Early in vertebrate development, mesodermal cells become committed to the cardiogenic lineage. Soon thereafter, these committed precursors begin to differentiate into cardiac myocytes. The molecular basis of cardiac differentiation is not yet understood. Our new data demonstrate, in molecular terms, the embryonic period when committed cardiac progenitor cells are converted to initially differentiated cardiac myocytes. Using an antiserum against the conserved second helix of the myogenic helix-loop-helix (HLH), we provide immnochemical evidence that a HLH protein(s) [called H2] is specifically expressed during the period when committed cardiac progenitors are converted to differentiated myocytes (stage 7-15 of avian development). Western blot and immunoprecipitation analyses demonstrate the presence of a 5OKd protein which is specifically expressed at this time which is reactive with this antibody. Electrophoretic mobility shift assays (EMSA) show the stage specific expression of a DNA binding proteins interactive with muscle specific enhancer sequences and that this DNA/protein complex specifically reacts with the anti-second helix antibody. In a second set of experiments using polymerase chain reaction (PCR) analysis we demonstrate the presence of an additional HLH protein (called Atwist), which is homologous to Drosophila and Xenopus twist and is expressed throughout development.
Our aims i n the proposed grant are clone and characterize these regulatory proteins specifically expressed in the heart using cDNA cloning techniques. Taken together, these studies should provide an understanding of the cellular and molecular basis of cardiac myogenesis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL037675-12
Application #
2430663
Study Section
Cardiovascular and Pulmonary Research A Study Section (CVA)
Project Start
1986-09-30
Project End
1999-05-31
Budget Start
1997-06-01
Budget End
1999-05-31
Support Year
12
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
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