-parative Neurology 441, 155–167.

    Google Scholar 

  • Ladman, A. J. (1958) The fine structure of the rod-bipolar cell synapse in the retina of the albino rat. Journal of Bio-physical and Biochemical Cytology 4, 459–466.

    Google Scholar 

  • Larison, R. D. & Bremiller, R. (1990) Early onset of phenotype and cell patterning in the embryonic zebrafish retina. Development 109, 567–576.

    Google Scholar 

  • Liu, M., Liu, M. C., Magoulas, C., Priestley, J. V. & Willmott, N. J. (2003) Versatile regulation of cytosolic Ca 2+ by vanilloid receptor I in rat dorsal root gan-glion neurons. Journal of Cell Biology 278, 5462–5472.

    Google Scholar 

  • Magga, J. M., Kay, J. G., Davy, A., Poulton, N. P., Robbins, S. M. & Braun, J. E. A. (2002) ATP dependence of the SNARE/caveolin 1 interaction in the hippocampus. Biochemical and Biophysical Research Communications 291, 1232–1238.

    Google Scholar 

  • McAllister, S. D. & Glass, M. (2002) CB1 and CB2 receptor-mediated signalling: A focus on endocannabinoids. Prostaglandins Leukotrienes and Essential Fatty Acids 66, 161–171.

    Google Scholar 

  • Mezey, E., Toth, A., Cortright, D. N., Arzubi, M. K., Krause, J. E., Elde, R., Guo, A., Blumberg, P. M. & Szallasi, A. (2000) Distribution of mRNA for vanilloid receptor subtype 1 (VR1), and VR1-like immunoreactivity, in the central nervous system of the rat and human. Proceedings of the National Academy of Science of the U.S.A. 97, 3655–3660.

    Google Scholar 

  • Minke, B. & Cook, B. (2002) TRP channel proteins and signal transduction. Physiological Reviews 82, 429–472.

    Google Scholar 

  • Muresan, V., Lyass, A. & Schnapp, B. J. (1999) The kinesin motor KIF3A is a component of the presynaptic ribbon in vertebrate photoreceptors. Journal of Neuro-science 19, 1027–1037.

    Google Scholar 

  • Nguyen, T. H. & Balkema, G. W. (1999) Antigenic epitopes of the photoreceptor synaptic ribbon. Journal of Comparative Neurology 413, 209–218.

    Google Scholar 

  • Premkumar, L. S. & Ahern, G. P. (2000) Induction of vanilloid channel activity by protein kinase C. Nature 408, 985–990.

    Google Scholar 

  • Rao-Mirotznik, R., Harkins, A. B., Buchsbaum, G. & Sterling, P. (1995) Mammalian rod terminal: Architecture of a binary synapse. Neuron 14, 561–569.

    Google Scholar 

  • Roof, D., Hayes, A., Hardenbergh, G. & Adamian, M. (1991) A 52 kD cytoskeletal protein from retinal rod photoreceptors is related to erythrocyte dematin. Investigative Ophthalmology & Visual Science 32, 582–593.

    Google Scholar 

  • Roof, D., Hayes, A., Adamian, M., Chishti, A.H. & Li, T. (1997) Molecular characterization of abLIM, a novel actin-binding and double zinc finger protein. Journal of Cell Biology 138, 575–588.

    Google Scholar 

  • Rosenbaum, T., Awaya, M. & Gordon, S. E. (2002) Subunit modification and association in VR1 ion Channels. BioMed Central Neuroscience 3.

  • Schmitz, F., Kirsch, M. & Wagner, H.-J. (1989) Calcium modulated synaptic ribbon dynamics in cone photoreceptors: A pharmacological and electron spectroscopic study. European Journal of Cell Biology 49, 207–212.

    Google Scholar 

  • Schmitz, F., Konigstorfer, A. & SÜdhof, T. C. (2000) RIBEYE, a component of synaptic ribbons: A protein's journey through evolution provides insight into synaptic ribbon function. Neuron 28, 857–872.

    Google Scholar 

  • Sjostrand, F. S. (1958) Ultrastructure of retinal rod synapses of the guinea pig eye as revealed by three-dimensional reconstructions from serial synapses. Journal of Ultrastructure Research 2, 122–170.

    Google Scholar 

  • Smart, D., Gunthorpe, M. J., Jerman, J. C., Nasir, S., Gray, J., Muir, A. I., Chambers, J. K., Randall, A. D. & Davis, J. B. (2000) The endogenous lipid anandamide is a full agonist at the human vanilloid receptor (hVR1). British Journal of Pharmacology 129, 227–230.

