ABSTRACT
Aim ToconstructHEK293cellsstablyex-pressing corticotropin releasing factor receptor 1 ( CRFR1 ) , and evaluate its function by the cAMP as-say.Methods CulturedHEK293cellsweretransfect-ed with CRFR1-expressing vector by Lipofectamine 2000 and were selected by using G418 . CRFR1 ex-pression was detected by Western blot, RT-PCR and immunofluorescence.Results Westernblot,RT-PCR and immunofluorescence data revealed that the HEK293 cells expressed CRFR1 protein stably. The dose-responsive relationship experiment revealed that CRF induced a CRFR1-mediated cAMP production in HEK293 cells with EC50 =(5. 64 ± 0. 05) × 10 -10 mol ·L-1.Conclusion HEK293celllinesstablyex-pressing CRFR1 were constructed successfully, which would provide a cellular model to facilitate the research on the biological function of CRFR1 and CRFR1-targe-ted drug screening.
ABSTRACT
Stress exerts profound effects on cognitive and emotional functions. As a regulator of response to stress,the corticotropin releasing factor (CRF) during stress impacts organism function through hypothalamic-pituitary-adrenal axis. Recent research has shown that dendritic branches were abnormal and dendritic spine was lost under stress. In contrast,stressed mice with CRF receptor 1 (CRFR1) deficiency or CRFR1 antagonist exhibited normal dendritic morphology of hippocampal neurons .The structure and related factors expression of hippocampal neurons were affected by CRFR1 as a G protein-coupled receptor(GPCR), but the molecular mechanism about CRF-induced direct injury effect on hippocampal neurons has been poorly understood. The paper reviews the research progress on the contribution of stress activated CRF-CRF receptor signaling to the structure and related factors of hippocampal neurons.