新しい脳ペプチド分離の新戦略

2001 年度 実績報告書

• 研究課題/領域番号: 11354010
• 研究機関: 国立遺伝学研究所
• 研究代表者: 藤澤 敏孝 国立遺伝学研究所, 個体遺伝研究系, 助教授 (60000262)
• 研究分担者: 筒井 和義 広島大学, 総合科学部, 教授 (20163842) ; 小泉 修 福岡女子大学, 人間環境学部, 教授 (50094777) ; 岡戸 信男 筑波大学, 基礎医学系, 教授 (50060140) ; 松島 治 広島大学, 理学部, 教授 (80127711) ; 藤本 正昭 島根大学, 生物資源科学部, 教授 (90081728)
• キーワード: 脳ペプチド / ラット / トリ / 両生類 / 無脊椎動物

研究概要

1.ヒドラペプチドの組織的単離・同定:360のペプチドを単離し、内122種についてアミノ酸配列を決定し、38種を化学合成し生物活性検定に供した。最終的には新規神経ペプチド6種、上皮ペプチド5種を同定し、これらをコードする遺伝子全てを同定した。また、抗ペプチド抗体の作成、生物活性検定を順調にこなし、非常に興味あるペプチド分子を多く得ることが出来た。
2.抗PKVamide(Hym-176)抗体を用いたラット脳の免疫組織染色:我々は既に抗GLWamide抗体を用いてラット脳の免疫組織染色を行い、抗GLWamide抗体陽性細胞が腹側被蓋野に存在することを観察している。同様に、抗PKVamide抗体陽性細胞が視床下部弓状核腹側の外足部に観察された。これらのことからヒドラの神経ペプチドの類似ペプチドが脊椎動物の脳にも存在すると予想された。
3.抗ペプチド抗体3種(抗RFamide、抗GLWamide、抗PKVamide)を利用した脳ペプチドの分離:ヒドラを含む腔腸動物の神経ペプチド抗体を利用し、ELISA法を用い脊椎動物の脳から抗体と反応するペプチドの単離を行った。抗RFamide抗体を用いて、鳥類脳の視床下部からRFamideファミリーに属する新規のゴナドトロピン放出抑制ペプチドを同定した。両生類からもC-末端がRFamideの成長ホルモン放出ペプチドを同定した。また、ラット脳から抗GLWamide抗体と反応するペプチドを既に同定したが、抗PKVamide抗体と反応するペプチドも単離した。
4.その他の脳ペプチド:マダコ脳からマダコ循環系に作用するペプチド2種、生殖系に作用するペプチド3種を得た。またフナ脳から同定したC-RFa(mide)は視床下部で産生され、下垂体でプロラクチン(PRL)放出を促進し、PRLは血流で標的細胞(塩細胞と考えられる)に達し、浸透圧調整を行うと予想された。

研究成果

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