Developmental remodeling and shortening of the cardiac outflow tract involves myocyte programmed cell death.
The embryonic outflow tract is a simple tubular structure that connects the single primitive ventricle with the aortic sac and aortic arch arteries. This structure undergoes a complex sequence of morphogenetic processes to become the portion of the heart that aligns the right and left ventricles with the pulmonary artery and aorta. Abnormalities of the outflow tract are involved in many clinically significant congenital cardiac defects; however, the cellular and molecular processes governing the development of this important structure are incompletely understood. Histologic and tissue-tagging studies indicate that the outflow tract tissues compact and are incorporated predominantly into a region of the right ventricle. The hypothesis tested in the current study was that cell death or apoptosis in the muscular portion of the outflow tract is an important cellular mechanism for outflow tract shortening. The tubular outflow tract myocardium was specifically marked by infecting myocytes of the chicken embryo heart with a recombinant replication-defective adenovirus expressing beta-galactosidase (beta-gal) under the control of the cytomegalovirus promoter. Histochemical detection of the beta -gal-labeled outflow tract myocytes revealed that the tubular structure shortened to become a compact ring at the level of the pulmonic infundibulum over several days of development (stages 25-32, embryonic days 4-8). The appearance of apoptotic cardiomyocytes was correlated with OFT shortening by two histologic assays, TUNEL labeling of DNA fragments and AnnexinV binding. The rise and fall in the number of apoptotic myocytes detected by histologic analyses paralleled the change in activity levels of Caspase-3, a protease in the apoptotic cascade, measured in outflow tract homogenates. These results suggest that the elimination of myocytes by programmed cell death is one mechanism by which the outflow tract myocardium remodels to form the proper connection between the ventricular chambers and the appropriate arterial trunks.
Watanabe
M
M
Division of Pediatric Cardiology, Department of Pediatrics, Rainbow Babies and Childrens Hospital, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA. mxw13@po.cwru.edu
Choudhry
A
A
Berlan
M
M
Singal
A
A
Siwik
E
E
Mohr
S
S
Fisher
S A
SA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002642:Chick Embryo; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D005822:Genetic Vectors; D006321:Heart / Q000196:embryology*; D020287:In Situ Nick-End Labeling; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D017304:Annexin A5; D001616:beta-Galactosidase; D053148:Caspase 3; D020169:Caspases