    Google Scholar 

  • Smart, D., Jerman, J. C., Gunthorpe, M. J., Brough, S. J., Ranson, J., Cairns, W., Hayes, P. D., Randall, A. D. & Davis, J. B. (2001) Characterisation using FLIPR of human VR1 receptor pharmacology. European Journal of Pharmacology 417, 51–58.

    Google Scholar 

  • Stell, W. K. (1976) Functional polarization of horizontal cell dendrites in goldfish retina. Investigative Ophthalmology 15, 895–908.

    Google Scholar 

  • Sternberger, L. A. (1979) Immunocytochemistry, 2 edition. New York: John Wiley & Sons.

    Google Scholar 

  • Suzuki, S. & Kaneko, A. (1990), Identification of bipolar cell subtypes by protein kinase C-like immunoreactivity in the goldfish retina. Visual Neuroscience 5, 223–230.

    Google Scholar 

  • Szallasi, A. & Dimarzo, V. (2000) New perspectives on enigmatic vanilloid receptors. Trends in Neuroscience 23, 491–497.

    Google Scholar 

  • Tominaga, M., Caterina, M. J., Malmberg, A. B., Rosen, T. A., Gilbert, H., Skinner, K., Raumann, B. E., Basbaum, A. I. & Julius, D. (1998) The cloned capsaicin receptor integrates multiple pain-producing stimuli. Neuron 21, 531–543.

    Google Scholar 

  • Van Den Pol, A. N. & GÖrcs, T. (1988) Glycine and glycine receptor immunoreactivity in brain and spinal cord. Journal of Neuroscience 8, 472–492.

    Google Scholar 

  • Vollrath, L., Meyer, A. & Buschmann, F. (1989) Ribbon synapses of the mammalian retina contain two types of synaptic bodies--ribbons and spheres. Journal of Neurocytology 18, 115–120.

    Google Scholar 

  • Vollrath, L. & Spiwoks-Becker, I. (1996) Plasticity of retinal ribbon synapses. Microscopy Research and Technique 35, 472–487.

    Google Scholar 

  • Wagner, H.-J. (1973). Darkness-induced reduction of the number of synaptic ribbons in fish retina. Nature New Biology 246, 53–55.

    Google Scholar 

  • Wagner, H.-J. & Ali, M. A. (1977) Cone synaptic ribbons and retinomotor changes in the brook trout, Salvelinus fontinalis (Salmonidae, Teleostei), under various experimental conditions. Canadian Journal of Zoology 55, 1684–1691.

    Google Scholar 

  • Wang, Y., Okamoto, M., Schmitz, F., Hofmann, K. & SÜdhof, T. C. (1997) Rim is a putative Rab3 effector in regulating synaptic-vesicle fusion. Nature 388, 593–598.

    Google Scholar 

  • Wisnoskey, B. J., Sinkins, W.G. & Schilling, W. P. (2003) Activation of vanilloid receptor type I in the endoplasmic reticulum fails to activate store-operated Ca2+ entry. Biochemical Journal 372, 517–528.

    Google Scholar 

  • Yao, P. J., Coleman, P. D. & Calkins, D. J. (2002) High-resolution localization of clathrin assembly protein AP180 in the presynaptic terminals of mammalian neurons. Journal of Comparative Neurology 447, 152–162.

    Google Scholar 

  • Yazulla, S. & Studholme, K. M. (2001) Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry. Journal of Neurocytology 30, 551–592.

    Google Scholar 

  • Yazulla, S., Studholme, K. M., Mcintosh, H. H. & Fan, S. F. (2000) Cannabinoid receptors on goldfish retinal bipolar cells: Electron-microscope immunocytochemistry and whole-cell recordings. Visual Neuroscience 17, 391–401.

    Google Scholar 

  • Zygmunt, P. M., Petersson, J., Andersson, D. A., Chuang, H., Sorgard, M., Dimarzo, V., Julius, D. & Hogestatt, E. D. (1999) Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400, 452–457.

    Google Scholar 

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  1. Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, NY, 11794-5230, USA

    Sarah Zimov & Stephen Yazulla

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  1. Sarah Zimov
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Zimov, S., Yazulla, S. Localization of vanilloid receptor 1 (TRPV1/VR1)-like immunoreactivity in goldfish and zebrafish retinas: Restriction to photoreceptor synaptic ribbons. J Neurocytol 33, 441–452 (2004). https://doi.org/10.1023/B:NEUR.0000046574.72380.e8

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  • DOI: https://doi.org/10.1023/B:NEUR.0000046574.72380.e8

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