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Ultrastructural localisation of ATP-gated P2X2 receptor immunoreactivity in the rat hypothalamo-neurohypophysial system

  • Published:
Journal of Neurocytology

Abstract

The distribution of the P2X2 subtype of the purine receptor associated with the extracellular signalling activities of ATP was studied in the rat hypothalamo-neurohypophysial system at the electron microscope level. Receptors were labelled with ExtrAvidin-horseradish peroxidase preembedding immunocytochemistry using a polyclonal antibody against a fragment of an intracellular domain of the receptor. Immunoreactivity to P2X2 receptors was localised in: (i) paraventricular and supraoptic nuclei—in subpopulations of endocrine neurones, neurosecretory and non-neurosecretory axons and dendrites; and (ii) the neurohypophysis—in pituicytes and subpopulation of neurosecretory axons. In both the hypothalamic nuclei examined, labelled asymmetric axo-dendritic synapses were commonly observed. These synapses involved either P2X2-labelled axon terminals (synaptic buttons) and unlabelled dendrites or labelled dendrites and unlabelled axon terminals. Axo-somatic synapses established by P2X2-positive axons on P2X2-positive endocrine cell bodies as well as on P2X2-negative somata were also observed. The functional significance of these findings is discussed.

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References

  • Araque, A., Parpura, V., Sanzgiri, R. P. & Haydon, P. G. (1999) Tripartite synapses: glia, the unacknowledged partner. Trends in Neuroscience 22, 208-215.

    Google Scholar 

  • Bo, X., Zhang, Y., Nassar, M., Burnstock, G. & Schoepfer, R. (1995) A P2X purinoceptor cDNA conferring a novel pharmacological profile. FEBS Letters 375, 129-133.

    PubMed  Google Scholar 

  • Boersma, C. J., Sonnemans, M. A. & Van Leeuwen, F. M. (1993a) Immunoelectron microscopic demonstration of oxytocin and vasopressin in pituicytes and in nerve terminals forming synaptoid contacts with pituicytes in the rat neural lobe. Brain Research 611, 117-129.

    PubMed  Google Scholar 

  • Boersma, C. J., Van Leeuwen, F. W., Oõbrien, W. G., Law, G. J., Mason, W. T. & Bicknell, R. J. (1993b) Dynorphin 1-17 delays the vasopressin induced mobilization of intracellular calcium in cultured astrocytes from the rat neural lobe. Journal of Neuroendocrinology 5, 583-590.

    PubMed  Google Scholar 

  • Buller, K. M., Khanna, S., Sibbald, J. R. & Day, T. A. (1996) Central noradrenergic neurons signal via ATP to elicit vasopressin responses to haemorrhage Neuroscience 73, 637-642.

    PubMed  Google Scholar 

  • Chan, C. M., Unwin, R. J., Oglesby, I. B., Ford, A. P. D. W., Townsend-Nicholson, A. & Burnstock, G. (1998) Localization of the P2X1 purinoceptors by autoradiography and immunohistochemistry in rat kidneys. American Journal of Physiology 274, F799-F804.

    PubMed  Google Scholar 

  • Chen, Z. P., Levy, A. & Lightman, S. L. (1994) Activation of specific ATP receptors induces a rapid increase intracellular calcium ions in rat hypothalamic neurons. Brain Research 641, 249-256.

    PubMed  Google Scholar 

  • Chen, Z. P., Kratzmeier, M., Levy, A., Mcardle, C. A., Poch, A., Day, A., Mukhopathyay, A. K. & Lightman, S. L (1995) Evidence for a role of pituitary ATP receptors in the regulation of pituitary function. Proceedings of the National Academy of Science USA 92, 5219-5223.

    Google Scholar 

  • Collo, G., North, R. A., Kawashima, E., Merlo-Pich, E., Neidhart, S., Surprenant, A. & Buell, G. (1996) Cloning of P2X5 and P2X6 receptors and the distribution and properties of an extended family of ATP-gated ion channels. Journal of Neuroscience 16, 2495-2507.

    PubMed  Google Scholar 

  • Day, T. A., Sibbald, J. R. & Khana, S. (1993) ATP mediates an excitatory noradrenergic neurons input to supraoptic vasopressin cells. Brain Research 607, 341-344.

    PubMed  Google Scholar 

  • Gratzl, M., Torp-Pedersen, C., Daertt, D. A., Treiman, M. & Thorn, N. A. (1980) Isolation and characterization of secretory granules from bovine neurohypophyses. Hoppe-SeylerÕs Zeitschrift f ¬ur Physiologische Chemie 361, 1615-1628.

    Google Scholar 

  • Harboe, N. & Ingild, A. (1973) Immunization, isolation of immunoglobulins, estimation of antibody titre. Scandinavian Journal of Immunology 1 (Suppl), 161-164.

    PubMed  Google Scholar 

  • Hatton, G. I. (1990) Emerging concepts of structurefunction dynamics in adult brain; the hypothalamoneurohypophysial system. Progress of Neurobiology 34, 437-504.

    Google Scholar 

  • Hatton, G. I., Bunting, R. & Mason, W. (1992) Arginine vasopressin mobilises intracellular calcium via V1-receptor action in astrocytes (pituicytes) cultured from adult neural lobes. Brain Research 588, 75-83.

    PubMed  Google Scholar 

  • Hiruma, H. & Bourque, C. W., (1995) P2 purinoceptormediated depolarization of rat supraoptic neurosecretory cells in vitro. Journal of Physiology 489, 805-811.

    PubMed  Google Scholar 

  • Kidd, E. J., Grahames, C., Simon, J., Michel, A., Barnard, E. & Humphrey, P. P. A. (1995) Localization of P2X-purinoceptor transcripts in the rat nervous system. Molecular Pharmacology 48, 569-573.

