Frid M G, Printesva O Y, Chiavegato A, Faggin E, Scatena M, Koteliansky V E, Pauletto P, Glukhova M A, Sartore S
Institute of Experimental Cardiology, Cardiology Research Center, Moscow, Russia.
J Vasc Res. 1993 Sep-Oct;30(5):279-92. doi: 10.1159/000159007.
The myosin heavy-chain (MHC) composition of developing and adult human aortic smooth muscle (SM) was studied by SDS-polyacrylamide gel electrophoresis, Western blotting and indirect immunofluorescence using a panel of anti-MHC antibodies. On 5% SDS gels, three bands of 204, 200 and 196 kDa apparent molecular mass were identified in fetal, infant and adult stages of development. In the extracts from thoracic aorta (upper level), the 204, and 200-kDa bands (designated as SM-1 and SM-2, respectively) were recognized by SM-G4 and SMMS-1 antibodies, raised against a SM antigen, whereas the 196-kDa band was reactive with nonmuscle (NM)-F6 and NM-G2 antiplatelet MHC antibodies. Western blotting and immunofluorescence tests performed on bovine brain and other human NM tissues using NM-F6 and NM-G2 indicated that antigenic targets of the two antibodies resembled that of so-called IIB and IIA NM myosin found in the bovine system, respectively. In the aortic media, SM-1 was expressed throughout development, while SM-2 was upregulated during late fetal and postnatal development. Similarly, the 196-kDa band showed two distinct patterns of immunoreactivity with the anti-NM-MHC antibodies: with NM-G2, antigenicity was equal at all the developmental stages examined, whereas with NM-F6, it diminished during postnatal development. In the upper level, the cellular distribution of NM-G2 and NM-F6 immunoreactivities was similar in the early fetus but quite distinct at later stages of development. In infant and adult subjects, SM cells (SMC) reactive with NM-F6 accumulated predominantly within the intimal layer as well as in some areas of the underlying media as cell foci, whereas NM-G2 homogeneously stained the two layers. In the aorta near the diaphragm (lower level), both antibodies stained the thickened intima but not the underlying media. These data are consistent with the existence of developmental, stage-specific molecular and cellular transitions during vascular SMC maturation in human aortic media. In addition, these data suggest that IIB-like myosin may be expressed in SMC involved specifically in intimal thickening.
采用一组抗肌球蛋白重链（MHC）抗体，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、蛋白质免疫印迹法及间接免疫荧光法，研究了发育中和成人的人类主动脉平滑肌（SM）的MHC组成。在5%的十二烷基硫酸钠凝胶上，在胎儿、婴儿及成人发育阶段鉴定出三条表观分子量分别为204、200和196 kDa的条带。在胸主动脉（较高位置）提取物中，针对平滑肌抗原产生的SM-G4和SMMS-1抗体识别出204 kDa和200 kDa的条带（分别命名为SM-1和SM-2），而196 kDa的条带与非肌肉（NM）-F6和NM-G2抗血小板MHC抗体发生反应。使用NM-F6和NM-G2对牛脑及其他人类非肌肉组织进行蛋白质免疫印迹法和免疫荧光测试表明，这两种抗体的抗原靶点分别类似于在牛系统中发现的所谓IIB和IIA非肌肉肌球蛋白。在主动脉中膜，SM-1在整个发育过程中均有表达，而SM-2在胎儿后期和出生后发育过程中上调。同样，196 kDa的条带与抗非肌肉MHC抗体呈现两种不同的免疫反应模式：与NM-G2反应时，在所检测的所有发育阶段抗原性相同；而与NM-F6反应时，在出生后发育过程中抗原性减弱。在较高位置，NM-G2和NM-F6免疫反应性的细胞分布在胎儿早期相似，但在发育后期则明显不同。在婴儿和成人中，与NM-F6反应的平滑肌细胞（SMC）主要在内膜层以及中膜下层的某些区域以细胞灶的形式聚集，而NM-G2则均匀地染色这两层。在靠近膈肌的主动脉（较低位置），两种抗体均染色增厚的内膜，但不染色其下方的中膜。这些数据与人类主动脉中膜血管平滑肌细胞成熟过程中存在发育阶段特异性的分子和细胞转变相一致。此外，这些数据表明，类似IIB的肌球蛋白可能在特别参与内膜增厚的SMC中表达。
