Spatiotemporal expression of proprotein convertase subtilisin/ kexin type 5 in the development of anorectal malformations in fetal rats
Spatiotemporal expression of proprotein convertase subtilisin/ kexin type 5 in the development of anorectal malformations in fetal rats
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前蛋白转化酶枯草杆菌蛋白酶/kexin 5型在胎鼠肛门直肠畸形发育中的时空表达
DOI：
10.1007/s10735-017-9736-1
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
白玉作
中科院分区：
文献类型：
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作者：
唐晓冰;陈力;王维林;袁正伟;白玉作
本研究以正常和肛门直肠畸形(ARM)大鼠胚胎为研究对象，研究了Pcsk5在正常大鼠和肛门直肠畸形(ARM)胚胎肛门直肠发育过程中的表达，探讨Pcsk5在ARM发病中的作用。用乙硫脲(GD10)灌胃建立ARM大鼠模型。手术切除GD13~GD16胚胎，用免疫组织化学染色、Western blotting和实时定量RT-PCR检测Pcsk5的时空表达。免疫组织化学染色显示Pcsk5在GD13的泄殖腔上皮(CL)有大量表达。GD14和GD15的直肠中隔和肛膜较薄处可见阳性细胞。而臂胚胎CL上皮仅在GD13~GD15有微弱的Pcsk5表达。Western blotting和Real Time RT-PCR显示Pcsk5在发育中的后肠中的表达呈时间依赖性增加。ARM组Pcsk5表达水平从GD14~GD16均较低(p≤0.05)。这些结果表明，泄殖腔发育到直肠和尿路过程中，Pcsk5的表达下调可能与手臂的形成有关。
This study examined the expression patterns of proprotein convertase subtilisin/kexin type 5 (Pcsk5) during anorectal development in normal and anorectal malfor- mations (ARM) rat embryos, determine the possible role of Pcsk5 in the pathogenesis of ARM. An ARM rat model was developed by the administration of ethylenethiourea gestational day 10 (GD10). Embryos were harvested by surgical excision from GD13 to GD16, and the spatiotem- poral expression of Pcsk5 was evaluated, using immuno- histochemistry staining, Western blotting and real time RT- PCR. Immunohistochemistry staining in normal embryos revealed that Pcsk5 was abundantly expressed on the epithelium of the cloaca (CL) on GD13. On GD14 and GD15, positive cells were noted on the urorectal septum and the thin anal membrane. However, the epithelium of the CL of ARM embryos only faintly expressed Pcsk5 from GD13 to GD15. Western blotting and real time RT-PCR showed time- dependent increase of Pcsk5 expression in the developing hindgut. Pcsk5 expression levels were lower in the ARM group from GD14 to GD16 (p ≤ 0.05). These results indicate that downregulation of Pcsk5 during cloaca development into the rectum and urethra might be related to the forma- tion of ARMs.