Wu Yulong, Cheng Mei, Shi Zhen, Feng Zhenqing, Guan Xiaohong
Key Lab of Antibody Technique of Health Ministry, Nanjing Medical University Nanjing, Jiangsu, P.R. China ; School of Basic Medical Science, Binzhou Medical University Binzhou, Shandong, P.R. China.
School of Nursing, Binzhou Medical University Binzhou, Shandong, P.R. China.
Int J Clin Exp Pathol. 2014 Dec 1;7(12):8563-72. eCollection 2014.
Pancreata from Sprague Dawley rats of different developmental stages were studied to determine the expression and cellular localization of different c-MET isoforms in the developing rat pancreas. Pancreatic mRNA and protein expression levels of c-MET at different developmental stages from embryo to adult were detected by reverse transcription-polymerase chain reaction and by western blotting. To identify the cellular localization of c-MET protein in the developing rat pancreas, double immunofluorescent staining was performed using antibodies for cell type-specific markers and for c-MET. The expression of two isoforms of c-MET (190 kDa and 170 kDa) coincided with the development of the pancreas. The 190 kDa isoform of c-MET is expressed during embryonic stages, and its expression is replaced by the expression of the 170 kDa isoform as the pancreas develops. Only the 170 kDa isoform is expressed in the adult rat pancreas. Throughout all stages of pancreatic development, c-MET is expressed by vimentin-positive cells. In contrast, c-MET staining was stronger in rat pancreata from newborn to adult stages and overlapped with insulin-positive beta-cells. The dynamic expression and localization of different c-MET isoforms in the rat pancreas during different developmental stages indicates that distinct c-MET isoform might be involved in different aspects of pancreatic development.
研究了不同发育阶段的斯普拉格-道利大鼠的胰腺,以确定不同c-MET异构体在发育中的大鼠胰腺中的表达和细胞定位。通过逆转录-聚合酶链反应和蛋白质印迹法检测从胚胎到成年不同发育阶段胰腺中c-MET的mRNA和蛋白质表达水平。为了确定c-MET蛋白在发育中的大鼠胰腺中的细胞定位,使用针对细胞类型特异性标志物和c-MET的抗体进行双重免疫荧光染色。c-MET的两种异构体(190 kDa和170 kDa)的表达与胰腺的发育一致。c-MET的190 kDa异构体在胚胎阶段表达,随着胰腺的发育,其表达被170 kDa异构体的表达所取代。成年大鼠胰腺中仅表达170 kDa异构体。在胰腺发育的所有阶段,波形蛋白阳性细胞表达c-MET。相比之下,从新生到成年阶段,大鼠胰腺中的c-MET染色更强,且与胰岛素阳性的β细胞重叠。不同发育阶段大鼠胰腺中不同c-MET异构体的动态表达和定位表明,不同的c-MET异构体可能参与胰腺发育的不同方面。