血清ショックにより,哺乳類の組織培養細胞における昼夜間の遺伝子発現が誘発される
A Balsalobre1, F Damiola, U Schibler
1Département de Biologie Moléculaire, Sciences II, Université de Genève, Switzerland.
Cell
|July 11, 1998
まとめ
血清で治療された培養細胞は,生体内リズムを反映した昼夜間の遺伝子発現を示します. この血清ショック法は,日中時計の遺伝子調節を研究する新しい方法を提供します.
科学分野:
- 分子生物学は分子生物学である.
- クロノバイオロジー クロノバイオロジー
- 細胞生物学 細胞生物学
背景:
- シルカディアンリズムとは,24時間のサイクルで生理学的機能を調節する内生的な生物学的プロセスである.
- 遺伝子転写は昼夜リズムの主な構成要素であり,多くの遺伝子はリズム的な発現パターンを表しています.
- 昼間の遺伝子発現の基礎にある分子機構を理解することは,生物学的タイミングの解読に不可欠です.
研究 の 目的:
- 培養細胞の血清治療が昼間の遺伝子発現を誘発できるかどうかを調査する.
- 培養細胞における遺伝子発現の時間的なパターンを,in vivoで観察されたものと比較する.
- サームショックの可能性を研究し,細胞内で昼夜リズムを研究するツールとして活用する.
主な方法:
- 培養されたラット-1線維芽細胞とH35肝腫細胞は,高濃度の血清で治療されました.
- rper1,rper2,Rev-Erb alpha,DBP,TEFを含む特定の遺伝子のmRNAレベルは,時間とともに定量化されました.
- 周期長と振動の数を培養細胞で分析した.
- 血清ショック後のc-fosとrper発現をモニタリングした.
主要な成果:
- 血清治療は,ドロソフィラ期遺伝子 (rper1とrper2) のラットホモログ,および転写因子Rev-Erbα,DBP,TEFを含む,培養細胞における複数の遺伝子の昼夜発現を誘導した.
- ラット-1の線維芽細胞は,平均22.5時間の連続した1日3回の振動を示した.
- 培養細胞におけるmRNA蓄積サイクルの時序は,in vivo観測と一致していた.
- 血清ショックは一時的にc-fosとrperの発現を刺激し,光誘発の即時早期の遺伝子発現を模倣した.
結論:
- 血清治療は,培養細胞の昼間遺伝子発現を誘発するのに有効です.
- 培養細胞は,昼夜リズムの分子メカニズムを研究するための貴重なモデルを提供します.
- 血清ショックメソッドは,細胞内での昼夜時計の遺伝子調節を調査するための新しいアプローチを提供し,潜在的に光誘発反応を模倣します.
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