Meer D P, Eddinger T J
Department of Biology, Marquette University, Milwaukee, Wisconsin 53201-1881, USA.
Am J Physiol. 1997 Oct;273(4):C1259-66. doi: 10.1152/ajpcell.1997.273.4.C1259.
The functional significance of the variable expression of the smooth muscle myosin heavy chain (SM-MHC) tail isoforms, SM1 and SM2, was examined at the mRNA level (which correlates with the protein level) in individual permeabilized rabbit arterial smooth muscle cells (SMCs). The length of untethered single permeabilized SMCs was monitored during unloaded shortening in response to increased Ca2+ (pCa 6.0), histamine (1 microM), and phenylephrine (1 microM). Subsequent to contraction, the relative expression of SM1 and SM2 mRNAs from the same individual SMCs was determined by reverse transcription-polymerase chain reaction amplification and densitometric analysis. Correlational analyses between the SM2-to-SM1 ratio and unloaded shortening in saponin- and alpha-toxin-permeabilized SMCs (n = 28) reveal no significant relationship between the SM-MHC tail isoform ratio and unloaded shortening velocity. The best correlations between SM2/SM1 and the contraction characteristics of untethered vascular SMCs were with the minimum length attained following contraction (n = 20 and r = 0.72 for alpha-toxin, n = 8 and r = 0.78 for saponin). These results suggest that the primary effect of variable expression of the SM1 and SM2 SM-MHC tail isoforms is on the cell final length and not on shortening velocity.
在单个透化的兔动脉平滑肌细胞(SMC)中,在mRNA水平(与蛋白质水平相关)检测了平滑肌肌球蛋白重链(SM-MHC)尾部亚型SM1和SM2可变表达的功能意义。在响应Ca2+增加(pCa 6.0)、组胺(1 microM)和去氧肾上腺素(1 microM)时,监测了未束缚的单个透化SMC在无负荷缩短过程中的长度。收缩后,通过逆转录-聚合酶链反应扩增和光密度分析确定来自同一个个别SMC的SM1和SM2 mRNA的相对表达。对皂角苷和α-毒素透化的SMC(n = 28)中SM2与SM1的比率和无负荷缩短之间的相关性分析表明,SM-MHC尾部亚型比率与无负荷缩短速度之间没有显著关系。SM2/SM1与未束缚血管SMC收缩特性之间的最佳相关性是与收缩后达到的最小长度相关(α-毒素处理的n = 20,r = 0.72;皂角苷处理的n = 8,r = 0.78)。这些结果表明,SM1和SM2 SM-MHC尾部亚型可变表达的主要作用是对细胞最终长度,而不是对缩短速度。