title: Transgenic approach to express the channelrhodopsin 2 gene in arginine vasopressin neurons of rats.
abstract: Optogenetics provides a powerful tool to regulate neuronal activity by light-sensitive ion channels such as channelrhodopsin 2 (ChR2). Arginine vasopressin (AVP; also known as the anti-diuretic hormone) is a multifunctional hormone which is synthesized in the magnocellular neurosecretory cells (MNCs) of the hypothalamus. Here, we have generated a transgenic rat that expresses an AVP-ChR2-enhanced green fluorescent protein (eGFP) fusion gene in the MNCs of the hypothalamus. The eGFP fluorescence that indicates the expression of ChR2-eGFP was observed in the supraoptic nucleus (SON) and in the magnocellular division of the paraventricular nucleus (PVN) that is known to contain AVP-secreting neurons. The eGFP fluorescence intensities in those nuclei and posterior pituitary were markedly increased after chronic salt loading (2% NaCl in drinking water for 5days). ChR2-eGFP was localized mainly in the membrane of AVP-positive MNCs. Whole-cell patch-clamp recordings were performed from single MNCs isolated from the SON of the transgenic rats, and blue light evoked repetitive action potentials. Our work provides for the first time an optogenetic approach to selectively activate AVP neurons in the rat.
authors: Department of Physiology, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan; Department of Pediatrics, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Masahiro
authors: M
authors: Ishii
authors: Department of Physiology, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Hirofumi
authors: H
authors: Hashimoto
authors: Department of Otorhinolaryngology-Head and Neck Surgery, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Jun-Ichi
authors: JI
authors: Ohkubo
authors: Department of Otorhinolaryngology-Head and Neck Surgery, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Toyoaki
authors: T
authors: Ohbuchi
authors: Department of Neurosurgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Takeshi
authors: T
authors: Saito
authors: Department of Physiology, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Takashi
authors: T
authors: Maruyama
authors: Department of Physiology, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Mitsuhiro
authors: M
authors: Yoshimura
authors: Department of Pediatrics, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Yukiyo
authors: Y
authors: Yamamoto
authors: Department of Pediatrics, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan.
authors: Koichi
authors: K
authors: Kusuhara
authors: Department of Physiology, University of Occupational and Environmental Health, Kitakyushu, 807-8555 Japan. Electronic address: yoichi@med.uoeh-u.ac.jp.
authors: Yoichi
authors: Y
authors: Ueta
mesh_terms: D000818:Animals; D001127:Arginine Vasopressin / Q000235:genetics / Q000378:metabolism*; D000075402:Channelrhodopsins; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D008297:Male; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D062308:Optogenetics / Q000379:methods*; D010286:Paraventricular Hypothalamic Nucleus / Q000378:metabolism*; D010904:Pituitary Gland, Posterior / Q000378:metabolism; D051381:Rats; D055647:Rats, Transgenic; D017208:Rats, Wistar; D012965:Sodium Chloride / Q000008:administration & dosage; D013495:Supraoptic Nucleus / Q000378:metabolism* / Q000502:physiology
chemical_list: D000075402:Channelrhodopsins; C485184:enhanced green fluorescent protein; D001127:Arginine Vasopressin; D049452:Green Fluorescent Proteins; D012965:Sodium Chloride
keywords: Arginine vasopressin; Channelrhodopsin 2; Rat; Salt loading; Transgene; Whole-cell patch-clamp recording