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Vol. 16, No. 10, pp. 1234-1246, May 15, 2002
1 Experimental and Molecular Cardiology Group, Academic
Medical Center, University of Amsterdam, Amsterdam, The
Netherlands; 2 Genetically Modified Mice Facility, Academic
Medical Center, University of Amsterdam, Amsterdam, The Netherlands;
3 Division of Molecular Genetics, The Netherlands Cancer
Institute, Amsterdam, The Netherlands; 4 National Research
Council, Center for Muscle Biology and Physiopathology, Department of
Biomedical Sciences, University of Padova, Padova, Italy
During heart development, chamber myocardium forms locally from the
embryonic myocardium of the tubular heart. The atrial natriuretic
factor (ANF) gene is specifically expressed in this developing
chamber myocardium and is one of the first hallmarks of chamber
formation. We investigated the regulatory mechanism underlying this
selective expression. Transgenic analysis shows that a small fragment
of the ANF gene is responsible for the developmental pattern of
endogenous ANF gene expression. Furthermore, this fragment is
able to repress cardiac troponin I (cTnI) promoter
activity selectively in the embryonic myocardium of the
atrioventricular canal (AVC). In vivo inactivation of a T-box factor
(TBE)- or NK2-homeobox factor binding element (NKE) within the
ANF fragment removed the repression in the AVC without
affecting its chamber activity. The T-box family member Tbx2,
encoding a transcriptional repressor, is expressed in the embryonic
myocardium in a pattern mutually exclusive to ANF, thus
suggesting a role in the suppression of ANF. Tbx2 formed a
complex with Nkx2.5 on the ANF TBE-NKE, and was able to
repress ANF promoter activity. Our data provide a potential
mechanism for chamber-restricted gene activity in which the cooperative
action of Tbx2 and Nkx2.5 inhibits expression in the AVC.
[Key Words: Heart development; chamber formation; transgenic mice; ANF; Tbx2; Nkx2.5]
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