Novel cell lines promote the discovery of genes involved in early heart development

被引:34
作者
Brunskill, EW
Witte, DP
Yutzey, KE
Potter, SS
机构
[1] Childrens Hosp, Med Ctr, Div Dev Biol, Cincinnati, OH 45229 USA
[2] Childrens Hosp, Med Ctr, Div Pathol, Cincinnati, OH 45229 USA
[3] Childrens Hosp, Med Ctr, Div Mol Cardiovasc Biol, Cincinnati, OH 45229 USA
关键词
cardiac; cell line; Nkx2.5; embryonic; SV40 large T antigen; transgenic mouse; oligonucleotide microarray;
D O I
10.1006/dbio.2001.0313
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Clonal cell lines representing early cardiomyocytes would provide valuable reagents for the dissection of the genetic program of early cardiogenesis. Here we describe the establishment and characterization of cell lines from the hearts of transgenic mice and embryos with SV40 large T antigen expressed in the heart-forming region. Ultrastructure analysis by transmission electron microscopy showed the primitive, precontractile nature of the resulting cells, with the absence of myofilaments, Z lines, and intercalated disks. Immunohistochemistry, RT-PCR, Northern blots, and oligonucleotide microarrays were used to determine the expression levels of thousands of genes in the 1H and ECL-2 cell lines. The resulting gene-expression profiles showed the transcription of early cardiomyocyte genes such as Nkx2.5, GATA4, Tbx5, dHAND, cardiac troponin C, and SM22-alpha. Furthermore, many genes not previously implicated in early cardiac development were expressed. Two of these genes, Hic-5, a possible negative regulator of muscle differentiation, and the transcription enhancing factor TEF-5 were selected and shown by in situ hybridizations to be expressed in the early developing heart. The results show that the 1H and ECL-2 cell lines can be used to discover novel genes expressed in the early cardiomyocyte. (C) 2001 Academic Press
引用
收藏
页码:507 / 520
页数:14
相关论文
共 67 条
[1]  
Alarid ET, 1996, DEVELOPMENT, V122, P3319
[2]  
Ausubel F.M., 1992, CURRENT PROTOCOLS MO
[3]   HEART AND BONE-TUMORS IN TRANSGENIC MICE [J].
BEHRINGER, RR ;
PESCHON, JJ ;
MESSING, A ;
GARTSIDE, CL ;
HAUSCHKA, SD ;
PALMITER, RD ;
BRINSTER, RL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (08) :2648-2652
[4]   Chamber-specific cardiac expression of Tbx5 and heart defects in Holt-Oram syndrome [J].
Bruneau, BG ;
Logan, M ;
Davis, N ;
Levi, T ;
Tabin, CJ ;
Seidman, JG ;
Seidman, CE .
DEVELOPMENTAL BIOLOGY, 1999, 211 (01) :100-108
[5]  
Chapman DL, 1996, DEV DYNAM, V206, P379, DOI 10.1002/(SICI)1097-0177(199608)206:4<379::AID-AJA4>3.0.CO
[6]  
2-F
[7]   TRANSCRIPTIONAL ENHANCER FACTOR-1 DISRUPTION BY A RETROVIRAL GENE TRAP LEADS TO HEART-DEFECTS AND EMBRYONIC LETHALITY IN MICE [J].
CHEN, Z ;
FRIEDRICH, GA ;
SORIANO, P .
GENES & DEVELOPMENT, 1994, 8 (19) :2293-2301
[8]   THE HELA-CELL PROTEIN TEF-1 BINDS SPECIFICALLY AND COOPERATIVELY TO 2 SV40 ENHANCER MOTIFS OF UNRELATED SEQUENCE [J].
DAVIDSON, I ;
XIAO, JH ;
ROSALES, R ;
STAUB, A ;
CHAMBON, P .
CELL, 1988, 54 (07) :931-942
[9]   ABSENCE OF RADIUS AND ULNA IN MICE LACKING HOXA-11 AND HOXD-11 [J].
DAVIS, AP ;
WITTE, DP ;
HSIEHLI, HM ;
POTTER, SS ;
CAPECCHI, MR .
NATURE, 1995, 375 (6534) :791-795
[10]   MORPHOLOGICAL CHARACTERIZATION OF CARDIOMYOCYTES ISOLATED FROM A TRANSPLANTABLE CARDIAC TUMOR DERIVED FROM TRANSGENIC MOUSE ATRIA (AT-1 CELLS) [J].
DELCARPIO, JB ;
LANSON, NA ;
FIELD, LJ ;
CLAYCOMB, WC .
CIRCULATION RESEARCH, 1991, 69 (06) :1591-1600