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单细胞水平上平滑肌肌球蛋白重链表达的异质性。

Heterogeneity of smooth muscle myosin heavy chain expression at the single cell level.

作者信息

Meer D P, Eddinger T J

机构信息

Department of Biology, Marquette University, Milwaukee, Wisconsin 53233, USA.

出版信息

Am J Physiol. 1996 Jun;270(6 Pt 1):C1819-24. doi: 10.1152/ajpcell.1996.270.6.C1819.

Abstract

Expression of the smooth muscle myosin heavy chain (SM-MHC) isoforms was examined in individual rabbit arterial smooth muscle cells with the use of reverse transcription-polymerase chain reactions (RT-PCR). The RT-PCR amplification protocol used oligonucleotide primers complementary to regions that flank the alternative exon that encodes the nine unique amino acids found in the carboxy terminal domain of SM2. RT-PCR products of SM1 and SM2 mRNA differ in length and electrophoretic mobility. Partial DNA sequencing of the PCR products confirmed SM1 and SM2 identity. Densitometric analyses of adjacent samples extracted from the same tissues and processed for SM2-to-SM1 protein and PCR-amplified SM2-to-SM1 RNA ratios exhibited a high correlation (R = 0.92). RT-PCR-amplified SM2-to-SM1 mRNA ratios of individual adult rabbit arterial cells ranged from 0.0 to 1.8 (n = 59), whereas multicellular vascular samples varied much less (0.4-0.6, n = 5). These results indicate that individual cells within a blood vessel differ significantly in SM-MHC expression. This difference may be important for the regulation of contraction in these vessels.

摘要

利用逆转录-聚合酶链反应(RT-PCR)检测了兔动脉平滑肌单个细胞中平滑肌肌球蛋白重链(SM-MHC)亚型的表达情况。RT-PCR扩增方案使用了与编码SM2羧基末端结构域中九个独特氨基酸的可变外显子侧翼区域互补的寡核苷酸引物。SM1和SM2 mRNA的RT-PCR产物在长度和电泳迁移率上有所不同。PCR产物的部分DNA测序证实了SM1和SM2的一致性。对从同一组织中提取并用于检测SM2与SM1蛋白及PCR扩增的SM2与SM1 RNA比值的相邻样本进行光密度分析,结果显示高度相关(R = 0.92)。成年兔动脉单个细胞的RT-PCR扩增SM2与SM1 mRNA比值范围为0.0至1.8(n = 59),而多细胞血管样本的变化则小得多(0.4 - 0.6,n = 5)。这些结果表明,血管内的单个细胞在SM-MHC表达上存在显著差异。这种差异可能对这些血管收缩的调节很重要。

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