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Transgenic Expression of Enhanced Green Fluorescent Protein Enables Direct Visualization for Physiological Studies of Vasopressin Neurons and Isolated Nerve Terminals of the Rat

Department of Physiology (Y.U., H.F., R.S.), School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan; U-583 Institut National de la Santé et de la Recherche Médicale (G.D.), L’Institut des Neurosciences de Montpellier, Hopital Saint Eloi, Montpellier F-34091, France; Department of Anatomy and Neurobiology (H.O., K.-i.M., M.K.), Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan; Department of Biological Information Processing (J.Y., A.F.), Graduate School of Electronic Science and Technology, Shizuoka University, Hamamatsu 432-8011, Japan; Department of Physiology (S.U., A.F.), Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan; and Molecular Neuroendocrinology Research Group (D.M.), The Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Bristol BS1 3NY, United Kingdom

Address all correspondence and requests for reprints to: Yoichi Ueta, M.D., Ph.D., Department of Physiology, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan. E-mail: yoichi{at}med.uoeh-u.ac.jp.

We have generated transgenic rats expressing an arginine vasopressin (AVP)-enhanced green fluorescent protein (eGFP) fusion gene. The expression of the eGFP gene and strong fluorescence were observed in the supraoptic nucleus (SON), the paraventricular nucleus (PVN), and the suprachiasmatic nucleus (SCN) in transgenic rats. The hypothalamo-neurohypophyseal tract, isolated SON neurons, and isolated axon terminals in the neurohypophysis also showed robust eGFP fluorescence. Water deprivation for 2 d increased the fluorescence of the eGFP in the SON and the PVN but not the SCN. The whole-cell patch-clamp technique was then used to record the electrical activities specifically identifying eGFP-expressing SON, PVN, and SCN AVP neurons in in vitro brain slice preparations. The AVP-eGFP transgenic rats are a unique new tool with which to study the physiological role of AVP-secreting neurons in the central nervous system and the dynamics of the regulation of AVP secretion in the living neurons and their axon terminals.

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