Rotenone-induced changes of cysteinyl leukotriene receptor 1 expression in BV2 microglial cells.
- Author:
Jiang-Yun LUO
1
;
Zhuang ZHANG
;
Shu-Ying YU
;
Bing ZHAO
;
Chun-Zhen ZHAO
;
Xin-Xin WANG
;
San-Hua FANG
;
Wei-Ping ZHANG
;
Li-Hui ZHANG
;
Er-Qing WEI
;
Yun-Bi LU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cells, Cultured; Male; Mice; Microglia; drug effects; metabolism; pathology; Rabbits; Receptors, Leukotriene; immunology; metabolism; Rotenone; pharmacology
- From: Journal of Zhejiang University. Medical sciences 2011;40(2):131-138
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare and identify a polyclonal antibody (pAb) against (mouse) cysteinyl leukotriene receptor 1 (CysLT(1)) and to investigate the changes of CysLT(1) receptor expression in BV2 microglial cells after rotenone treatment.
METHODSRabbits were immunized with KLH-coupled CysLT(1) peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by ELISA method, and the specificity of the pAb was tested by antigen blockade. After BV2 cells were treated with rotenone (0.01-1 μmol/L) for 24 h, the expression of CysLT(1) was determined by immunostaining, Western blotting and RT-PCR.
RESULTThe pAb showed a titer of 1/32728, and was not cross-reacted with antigens of CysLT(2) receptor and GPR17. Immunostaining, Western blotting and RT-PCR analysis showed the expression of CysLT(1) receptor in BV2 microglia. Rotenone at 1μmol/L significantly induced an increased expression of CysLT(1) receptor.
CONCLUSIONThe prepared CysLT(1) receptor polyclonal antibody has a high titer and high specificity to meet testing requirements of Western blotting and immunostaining; CysLT(1) is associated with rotenone-induced injury of BV2 microglial cells.
