Katai Naomichi, Yanagidaira Tomoko, Senda Nami, Murata Toshinori, Yoshimura Nagahisa
Department of Ophthalmology, Shinshu University School of Medicine, Matsumoto, Japan.
Jpn J Ophthalmol. 2006 Mar-Apr;50(2):121-7. doi: 10.1007/s10384-005-0296-7.
To determine if c-Jun and Bcl-2 family proteins play a role in photoreceptor apoptosis in Royal College of Surgeons (RCS) rats.
RCS and Sprague-Dawley rats were used. Cryosections of retinas harvested at various postnatal periods were immunostained with antibodies against c-Jun, Bcl-2, and Bax. Double staining with TdT-dUTP nick-end labeling (TUNEL) or propidium iodide (PI) and antibodies was also done. To study the time course of gene and protein expression, semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting analyses were carried out.
TUNEL-positive photoreceptors of RCS rats were stained strongly with antibodies against c-Jun and Bax. The number of immunoreactive cells increased on days 21 and 28 after birth (P21 and P28) and decreased on P45. Semiquantitative RT-PCR analysis showed that mRNAs for c-Jun and Bax were upregulated at P21 and P28, but those for Bcl-2 were unchanged. On immunoblotting, a 43-kDa band was revealed by the anti-c-Jun antibody and a 21-kDa band, by the anti-Bax antibody. Protein expression of c-Jun and Bax were increased at both P21 and P28. The temporal profiles of immunoreactivity, protein expression, and mRNA expression were similar.
c-Jun and Bax may play a role in photoreceptor apoptosis in RCS rats.
确定c-Jun和Bcl-2家族蛋白是否在皇家外科学院（RCS）大鼠的光感受器细胞凋亡中发挥作用。
使用RCS大鼠和Sprague-Dawley大鼠。对在不同出生后时期采集的视网膜冰冻切片用抗c-Jun、Bcl-2和Bax的抗体进行免疫染色。还进行了TdT-dUTP缺口末端标记（TUNEL）或碘化丙啶（PI）与抗体的双重染色。为了研究基因和蛋白表达的时间进程，进行了半定量逆转录聚合酶链反应（RT-PCR）和免疫印迹分析。
RCS大鼠的TUNEL阳性光感受器被抗c-Jun和Bax抗体强烈染色。免疫反应性细胞数量在出生后第21天和第28天（P21和P28）增加，在P45时减少。半定量RT-PCR分析显示，c-Jun和Bax的mRNA在P21和P28时上调，但Bcl-2的mRNA未改变。在免疫印迹中，抗c-Jun抗体显示出一条43-kDa的条带，抗Bax抗体显示出一条21-kDa的条带。c-Jun和Bax的蛋白表达在P21和P28时均增加。免疫反应性、蛋白表达和mRNA表达的时间变化趋势相似。
c-Jun和Bax可能在RCS大鼠的光感受器细胞凋亡中发挥作用。
