Sandgren J, Scholz TD, Segar JL. ANG II modulation of cardiac growth and remodeling in immature fetal sheep. Am J Physiol Regul Integr Comp Physiol 308: R965-R972, 2015. First published March 25, 2015 doi:10.1152/ajpregu.00034.2015.-ANG II increases fetal blood pressure and stimulates fetal heart growth; however, little is known regarding its direct effects on cardiomyocytes in vivo. We sought to determine whether ANG II stimulates heart growth and cardiomyocyte hypertrophy and/or hyperplasia in utero in the immature fetal heart independent of the effects on cardiac afterload. In twin gestation, fetal sheep at ϳ100 days gestation (term 145 days), one fetus received a chronic (6 days) infusion of ANG II alone (50 g·kg Ϫ1 ·min Ϫ1 ) or ANG II plus nitroprusside (NTP) to attenuate the increase in blood pressure; noninstrumented twins served as controls. ANG II alone, but not ANG II ϩ NTP resulted in a significant increase in heart mass (left and right ventricle ϩ septum, corrected for body weight) compared with controls. ANG II, but not ANG IIϩNTP, also significantly increased cardiomyocyte area compared with control and increased the percentage of binucleated myocytes. ANG II with or without concomitant infusion of NTP increased cardiac PCNA expression, a marker of proliferation. Steady-state protein expression of terminal mitogen-activated protein kinases, cyclin B1, cyclin E1, and p21 were similar among groups. We conclude that in vivo, ANG II increases fetal cardiac mass via cardiomyocyte hypertrophy, differentiation, and to a lesser extent hyperplasia. The effects of ANG II on hypertrophy appear dependent upon the increase in blood pressure (mechanical load), whereas effects on proliferation are load-independent.heart; cardiomyocyte; hypertrophy; hyperplasia STUDIES OF THE FETAL SHEEP heart, long used as a model of human cardiac development, demonstrate marked changes of the myocardium during the last one-third of gestation. In addition to developmental changes in vascularity and myocyte myofibrillar and mitochondrial content, there is a marked increase in the number of cardiomyocytes within the heart, as well as a transition from mononucleated to binucleated (terminally differentiated) myocytes (19,38). In the sheep heart, the transition from mononucleated to binucleated cells begins around 100 days of gestation (term ϳ145 days), such that, at term, ϳ70% of cardiomyocytes are binucleated, or terminally differentiated (19). Both humoral and hemodynamic forces influence cardiac growth and cardiomyocyte proliferation and maturation, although mechanisms governing the normal heart growth process are not well understood (6, 10, 16 -18, 26, 37). Regulation of this developmental process is essential, given that cardiomyocyte endowment is essentially determined by birth and that postnatal proliferative capacity is limited. Disruption of this process, with resultant underendowment of the heart with myocytes, as may occur with fetal hypoxia or undernutrition, may increase susceptibility to cardiac failure later in life...