ペプチドーム解析を応用した脳機能調節に作用する新規神経ペプチドの探索と機能研究

2006 Fiscal Year Annual Research Report

• Project/Area Number: 18022035
• Research Institution: University of Miyazaki
• Principal Investigator: 中里 雅光 宮崎大学, 医学部, 教授 (10180267)
• Co-Investigator(Kenkyū-buntansha): 塩見 一剛 宮崎大学, 医学部, 助教授 (40305082) ; 十枝内 厚次 宮崎大学, 医学部, 助手 (80381101) ; モンダール M・S 宮崎大学, 医学部, 研究生 (00398265) ; 新原 琢也 宮崎大学, 医学部, 助手 (10398266) ; 松尾 崇 宮崎大学, 医学部, 医員 (10404427)
• Keywords: 摂食・エネルギー代謝調節 / ペプチドの網羅的解析 / オーファン受容体 / Ca^<2+>イメージング装置 / 腫瘍細胞株 / 高感度RIA系 / NERP / vgf

Research Abstract

培養細胞からのペプチドーム・ペプチドミクスを活用したペプチドの網羅的解析を行えば、前駆体蛋白由来のペプチドフラグメントが多数同定でき、その前駆体蛋白の遺伝1青報や新たに同定したペプチドフラグメントのアミノ酸配列から生理活性を有するペプチドが推定可能であると考えられた。腫瘍細胞株、オーファン受容体発現系の細胞株やラットの視床下部ニューロンから分離・培養した初代細胞、および96サンプルを一度に測定できる現有の最新Ca^<2+>イメージング装置(IMACS細胞内イオン動態画像解析システム)を用いて解析した結果、神経内分泌細胞株から細胞内カルシウム上昇活性を有する2つの生理活性ペプチド(NERP-1とNERP-2)を同定した。同定したペプチドのアミノ酸配列に基づき、ラット組織よりペプチドをコードするcDNA配列を決定し、ペプチド前駆体構造がNGFで誘導されるvgfであることを明らかにした。遺伝子操作技術、行動薬理学などに関するペプチドの網羅的解析法を構築し、NERPsの生理学的意義を明らかにしている。家兎に免疫して新規ペプチド特異的抗体を作製し、高感度RIA系を開発した。この定量系を用いて、NERPsの脳内濃度を検討した結果、海馬と視床下部に高い免疫活性を認めた。さらに細胞内情報伝達系の動態をhigh-through putで評価できるcell-based assay系を確立し、新規の神経ペプチドを4種類同定し、現在その機能解析を進めている。本申請での網羅的ペプチド解析で生理活性を有する新規ペプチドが同定されることが証明された。

