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Hiroki Otsubo Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan
Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Susumu Hyodo Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Hirofumi Hashimoto Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Makoto Kawasaki Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Hitoshi Suzuki Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Takeshi Saito Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Toyoaki Ohbuchi Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Toru Yokoyama Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Hiroaki Fujihara Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Tetsuro Matsumoto Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Yoshio Takei Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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Yoichi Ueta Department of Physiology, Laboratory of Physiology, Department of Urology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan

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We examined the effects of i.c.v. administration of adrenomedullin 5 (AM5) on the brain of conscious rats. We used porcine AM5 in the present study because rat AM5 has not been detected. We observed Fos-like immunoreactivity (LI) in the hypothalamus and brainstem of conscious rats after i.c.v. administration of AM5 (2 nmol/rat). Fos-LI, measured at 90 min post-AM5 injection, was observed in various brain areas, including the supraoptic (SON) and the paraventricular nuclei (PVN). Dual immunostaining for Fos/oxytocin (OXT) and Fos/arginine vasopressin (AVP) revealed that OXT-LI neurones predominantly colocalized Fos-LI compared with AVP-LI neurones in the SON and the PVN. Plasma OXT levels were significantly increased 5 min after i.c.v. administration of AM5 (1 nmol/rat) compared with vehicle and remained elevated in samples taken at 15 and 30 min without changes in plasma AVP levels at any time. In situ hybridization histochemistry showed that i.c.v. administration of AM5 (0.2, 1 and 2 nmol/rat) caused a marked induction of the expression of the c-fos gene in the SON and the PVN. This induction was significantly but not completely reduced by pretreatment with both the calcitonin gene-related peptide (CGRP) antagonist CGRP-(8–37; 3 nmol/rat) and the AM receptor antagonist AM-(22–52; 27 nmol/rat). Although porcine AM5 has not been detected yet in the brain, these results suggest that centrally administered porcine AM5 may activate OXT-secreting neurosecretory cells in the hypothalamus partly through AM/CGRP receptors and elicit secretion of OXT into the systemic circulation in conscious rats.

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Akiko Katoh Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan
Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Hiroaki Fujihara Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Toyoaki Ohbuchi Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan
Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Tatsushi Onaka Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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W Scott Young III Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Govindan Dayanithi Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Yuka Yamasaki Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Mitsuhiro Kawata Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Hitoshi Suzuki Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Hiroki Otsubo Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Hideaki Suzuki Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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David Murphy Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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Yoichi Ueta Departments of, Physiology, Otorhynolaryngology, Department of Physiology, Section on Neural Gene Expression, Department of Cellular Neurophysiology, Department of Anatomy and Neurobiology, Molecular Neuroendocrinology Research Group, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan

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We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the OxteCfp fusion gene was expressed in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence and the posterior pituitary (PP). In in vitro preparations, freshly dissociated cells from the SON and axon terminals showed clear eCFP fluorescence. Immunohistochemistry for OXT and arginine vasopressin (AVP) revealed that the eCFP fluorescence co-localises with OXT immunofluorescence, but not with AVP immunofluorescence in the SON and the PVN. Although the expression levels of the OxteCfp fusion gene in the SON and the PVN showed a wide range of variations in transgenic rats, eCFP fluorescence was markedly increased in the SON and the PVN, but decreased in the PP after chronic salt loading. The expression of the Oxt gene was significantly increased in the SON and the PVN after chronic salt loading in both non-transgenic and transgenic rats. Compared with wild-type animals, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration and OXT and AVP levels, suggesting that the fusion gene expression did not disturb any physiological processes. These results suggest that our new transgenic rats are a valuable new tool to identify OXT-producing neurones and their terminals.

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