Yates Dustin T, Clarke Derek S, Macko Antoni R, Anderson Miranda J, Shelton Leslie A, Nearing Marie, Allen Ronald E, Rhoads Robert P, Limesand Sean W
School of Animal and Comparative Biomedical Sciences, The University of Arizona, Tucson, AZ, USA.
Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA, USA.
J Physiol. 2014 Jul 15;592(14):3113-25. doi: 10.1113/jphysiol.2014.272591. Epub 2014 May 23.
Intrauterine growth restriction (IUGR) reduces skeletal muscle mass in fetuses and offspring. Our objective was to determine whether myoblast dysfunction due to intrinsic cellular deficiencies or serum factors reduces myofibre hypertrophy in IUGR fetal sheep. At 134 days, IUGR fetuses weighed 67% less (P < 0.05) than controls and had smaller (P < 0.05) carcasses and semitendinosus myofibre areas. IUGR semitendinosus muscles had similar percentages of pax7-positive nuclei and pax7 mRNA but lower (P < 0.05) percentages of myogenin-positive nuclei (7 ± 2% and 13 ± 2%), less myoD and myogenin mRNA, and fewer (P < 0.05) proliferating myoblasts (PNCA-positive-pax7-positive) than controls (44 ± 2% vs. 52 ± 1%). Primary myoblasts were isolated from hindlimb muscles, and after 3 days in growth media (20% fetal bovine serum, FBS), myoblasts from IUGR fetuses had 34% fewer (P < 0.05) myoD-positive cells than controls and replicated 20% less (P < 0.05) during a 2 h BrdU pulse. IUGR myoblasts also replicated less (P < 0.05) than controls during a BrdU pulse after 3 days in media containing 10% control or IUGR fetal sheep serum (FSS). Both myoblast types replicated less (P < 0.05) with IUGR FSS-supplemented media compared to control FSS-supplemented media. In differentiation-promoting media (2% FBS), IUGR and control myoblasts had similar percentages of myogenin-positive nuclei after 5 days and formed similar-sized myotubes after 7 days. We conclude that intrinsic cellular deficiencies in IUGR myoblasts and factors in IUGR serum diminish myoblast proliferation and myofibre size in IUGR fetuses, but intrinsic myoblast deficiencies do not affect differentiation. Furthermore, the persistent reduction in IUGR myoblast replication shows adaptive deficiencies that explain poor muscle growth in IUGR newborn offspring.
宫内生长受限(IUGR)会降低胎儿及后代的骨骼肌质量。我们的目的是确定由于内在细胞缺陷或血清因子导致的成肌细胞功能障碍是否会降低IUGR胎羊的肌纤维肥大。在134天时,IUGR胎儿体重比对照组轻67%(P<0.05),胴体和半腱肌肌纤维面积更小(P<0.05)。IUGR半腱肌中pax7阳性核和pax7 mRNA的百分比相似,但肌细胞生成素阳性核的百分比更低(P<0.05)(分别为7±2%和13±2%),myoD和肌细胞生成素mRNA更少,增殖的成肌细胞(PNCA阳性-pax7阳性)比对照组少(P<0.05)(44±2%对52±1%)。从后肢肌肉中分离出原代成肌细胞,在生长培养基(20%胎牛血清,FBS)中培养三天后,IUGR胎儿的成肌细胞中myoD阳性细胞比对照组少34%(P<0.05),在2小时的BrdU脉冲期间复制减少20%(P<0.05)。在含有10%对照或IUGR胎羊血清(FSS)的培养基中培养三天后,IUGR成肌细胞在BrdU脉冲期间的复制也比对照组少(P<0.05)。与对照FSS补充培养基相比,两种成肌细胞类型在IUGR FSS补充培养基中的复制都更少(P<0.05)。在促分化培养基(2%FBS)中,IUGR和成肌细胞在5天后肌细胞生成素阳性核的百分比相似,在7天后形成的肌管大小相似。我们得出结论,IUGR成肌细胞的内在细胞缺陷和IUGR血清中的因子会减少IUGR胎儿中成肌细胞的增殖和肌纤维大小,但成肌细胞的内在缺陷不会影响分化。此外,IUGR成肌细胞复制的持续减少显示出适应性缺陷,这解释了IUGR新生后代肌肉生长不良的原因。