国际神经病学神经外科学杂志2011，Vol.38Issue(1)：19-23,5.
亨廷顿蛋白相关蛋白1与脑源性神经营养因子的胞吞关系的机制研究
Huntingtin associated protein 1 is involved with the endocytosis of mature brain derived neurotrophic factor
陈锶 1肖波 1毕方方 1冯莉1
作者信息
- 1. 中南大学湘雅医院神经内科,湖南省长沙市,410008
- 折叠
摘要
Abstract
Objective To investigate the roles of Huntingtin associated protein 1 ( HAP1 ) in the endocytosis of mature brain derived neurotrophic factor (mBDNF) and the underlying mechanism. Methods PC12 cells were differentiated by NGF and co-transfected with plasmid of HAP1 A-CFP and (or) mBDNF-ds-red. The cells were incubated with either recombinant ds-red-labeled mBDNF, or in combination of sheep anti-BDNF antibodies or rabbit anti-p75NTR. The expression of fluorescence and its intercellular location were determined by laser scanning confocal microscope. Cortical neurons from HAP1 +/+ and HAP1-/- mice at postnatal day 1 were cultured and treated with biotin labeled mBDNF to trigger endocytosis. After 60 minutes, the cells were washed and fixed, followed by immunostaining and confocal imaging. Results Co-transfected PC12 cells expressed partly co-localization of HAPl with mBDNF. Cells transfected with HAPl A-CFP which were incubated with recombinant ds-red-labeled mBDNF showed almost complete co-localization of HAPl with mBDNF. The antibodies to BDNF and p75NTR abolished co-internalization of HAPl with mBDNF. In addition, the labeled mBDNF was detected in almost all HAP1+/+, but not in HAP1-/- cortical neurons. Conclusions HAP1 plays an important role in the endocytosis of mBDNF.关键词
脑源性神经营养因子/亨廷顿蛋白相关蛋白1/转染/荧光质粒/激光共聚焦显微镜Key words
mature brain derived neurotrophic factor/ Huntingtin associated protein 1/ transfection/ fluorescence plasmids/ confocal microscope引用本文复制引用
陈锶,肖波,毕方方,冯莉..亨廷顿蛋白相关蛋白1与脑源性神经营养因子的胞吞关系的机制研究[J].国际神经病学神经外科学杂志,2011,38(1):19-23,5.