Research Products

• [Journal Article] Ghrelin stimulates growth hormone secretion and food intake in aged rats. (2007)
  • Author(s): Toshinai K, Mondal MS, Shimbara T, Yamaguchi H, Date Y, Kangawa K, Nakazato M
  • Journal Title: Mech Ageing Dev 128 Pages: 715-721
  • Peer Reviewed
• [Journal Article] Cardiovascular actions of central neuropeptide W in conscious rats. (2007)
  • Author(s): Yu N, Chu C, Kunitake T, Kato K, Nakazato M, Kannan H
  • Journal Title: Regul Pept. 138 Pages: 82-86
• [Journal Article] Peripheral ghrelin transmits orexigenic signals through the noradrenergic pathway from the hindbrain to the hypothalamus. (2006)
  • Author(s): Date Y, Shimbara T, Koda S, Toshinai K, Ida T, Murakami N, Miyazato M, Kokame K, Ishizuka Y, Ishida Y, Kageyama H, Shioda S, Kangawa K, Nakazato M
  • Journal Title: Cell Metab 4 Pages: 323-331
• [Journal Article] Des-acyl ghrelin induces food intake by a mechanism independent of the growth hormone secretagogue receptor. (2006)
  • Author(s): Toshinai K, Yamaguchi H, Sun Y, Smith RG, Yamanaka A, Sakurai T, Date Y, Mondal MS, Shimbara T, Kawagoe T, Murakami N, Miyazato M, Kangawa K, Nakazato M
  • Journal Title: Endocrinology 147 Pages: 2306-2314
• [Journal Article] Central effects of CRSP-1 on energy homeostasis in rats. (2006)
  • Author(s): Sawada H, Yamaguchi H, Shimbara T, Toshinai K, Mondal MS, Date Y, Murakami N, Katafuchi T, Minamino N, Nunoi H, Nakazato M
  • Journal Title: Endocrinology 147 Pages: 2043-2050
• [Journal Article] Neuronal pathway from the liver modulates energy expenditure and systemic insulin sensitivity. (2006)
  • Author(s): Uno K, Katagiri H, Yamada T, Ishigaki Y, Ogihara T, Imai J, Hasegawa Y, Gao J, Kaneko K, Iwasaki H, Ishihara H, Sasano H, Inukai K, Mizuguchi H, Asano T, Shiota M, Nakazato M, Oka Y
  • Journal Title: Science 312 Pages: 1656-1659
• [Journal Article] Neuropeptide W is present in antral G cells of rat, mouse, and human stomach. (2006)
  • Author(s): Mondal MS, Kawagoe T, Yamaguchi H, Date Y, Kawagoe T, Tsuruta T, Kageyama H, Kawamura Y, Shioda S, Shimomura Y, Mori M, Nakazato M
  • Journal Title: J Endocrinol 188 Pages: 49-57
• [Journal Article] Galanin-like peptide promotes feeding behavior via activation of orexinergic neurons in the rat lateral hypothalamus. (2006)
  • Author(s): Kageyama H, Kita T, Toshinai K, Guan JL, Date Y, Takenoya F, Kato S, Ohtaki T, Matsumoto H, Nakazato M, Shioda S
  • Journal Title: J Neuroendocrinol 18 Pages: 33-41
• [Journal Article] Maternal ghrelin plays an important role in rat fetal development during pregnancy. (2006)
  • Author(s): Nakahara K, Nakagawa M, Baba Y, Sato M, Toshinai K, Date Y, Nakazato M, Kojima M, Miyazato M, Kaiya H, Hosoda H, Kangawa K, Murakami N
  • Journal Title: Endocrinology 147 Pages: 1333-1342
• [Journal Article] Postnatal development of orexin-A and orexin-B like immunoreactivities in the eastern grey kangaroo (macropus giganteus) hypothalamus. (2006)
  • Author(s): Yamamoto Y, Mckinley MJ, Nakazato M, Yamashita H, Shirahata A, Ueta Y
  • Journal Title: Neurosci Lett 392 Pages: 124-128
• [Journal Article] Fasting-induced reduction in locomotor activity and reduced response of orexin neurons in carnitine-deficient mice. (2006)
  • Author(s): Yoshida G, Li MX, Horiuchi M, Nakagawa S, Sakata M, Kuchiiwa S, Kuchiiwa T, Jalil MA, Begum L, Lu YB, Iijima M, Hanada T, Nakazato M, Huang ZL, Eguchi N, Kobayashi K, Saheki T
  • Journal Title: Neurosci Res 55 Pages: 78-86
• [Journal Article] Ghrelin concentrations and cardiac vagal tone are decreased after pharmacologic and cognitive-behavioral treatment in patients with bulimia nervosa. (2006)
  • Author(s): Tanaka M, Nakahara T, Muranaga T, Kojima S, Yasuhara D, Ueno H, Nakazato M, Inui A
  • Journal Title: Horm Behav 50 Pages: 261-265
• [Journal Article] Centrally administered neuropeptide W-30 activates magnocellular neurosecretory cells in the supraoptic and paraventricular nuclei with neurosecretion in rats. (2006)
  • Author(s): Kawasaki M, Onaka T, Nakazato M, Saito J, Mera T, Hashimoto H, Fujihara H, Okimoto N, Ohnishi H, Nakamura T, Ueta Y
  • Journal Title: J Endocrinol 190 Pages: 213-223
• [Journal Article] Changes in plasma ghrelin levels, gastric ghrelin production, and body weight after Helicobacter pylori cure. (2006)
  • Author(s): Osawa H, Kita H, Ohnishi H, Nakazato M, Date Y, Bowlus CL, Ishino Y, Watanabe E, Shiiya T, Ueno H, Hoshino H, Satoh K, Sugano K
  • Journal Title: J Gastroenterol 41 Pages: 954-961
• [Journal Article] Effects of ghrelin and des-acyl ghrelin on neurogenesis of the rat fetal spinal cord. (2006)
  • Author(s): Sato M, Nakahara K, Goto S, Kaiya H, Miyazato M, Date Y, Nakazato M, Kangawa K, Murakami N
  • Journal Title: Biochem Biophys Res Commun 350 Pages: 598-603
• [Journal Article] Incomplete restoration of the secretion of ghrelin and PYY compared to insulin after food ingestion following weight gain in anorexia nervosa.
  • Author(s): Nakahara T, Kojima S, Tanaka M, Yasuhara D, Harada T, Sagiyama KI, Muranaga T, Nagai N, Nakazato M, Nozoe SI, Naruo T, Inui A
  • Journal Title: J Psychiatr Res (in press)
• [Journal Article] Ontogeny of a New Enteric Peptide, Neuropeptide W (NPW), in the Developing Rat Stomach.
  • Author(s): Mondal MS, Yamaguchi H, Date Y, Tsuruta T, Shimbara T, Toshinai K, Shimomura Y, Mori M, Nakazato M
  • Journal Title: Regul Pept. (in press)
• [Journal Article] The role of PYY3-36 in the feeding regulation.
  • Author(s): Ueno H, Yamaguchi H, Mizuta M, Nakazato M
  • Journal Title: Regul Pept. (in press)