title: [Rotenone-induced changes of cysteinyl leukotriene receptor 1 expression in BV2 microglial cells].
abstract: To 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.
  
  
  
  Rabbits 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.
  
  
  
  The 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.
  
  
  
  The 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.
authors.affiliation: Department of Pharmacology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
authors.forename: Jiang-Yun
authors.initials: JY
authors.lastname: Luo
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Zhang
authors.forename: Shu-Ying
authors.initials: SY
authors.lastname: Yu
authors.forename: Bing
authors.initials: B
authors.lastname: Zhao
authors.forename: Chun-Zhen
authors.initials: CZ
authors.lastname: Zhao
authors.forename: Xin-Xin
authors.initials: XX
authors.lastname: Wang
authors.forename: San-Hua
authors.initials: SH
authors.lastname: Fang
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Zhang
authors.forename: Li-Hui
authors.initials: LH
authors.lastname: Zhang
authors.forename: Er-Qing
authors.initials: EQ
authors.lastname: Wei
authors.forename: Yun-Bi
authors.initials: YB
authors.lastname: Lu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D008297:Male; D051379:Mice; D017628:Microglia / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D011817:Rabbits; D018077:Receptors, Leukotriene / Q000276:immunology / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology*
chemical_list: D018077:Receptors, Leukotriene; D012402:Rotenone; C083169:leukotriene D4 receptor