title: Qualitative research of alternatively splice variants of fibronectin in different development stage of mice heart.
abstract: BACKGROUND
  Fibronectin (FN) plays vital roles in cell adhesion, differentiation, proliferation and migration. It is involved in the process of embryonic development and is highly conserved during evolution. The EIIIA and EIIIB of FN show a very high degree of homology among vertebrates. Embryos deleting both EIIIA and EIIIB displayed multiple embryonic cardiovascular defects, implying their crucial role during embryogenesis. The correlation of spliced EIIIB, EIIIA, and IIICS of FN to heart development was studied by observing their chronological expression in mice heart.
  
  
  METHODS
  C57 mice embryos at E11.5, E12.5, E13.5, E14.5, E15.5, E16.5, E17.5, E18.5, E19.5 days, postnatal day 1 (P1d), and adult male mice (3 months) were used. For each alternatively spliced FN1 domain (EIIIB, EIIIA and IIICS), primer pairs were designed for specific amplification. Total RNA was extracted from the heart tissue, reverse transcripted to cDNA, followed by RT-PCR with specific primers. The PCR amplification was verified by agarose gel electrophoresis, showing specific fragments of the expected sizes.
  
  
  RESULTS
  In adult mice heart, only alternatively splice variants of EIIIA-, EIIIB-, IIICS+ were expressed. While in embryonic mice, spliced variant of EIIIA+/-, EIIIB+/-, IIICS+ were observed. The expression of EIIIA and EIIIB changed during heart development.
  
  
  CONCLUSIONS
  FN is crucial for the normal development of the embryonic heart by modulating cardiac neural crest (CNC) proliferation and survival, and maintenance of CNC cells. FN1 gene seems to play a significant role by expression of highly conserved EIIIA and EIIIB in embryonic heart development.
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Lu
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Ma
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Fei-Yu
authors.initials: FY
authors.lastname: Wei
authors.affiliation: 1 Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; 2 Xiamen Diabetes Institute, First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
keywords: EIIIA; EIIIB; Fibronectin (FN); embryonic heart; mice