Localization of fos, jun, kit and SCF mRNA in human placenta throughout gestation using in situ RT-PCR.

L Doneda, G Bulfamante, M G Grimoldi, L Volpi, L Larizza
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Abstract

The expression pattern of c-fos, c-jun, c-kit and stem cell factor (SCF) has been investigated in developing human placenta using the highly sensitive technique of in situ reverse transcriptase-polymerase chain reaction (RT-PCR). Specific transcripts of all genes under study were observed in first-trimester placenta sections. c-fos, c-jun, c-kit and SCF transcripts were localized in cells of the villous stroma; fos, jun and kit-specific mRNAs were also found in endothelial cells; fos, kit and SCF mRNAs were detected in villous trophoblast cells. In mid-trimester and term placenta specimens only SCF transcripts were observed, restricted to trophoblast cells. The lack of c-fos transcripts in placenta from the second and third trimesters is a finding that contrasts with data from the literature obtained using extractive techniques. Parallel immunocytochemistry of placenta specimens from the three pregnancy stages under study revealed the fos protein only in first-trimester placenta, in agreement with the in situ RT-PCR findings. We conclude that the in situ RT-PCR technique is most suitable for gene expression studies because of its high level of sensitivity in correctly assigning the signal to specific cell types in complex tissues.

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利用原位RT-PCR技术定位妊娠期人胎盘fos、jun、kit和SCF mRNA。
采用高灵敏的原位逆转录聚合酶链反应(RT-PCR)技术研究了c-fos、c-jun、c-kit和干细胞因子(SCF)在人胎盘发育过程中的表达模式。在妊娠早期胎盘切片中观察到所有基因的特异性转录本。c-fos、c-jun、c-kit和SCF转录本定位于绒毛间质细胞;内皮细胞中也发现了fos、jun和kit特异性mrna;绒毛滋养细胞中检测到fos、kit和SCF mrna。在妊娠中期和足月胎盘标本中,仅观察到滋养层细胞的SCF转录本。妊娠中期和晚期胎盘中缺乏c-fos转录本,这一发现与使用提取技术获得的文献数据形成了对比。平行免疫细胞化学对三个妊娠阶段的胎盘标本进行研究,发现fos蛋白仅在妊娠早期胎盘中存在,与原位RT-PCR结果一致。我们得出结论,原位RT-PCR技术最适合基因表达研究,因为它在将信号正确分配给复杂组织中的特定细胞类型方面具有很高的灵敏度。
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