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平滑肌肌球蛋白重链亚型在雄性胎儿和新生绵羊中受到发育调控。

Smooth muscle myosin heavy chain isoforms are developmentally regulated in male fetal and neonatal sheep.

作者信息

Chern J, Kamm K E, Rosenfeld C R

机构信息

Department of Pediatrics, University of Texas Southwestern Medical School, Dallas 75235, USA.

出版信息

Pediatr Res. 1995 Nov;38(5):697-703. doi: 10.1203/00006450-199511000-00011.

Abstract

Adult vascular smooth muscle expresses 204-kD (SM1) and 200-kD (SM2) myosin heavy chain (MHC) isoforms. Fetal vascular smooth muscle expresses another 200-kD isoform, MHC-B, that appears to be developmentally regulated. The ontogeny of expression of these MHC isoforms in vascular and nonvascular smooth muscles is not fully understood and may differ. In the present report we examined the ontogeny of these isoforms in aortic and bladder smooth muscle from male fetal (n = 12, 119-140-d gestation; term 145 +/- 5 d) and neonatal (n = 12, 1-33 d) sheep. Tissues were analyzed for total and soluble protein contents. Actin, MHC, and MHC isoforms were analyzed by SDS-PAGE using 3-20% and 4% polyacrylamide gels, respectively. The expression of the adult and fetal 200-kD MHC isoforms were determined by Western analysis. Between 119 d gestation and 33 d neonatal, age-dependent increases (p < 0.02) occurred in bladder actin (16 +/- 0.8 versus 22 +/- 1.4 micrograms/mg of wet weight), MHC (6.5 +/- 0.2 versus 9.7 +/- 1.1) and both soluble (71 +/- 2.9 versus 92 +/- 6.3) and total protein (78 +/- 3.9 versus 103 +/- 5.5). Aortic smooth muscle actin (8.5 +/- 0.7 versus 17 +/- 1.1), MHC (3.1 +/- 0.4 versus 5.2 +/- 0.5), and soluble (44 +/- 2.3 versus 61 +/- 3.0) and total protein (87 +/- 5.8 versus 108 +/- 3.2) also increased (p < 0.01). Aortic SM1 increased (r = 0.79, p < 0.001) during this time, whereas expression of the 200-kD MHC fell (r = -0.79, p < 0.001). In contrast, bladder SM1 fell (r = -0.88, p < 0.001) as the 200-kD MHC rose (r = 0.88, p < 0.001). The type of 200-kD MHC isoform expressed also differed between tissue types; bladder expressed SM2 and little or no MHC-B throughout this phase of development, whereas fetal aorta appeared to express primarily MHC-B, which decreased as adult SM2 expression rose after birth. Expression of smooth muscle proteins and MHC isoforms are developmentally regulated and tissue-dependent, the latter perhaps reflecting developmental differences in organ growth and/or function.

摘要

成年血管平滑肌表达204-kD(SM1)和200-kD(SM2)肌球蛋白重链(MHC)亚型。胎儿血管平滑肌表达另一种200-kD亚型,即MHC-B,其表达似乎受发育调控。这些MHC亚型在血管和非血管平滑肌中的表达个体发生情况尚未完全了解,可能存在差异。在本报告中,我们研究了雄性胎儿(n = 12,妊娠119 - 140天;足月为145±5天)和新生(n = 12,1 - 33天)绵羊的主动脉和膀胱平滑肌中这些亚型的个体发生情况。分析组织的总蛋白和可溶性蛋白含量。分别使用3 - 20%和4%的聚丙烯酰胺凝胶通过SDS-PAGE分析肌动蛋白、MHC和MHC亚型。通过蛋白质印迹分析确定成年和胎儿200-kD MHC亚型的表达。在妊娠119天至出生后33天期间,膀胱肌动蛋白(16±0.8对22±1.4微克/毫克湿重)、MHC(6.5±0.2对9.7±1.1)以及可溶性蛋白(71±2.9对92±6.3)和总蛋白(78±3.9对103±5.5)均随年龄增长而增加(p < 0.02)。主动脉平滑肌肌动蛋白(8.5±0.7对17±1.1)、MHC(3.1±0.4对5.2±0.5)以及可溶性蛋白(44±2.3对61±3.0)和总蛋白(87±5.8对108±3.2)也增加(p < 0.01)。在此期间,主动脉SM1增加(r = 0.79,p < 0.001),而200-kD MHC的表达下降(r = -0.79,p < 0.001)。相反,膀胱SM1下降(r = -0.88,p < 0.001),而200-kD MHC上升(r = 0.88,p < 0.001)。不同组织类型中表达的200-kD MHC亚型类型也有所不同;在整个发育阶段,膀胱表达SM2且几乎不表达或不表达MHC-B,而胎儿主动脉似乎主要表达MHC-B,出生后随着成年SM2表达上升,MHC-B表达下降。平滑肌蛋白和MHC亚型的表达受发育调控且具有组织依赖性,后者可能反映了器官生长和/或功能的发育差异。

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