Cross B E, O'Dea H M, MacPhee D J
Division of Basic Medical Sciences, Health Sciences Centre, Faculty of Medicine, Memorial University of Newfoundland, Rm 5340B, 300 Prince Philip Drive, St John's, NL, Canada A1B 3V6.
Reproduction. 2007 Apr;133(4):807-17. doi: 10.1530/REP-06-0291.
The underlying mechanisms regulating uterine contractions during labour are still poorly understood. Heat shock protein 20 (HSP20) is known to be present at high levels in smooth muscle and implicated in muscle relaxation, but HSP20 expression in the myometrium is completely undetermined. Since HSP20 has been implicated in smooth muscle relaxation, we hypothesized that HSP20 would be highly expressed in the rat myometrium during early and mid-pregnancy when the myometrium is relatively quiescent. Northern blot analysis particularly demonstrated that HSP20 mRNA detection was significantly decreased from day (d) 22 of pregnancy to 1-day post-partum (PP) compared with d6 (P < 0.05). HSP20 mRNA detection was also significantly decreased from d22 to d23 of gestation compared with non-pregnant (NP) samples. Immunoblot analysis showed that detection of HSP20 was significantly decreased at d23 compared with d12 and d15 (P < 0.05). HSP20 detection also significantly decreased at PP compared with d15 (P < 0.05). Immunofluorescence analysis demonstrated that after d15, plasma membrane-associated localization of HSP20 decreased markedly in both circular and longitudinal muscle layers. In addition, HSP20 was detectable near cell membranes at much higher levels in the longitudinal muscle layer of progesterone-treated pregnant rats (delayed labour) at all gestational time points examined, compared with controls. Our results demonstrate that HSP20 mRNA and protein are highly expressed during early and mid-pregnancy and then the expression markedly decreases during late pregnancy and labour. The observed patterns of HSP20 expression are consistent with a potential role for HSP20 in facilitating myometrium quiescence during early and mid-pregnancy.
分娩期间调节子宫收缩的潜在机制仍未得到充分了解。已知热休克蛋白20(HSP20)在平滑肌中高水平存在,并与肌肉松弛有关,但子宫肌层中HSP20的表达完全未明。由于HSP20与平滑肌松弛有关,我们推测HSP20在妊娠早期和中期子宫肌层相对静止时会高表达。Northern印迹分析特别显示,与妊娠第6天相比,从妊娠第22天到产后1天(PP),HSP20 mRNA检测显著降低(P < 0.05)。与非妊娠(NP)样本相比,从妊娠第22天到第23天,HSP20 mRNA检测也显著降低。免疫印迹分析表明,与第12天和第15天相比,第23天HSP20的检测显著降低(P < 0.05)。与第15天相比,产后HSP20检测也显著降低(P < 0.05)。免疫荧光分析表明,在第15天后,HSP20在环行和纵行肌层中的质膜相关定位均明显减少。此外,在所有检查的妊娠时间点,与对照组相比,孕酮处理的妊娠大鼠(分娩延迟)纵行肌层细胞膜附近HSP20的检测水平要高得多。我们的结果表明,HSP20 mRNA和蛋白在妊娠早期和中期高表达,然后在妊娠晚期和分娩期间表达显著降低。观察到的HSP20表达模式与HSP20在妊娠早期和中期促进子宫肌层静止中的潜在作用一致。