title: [Proteomic analysis of the ankle joint bone, ankle joint tissue and spinal cord of clubfoot-like deformity in rat fetuses].
abstract: OBJECTIVE
  To explore the etiology of idiopathic talipes equinovarus (ITEV) in all-trans retinoic acid (ATRA) induced clubfoot-like deformity in rat fetuses with two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS).
  
  
  METHODS
  Clubfoot-like deformity model in rat fetuses was induced with ATRA (135 mg/kg) in gestation day (GD10) pregnant Wistar rats. 2-DE was applied to separate the total proteins of ankle joint tissue, ankle joint bone and spinal cord of the animal models. The Coomassie Brilliant Blue staining gels were analyzed by 2-DE software PDQuest 7.1.0. Selected differential protein spots were identified with peptide mass fingerprinting based on matrix-assisted laser adsorption/ionization time-of-flight mass spectrometry and database searching. xiap, tnnt1 and col2 alpha 1, three genes of the differential proteins, were identified furthermore. Apoptosis study was made in terminal deoxynucleotidyl transferase nick end labeling.
  
  
  RESULTS
  There were many differential expressed proteins in the clubfoot-like deformity model. Out of the differentially expressed proteins,16 protein spots were identified to be differentially expressed in the clubfoot-like deformity model with MS. Three of the 16 protein spots, xiap, tnnt1 and col2 alpha 1 were confirmed to be significantly down-regulated by the RT-PCR, and Xiap was further confirmed to be significantly down-regulated with immunohistochemistry. Another randomly selected gene, ngfr, did not express differently in ATRA-induced clubfoot-like deformity in rat fetuses. The rates of the apoptosis in the spinal, bone of the clubfoot-like deformity fetuses was 5.4 and 10 times of those of the normal fetuses respectively.
  
  
  CONCLUSIONS
  The results suggest that there are certain differently expressed proteins in ankle joint tissue, ankle joint bone and spinal cord of the ATRA-induced clubfoot-like deformity in rat fetuses, and Xiap, sTnT, and Col2 alpha 1 show a significant correlation with ITEV. Ngfr is not correlation with ITEV. Apoptosis plays a key role in the development of ITEV and related to the decreased expression of the Xiap.
authors.affiliation: Department of Medical Genetics, China Medical University, Shenyang, Liaoning, 110001 PR China.
authors.forename: Zeng-gang
authors.initials: ZG
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Ji
authors.forename: Wei-neng
authors.initials: WN
authors.lastname: Fu
authors.forename: Yan-yan
authors.initials: YY
authors.lastname: Zhao
authors.forename: Chun-lian
authors.initials: CL
authors.lastname: Jin
authors.forename: Shi-jun
authors.initials: SJ
authors.lastname: Ji
authors.forename: Kai-lai
authors.initials: KL
authors.lastname: Sun
mesh_terms: D000818:Animals; D000843:Ankle Joint / Q000378:metabolism*; D003025:Clubfoot / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D007150:Immunohistochemistry; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000378:metabolism*; D014212:Tretinoin
chemical_list: D014212:Tretinoin