Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C

Science

Volumn 276, Issue 5317, 1997, Pages 1404-1407

  Lin, Qing ^a   Schwarz, John ^b   Bucana, Corazon ^c   Olson, Eric N ^a  

^a UNIVERSITY OF TEXAS AT DALLAS   (United States)

^b UNIVERSITY OF TEXAS MEDICAL SCHOOL   (United States)

^c UNIVERSITY OF TEXAS   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALPHA ACTIN; AMINO ACID; ATRIAL NATRIURETIC FACTOR; DNA; MYOSIN HEAVY CHAIN; MYOSIN LIGHT CHAIN; PROTEIN; TRANSCRIPTION FACTOR;

ANIMAL TISSUE; ARTICLE; DNA SEQUENCE; DOWN REGULATION; ELECTROPORATION; EMBRYO; GENE; GENE EXPRESSION; GENE MUTATION; GENOTYPE; HEART; HEART ATRIUM; HEART DEVELOPMENT; HEART VENTRICLE; IN SITU HYBRIDIZATION; MORPHOGENESIS; MOUSE; NONHUMAN; PHENOTYPE; POLYMERASE CHAIN REACTION; PRIORITY JOURNAL; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; SOUTHERN BLOTTING; STEM CELL;

ACYLTRANSFERASES; ANIMALS; GENE EXPRESSION REGULATION, DEVELOPMENTAL; GENE TARGETING; HEART; HEART VENTRICLES; IN SITU HYBRIDIZATION; MICE; MICE, INBRED C57BL; MORPHOGENESIS; MUTAGENESIS; MYOCARDIUM; MYOGENIC REGULATORY FACTORS; STEM CELLS;

EID: 0030911475     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.276.5317.1404     Document Type: Article

References (47)

