Chung H O, Kato T, Kato Y
Biosignal Research Center, Gunma University, Japan.
Mol Cell Endocrinol. 1996 May 17;119(1):75-82. doi: 10.1016/0303-7207(96)03798-7.
The presence of the typical transcription factors c-Jun, c-Fos and cAMP-responsive element (CRE)-binding protein in the porcine anterior pituitary was examined by molecular cloning and their involvement in the membrane signal cascade, especially their roles in gonadotropin-releasing hormone (GnRH) stimulation, were studied. Several cDNA clones were isolated from a porcine anterior pituitary cDNA library using cDNA probes. They were identified as porcine c-jun and c-fos by determining their nucleotide sequences, but a homologue for CREB341 which is a member of CRE-binding protein was not detected in porcine anterior pituitary. Reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to estimate the c-jun and c-fos mRNA contents in GnRH-, forskolin- (cAMP activator) and tetradecanoyl phorbol acetate- (TPA; protein kinase C activator) treated primary cultures of porcine anterior pituitary cells. Densitometric quantification demonstrated that GnRH and TPA treatment increased c-jun and c-fos mRNA levels significantly, whereas forskolin reduced the levels of both. Therefore, c-Jun and c-Fos are definitely present in porcine anterior pituitary and their mRNAs differentially involved in the signal transduction pathway mediated by two kinases. In particular, GnRH might regulate gonadotropin expression by increasing of c-jun and c-fos levels.
通过分子克隆技术检测了猪垂体前叶中典型转录因子c-Jun、c-Fos和环磷酸腺苷反应元件(CRE)结合蛋白的存在情况,并研究了它们在膜信号级联反应中的作用,特别是它们在促性腺激素释放激素(GnRH)刺激中的作用。使用cDNA探针从猪垂体前叶cDNA文库中分离出几个cDNA克隆。通过测定其核苷酸序列,将它们鉴定为猪c-jun和c-fos,但在猪垂体前叶中未检测到作为CRE结合蛋白成员的CREB341的同源物。进行逆转录-聚合酶链反应(RT-PCR)分析,以估计GnRH、福斯高林(cAMP激活剂)和十四酰佛波醇乙酸酯(TPA;蛋白激酶C激活剂)处理的猪垂体前叶细胞原代培养物中c-jun和c-fos mRNA的含量。光密度定量分析表明,GnRH和TPA处理显著增加了c-jun和c-fos mRNA水平,而福斯高林降低了两者的水平。因此,c-Jun和c-Fos确实存在于猪垂体前叶中,它们的mRNA在由两种激酶介导的信号转导途径中发挥不同作用。特别是,GnRH可能通过增加c-jun和c-fos水平来调节促性腺激素的表达。