    PubMed  Google Scholar 

  • Koshimizu, T., Tomic, M., Van Goor, F. & Stojilkovic, S. S. (1998) Functional role of alternative splicing in pituitary P2X2 receptor-channel activation and desensitization. Molecular Endocrinology 12, 901-913.

    PubMed  Google Scholar 

  • Loesch, A. & Burnstock, G. (1998) Electronimmunocytochemical localization of P2X1 receptors in the rat cerebellum. Cell and Tissue Research 294, 253-260.

    PubMed  Google Scholar 

  • Mori, M., Tsushima, H. & Matsuda, T. (1992) Antidiuretic effects of purinoceptor agonists injected into the hypothalamic paraventricular nucleus of waterloaded, ethanol-anesthetized rats, Neuropharmacology 31, 585-592.

    PubMed  Google Scholar 

  • Neary, J. T., Laskey, R., Van Breemen, C., Blicharska, J., Norenberg, L. O. & Norenberg, M. D. (1991) ATP-evoked calcium signal stimulates protein phosphorylation/dephosphorylation in astrocytes. Brain Research 566, 89-94.

    PubMed  Google Scholar 

  • Nordmann, J. J. & Chevallier, J. (1980) The role of microvesicles in buffering [Ca2+]I in the neurohypophysis. Nature 287, 54-56.

    PubMed  Google Scholar 

  • Potter, P. & White, T. D. (1980) Release of adenosine 50-triphosphate from synaptosomes from different regions of the brain. Neuroscience 5, 1351-1356.

    PubMed  Google Scholar 

  • Shaw, F. D. & Dyball, R. E. (1984) The relationship between calcium uptake and hormone release in the isolated neurohypophysis. A reassessment. Neuroendocrinology 38, 504-510.

    PubMed  Google Scholar 

  • Shaw, F. D. & Morris, J. F. (1980) Calcium localization in the rat neurohypophysis. Nature 287, 56-58.

    PubMed  Google Scholar 

  • Shibuya, I., Tanaka, K., Hattori, Y., Uezono, Y., Harayama, N., Noguchi, J., Ueta, Y., Izumi, F. & Ymashita, H. (1999) Evidence that multiple P2X2 purinoceptors are functionally expressed in rat supraoptic neurones. Journal of Physiology 514, 351-367.

    PubMed  Google Scholar 

  • Sperl´agh, B., Sershen, H., Lajtha, A. & Vizi, E. S. (1998a) Co-release of endogenous ATP and [3H]noradrenaline from rat hypothalamic slices: origin and modulation by ®2-adrenoceptors. Neuroscience 82, 511-520.

    PubMed  Google Scholar 

  • Sperl´agh, B., Mergl, Z S., Jur´anyi, ZS., Vizi, E. S. & Makara, G. B. (1998b) Local regulation of vasopressin and oxytocin secretion by extracellular ATP in the isolated posterior lobe of the rat hypophysis. Journal of Endocrinology 160, 343-360.

    Google Scholar 

  • Stoeckel, M. E., Hindelang-Gertner, C., Dellmann, H. D., Porte, A. & Stutinsky, F. (1975) Subcellular localization of calcium in the mouse hypophysis. I. Calcium distribution in the adenoand neurohypophysis under normal conditions. Cell and ssue Research 157, 307-322.

    Google Scholar 

  • Theodosis, D. T., El Majdoubi, M., Pierre, K. & POULAIN, D. A. (1998) Factors governing activitydependent structural plasticity of the hypothalamoneurohypophysial system. Cell and Molecular Neurobiology 18, 285-2

    Google Scholar 

  • Todorov, L. D., Mihaylova-Todorova, C. Craviso, G. L., Bjur, R. A. & Westfall, D. P. (1996) Evidence for the differential release of the cotransmitters ATP and noradrenaline from sympathetic nerves of the guinea-pig vas deferens. Journal of Physiology 496, 731-748.

    PubMed  Google Scholar 

  • Troadec, J. D., Thirion, S., Nicaise, G., Lemos, J. R. & Dayanith, G. (1998) ATP-evoked increases in [Ca2+]I and peptide release from rat isolated 504 LOESCH, MIAH and BURNSTOCK neurohypophysial terminals via a P2X2 purinoceptor. Journal of Physiology 511, 89-103.

    PubMed  Google Scholar 

  • Vulchanova, L., Arvidsson, U., Riedl, M., Wang, J., Buell, G., Surprenant, A., North, R. A. & Elde, R. (1996) Differential distribution of two ATP-gated ion channels (P2X receptors) determined by immunocytochemistry. Proceedings of the National Academy of Science USA 93, 8063-8067.

    Google Scholar 

  • Xiang, Z., Bo, X., Oglesby, I., Ford, A. & Burnstock, G. (1998) Localization ofATP-gatedP2X2 receptor immunoreactivity in the rat hypothalamus. Brain Research 813, 390-397.

    PubMed  Google Scholar 

  • Zimmermann, H. (1994) Signalling via ATP in the nervous system. Trends in Neurosciences 17, 420-426.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, UK

    Andrzej Loesch & Suhel Miah

  2. Autonomic Neuroscience Institute, Royal Free and University College Medical School, Rowland Hill Street, London, NW3 2PF, UK

    Geoffrey Burnstock

Authors
  1. Andrzej Loesch
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  2. Suhel Miah
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  3. Geoffrey Burnstock
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Loesch, A., Miah, S. & Burnstock, G. Ultrastructural localisation of ATP-gated P2X2 receptor immunoreactivity in the rat hypothalamo-neurohypophysial system. J Neurocytol 28, 495–504 (1999). https://doi.org/10.1023/A:1007009222518

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  • DOI: https://doi.org/10.1023/A:1007009222518

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