1. E. N. Olson and D. Srivastava, Science 272, 671 (1996); M. C. Fischman and D. Y. R. Stanier, Circ. Res. 74, 757 (1994).
2. E. N. Olson and D. Srivastava, Science 272, 671 (1996); M. C. Fischman and D. Y. R. Stanier, Circ. Res. 74, 757 (1994).
3. L. A. Gossett, D. J. Kelvin, E. A. Sternberg, E. N. Olson, Mol. Cell. Biol. 9, 5022 (1989).
4. D. G. Edmondson, G. E. Lyons, J. F. Martin, E. N. Olson, Development 120, 1251 (1994).
5. A. E. Chambers et al., Genes Dev. 8, 1324 (1994); M. W. Wong, M. Pisegna, M. F. Lu, D. Leibham, M. Perry, Dev. Biol. 166, 683 (1994); B. Nadal-Ginard and S. V. Subramanian, Mech. Dev. 57, 103 (1996); A. B. Firulli et al., Circ. Res. 78, 196 (1996).
6. A. E. Chambers et al., Genes Dev. 8, 1324 (1994); M. W. Wong, M. Pisegna, M. F. Lu, D. Leibham, M. Perry, Dev. Biol. 166, 683 (1994); B. Nadal-Ginard and S. V. Subramanian, Mech. Dev. 57, 103 (1996); A. B. Firulli et al., Circ. Res. 78, 196 (1996).
7. A. E. Chambers et al., Genes Dev. 8, 1324 (1994); M. W. Wong, M. Pisegna, M. F. Lu, D. Leibham, M. Perry, Dev. Biol. 166, 683 (1994); B. Nadal-Ginard and S. V. Subramanian, Mech. Dev. 57, 103 (1996); A. B. Firulli et al., Circ. Res. 78, 196 (1996).
8. A. E. Chambers et al., Genes Dev. 8, 1324 (1994); M. W. Wong, M. Pisegna, M. F. Lu, D. Leibham, M. Perry, Dev. Biol. 166, 683 (1994); B. Nadal-Ginard and S. V. Subramanian, Mech. Dev. 57, 103 (1996); A. B. Firulli et al., Circ. Res. 78, 196 (1996).
9. E. N. Olson, M. Perry, R. A. Schulz, Dev. Biol. 172, 280 (1995).
10. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
11. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
12. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
13. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
14. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
15. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
16. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991); J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994); J. F. Martin, J. J. Schwarz, E. N. Olson, Proc. Natl. Acad. Sci. U.S.A. 90, 5282 (1993); Y. T. Yu et al., Genes Dev. 9, 1388 (1995); R. E. Breitbart et al., Development 118, 1095 (1993); D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993); J. C. McDermott et al., Mol. Cell. Biol. 13, 2564 (1993).
17. J. D. Molkentin et al., Mol. Cell. Biol. 16, 2627 (1996); Y.-T. Yu, J. Biol. Chem. 271, 24675 (1996).
18. J. D. Molkentin et al., Mol. Cell. Biol. 16, 2627 (1996); Y.-T. Yu, J. Biol. Chem. 271, 24675 (1996).
19. J. Molkentin et al., Mol. Cell. Biol. 16, 3814 (1996).
20. B. Lilly et al., Science 267, 688 (1995); B. A. Bour et al., Genes Dev. 9, 730 (1995); G. Ranganayakulu et al., Dev. Biol. 171, 169 (1995).
21. B. Lilly et al., Science 267, 688 (1995); B. A. Bour et al., Genes Dev. 9, 730 (1995); G. Ranganayakulu et al., Dev. Biol. 171, 169 (1995).
22. B. Lilly et al., Science 267, 688 (1995); B. A. Bour et al., Genes Dev. 9, 730 (1995); G. Ranganayakulu et al., Dev. Biol. 171, 169 (1995).
23. The MEF2C targeting vector was constructed as shown in Fig. 1, with the neomycin resistance gene under control of the phosphoglycerol kinase promoter in the same transcriptional orientation as MEF2C. A thymidine kinase gene under control of the herpes simplex virus promoter pMC1 -HSVtk [S. L. Mansour, K. R. Thomas, M. R. Capecchi, Nature 336, 348 (1988)] was cloned into an Xho I site immediately 5′ of the 5′ arm of genomic homology. All the cloning junctions in the targeting vector were confirmed by DNA sequencing. The targeting vector was linearized by digestion with Not I before etectroporation.
24. The MEF2C targeting vector was introduced by electroporation into 129 ES cells [A. P. McMahon and A. Bradley, Cell 62, 1073 (1990)]. After positive-negative selection, surviving clones were isolated and replica-plated onto SN76/7 fibroblasts in 96-well microtiter plates, and genomic DNA was analyzed on Southern blots.
25. Homologous recombination at the MEF2C locus was detected by Southern blot or PCR analysis of genomic DNA isolated from ES cells, yolk sacs, or tail biopsies. For Southern blots, genomic DNA was digested with Eco RI and hybridized to a probe from a region immediately 3′ of the region of homology used for targeting. For PCR, the following oligonucleotide primers were used: neo primer, 5′-GGCATGCTGGGGATGCGGTGGGCTC-3′; MEF2C MADS box primer, 5′-AGTACAACGAGCCGCACGAGAGCCG-3′; and MEF2C short-arm primer, 5′-GTCACCTTAAGACATAAAGCACCCTCC-3′.
26. R. L. DeHaan, in Organogenesis, R. L. DeHaan and H. Ursprung, Eds. (Holt Rinehart Winston, New York, 1964), pp. 377-419.
27. Heart rates were determined in E9.0 embryos obtained from timed matings of MEF2C heterozygotes. Pregnant females were killed by cervical dislocation; embryos were removed and placed in phosphate-buffered saline at 37°C. Fourteen wild-type and six mutant embryos were analyzed. The average values were statistically significant, as assessed by a standard t test.
28. Q. Lin and E. N. Olson, unpublished data.
29. T. X. O'Brien, K. J. Lee, K. R. Chien, Proc. Natl. Acad. Sci. U.S.A. 90, 5157 (1993).
30. 32P]deoxycytidine triphosphate (dCTP), and gene-specific primers under conditions of linearity for each individual primer set. Most oligonucleotide primers spanned introns. To confirm that samples were not contaminated with genomic DNA, we also performed duplicate PCRs without RT. All PCR products corresponded to the size predicted for the corresponding transcript. PCR cycles were as follows: 99°C for 2 min then 18 to 25 cycles of 96°C for 30 s, 58°C for 30 s, and 72°C for 45 s. PCR products were separated by 6% polyacrylamide gel electrophoresis and quantitated by analysis on a phosphor-Imager (Molecular Dynamics).
31. D. Srivastava, P. Cserjesi, E. N. Olson, Science 270, 1995 (1995); P. Cserjesi, D. Brown, G. E. Lyons, E. N. Olson, Dev. Biol. 170, 664 (1995).
32. D. Srivastava, P. Cserjesi, E. N. Olson, Science 270, 1995 (1995); P. Cserjesi, D. Brown, G. E. Lyons, E. N. Olson, Dev. Biol. 170, 664 (1995).
33. D. Srivastava, T. Thomas, Q. Lin, D. Brown, E. N. Olson, Nature Genetics, in press.
34. C. Biben and R. P. Harvey, personal communication.
35. T. Lints et al., Development 119, 419 (1993).
36. I. Lyons et al., Genes Dev. 9, 1654 (1995); J. Rossant, Circ. Res. 78, 349 (1996).
37. I. Lyons et al., Genes Dev. 9, 1654 (1995); J. Rossant, Circ. Res. 78, 349 (1996).
38. J. D. Molkentin, B. L. Black, J. F. Martin, E. N. Olson, Cell 83, 1125 (1995); S. Kaushal, J. W. Schneider, B. Nadal-Ginard, V. Mahdavi, Science 266, 1236 (1994).
39. J. D. Molkentin, B. L. Black, J. F. Martin, E. N. Olson, Cell 83, 1125 (1995); S. Kaushal, J. W. Schneider, B. Nadal-Ginard, V. Mahdavi, Science 266, 1236 (1994).
40. J. D. Molkentin and B. E. Markham, J. Biol. Chem. 268, 19512 (1993); S. Navankasattusas, H. Zhu, A. V. Garcia, S. M. Evans, K. R. Chien, Mol. Cell. Biol. 12, 1469.
41. J. D. Molkentin and B. E. Markham, J. Biol. Chem. 268, 19512 (1993); S. Navankasattusas, H. Zhu, A. V. Garcia, S. M. Evans, K. R. Chien, Mol. Cell. Biol. 12, 1469.
42. J. Molkentin, Q. Lin, E. N. Olson, unpublished data.
43. G. E. Lyons, B. K. Micales, J. Schwarz, J. F. Martin, E. N. Olson, J. Neurosci. 15, 5727 (1995).
44. J. F. Martin, A. Bradley, E. N. Olson, Genes Dev. 9, 1237 (1995).
45. Sources of probes were as follows: ANF [R. Zeller et al., Genes Dev. 1, 693 (1987); MLC1A [(P. Barton et al., J. Biol. Chem. 263, 12669 (1988)]; MLC2V (16); eHAND and dHAND (18); and Nkx-2.5 (21).
46. Sources of probes were as follows: ANF [R. Zeller et al., Genes Dev. 1, 693 (1987); MLC1A [(P. Barton et al., J. Biol. Chem. 263, 12669 (1988)]; MLC2V (16); eHAND and dHAND (18); and Nkx-2.5 (21).
47. We thank J. Martin for isolating the MEF2C genomic clone, A. Tizenor for assistance with graphics, and D. Srivastava and members of the Olson laboratory for helpful discussions. Supported by grants from NIH, the Muscular Dystrophy Association, the American Heart Association, and the Human Frontiers Sciences Program to E.N.O.