title: [The wild-type alpha-synuclein over-expression to induce the protein aberrant aggregation of alpha-synuclein in HEK293 cells in vitro].
abstract: OBJECTIVE
  To investigate over-expression of wild-type alpha-synuclein inducing the aberrant aggregation of alpha-synuclein in HEK293 cell in vitro.
  
  
  METHODS
  The cDNA encoding the human alpha-synuclein without the stop code was cloned into PGEM T-easy vector. Using enzyme map and DNA sequencing analyzed and determined the recombinant plasmid, and then sub-clone the alpha-synuclein cDNA fragment into pEGFP-N1 vector. The recombinant plasmids alpha-synuclein-pEGFP were transfected into HEK293 cells by lipofectamin 2000. The aberrant aggregation of alpha-synuclein was measured by EGFP fluorescence, anti-alpha-synuclein immunocytochemistry. The inclusions in the cultured cells were identified with HE staining.
  
  
  RESULTS
  The restriction enzyme map suggested that eukaryotic expression vector for human wild-type alpha-synuclein gene was constructed successfully. By EGFP fluorescence, anti-alpha-synuclein immunocytochemistry, it could be observed that the alpha-synuclein protein could aggregate in cytoplasm and the Lewy body-like inclusions found in cytoplasm of cultured cells.
  
  
  CONCLUSIONS
  The over-expression of wild-type alpha-synuclein can induce protein aberrant aggregation and Lewy body-like inclusions formation in cytoplasm of HEK293 cell in vitro.
authors.affiliation: Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R.China.
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Bei-sha
authors.initials: BS
authors.lastname: Tang
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Liao
authors.forename: Xin-xiang
authors.initials: XX
authors.lastname: Yan
authors.forename: Ru-xu
authors.initials: RX
authors.lastname: Zhang
authors.forename: Yu-hu
authors.initials: YH
authors.lastname: Zhang
authors.forename: Jian-guang
authors.initials: JG
authors.lastname: Tang
authors.forename: Li
authors.initials: L
authors.lastname: Cao
authors.forename: Ji-feng
authors.initials: JF
authors.lastname: Guo
authors.forename: Jing
authors.initials: J
authors.lastname: Li
mesh_terms: D002478:Cells, Cultured; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000378:metabolism; D016631:Lewy Bodies / Q000378:metabolism*; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism*; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein