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Spatiotemporal expression of Wnt5a during the development of the striated muscle complex in rats with anorectal
Jie Mi1, Dong Chen1, Xiantan Ren1
1Department of Pediatric Surgery, Shengjing Hospital of China Medical University Liaoning, China.
Unlabelled:
Fecal incontinence and constipation after procedures for anorectal malformations (ARMs) are closely related to the maldevelopment of the striated muscle complex (SMC). Previous studies have demonstrated that myogenic regulatory factors (MRFs) play a significant role in muscle development. Wnt signal pathway is extremely important for MRFs regulation. This study was designed to investigate the spatiotemporal expression pattern of Wnt5a in SMC in ARMs rat embryos.
Materials And Methods:
Anorectal malformation embryos were induced by ethylene thiourea on embryonic day 10 (E10). Expression levels of protein and mRNA of Wnt5a were confirmed by immunohistochemistry staining, western blot and quantitative real-time PCR (qRT-PCR) between normal rat embryos and embryos with ARMs.
Results:
Immunostaining revealed that, on embryonic day 17 (E17), the Wnt5a protein was initially expressed in the SMC in normal embryos. With the growth of pregnancy, the positive staining cells gradually increased. The same time-dependent changes of Wnt5a protein were detected in ARMs embryos. Besides, immunostaining showed that Wnt5a had a significant increase in normal embryos compared with ARMs embryos. Similarly, in Western blot and qRT-PCR, the higher expression of Wnt5a protein and mRNA were remarkable in normal embryos during the SMC development, relatively.
Conclusion:
Our study demonstrated that the downregulation of Wnt5a at the time of SMC development might partly be related to the dysplasia of SMC in ARMs.
Insights
Downregulation of Wnt5a in the striated muscle complex (SMC) may contribute to anorectal malformations (ARMs) in rat embryos. This finding offers insights into the developmental origins of ARM-related functional deficits.
Area of Science:
- Developmental biology
- Molecular genetics
- Anorectal malformations research
Background:
- Fecal incontinence and constipation after anorectal malformations (ARMs) procedures are linked to striated muscle complex (SMC) maldevelopment.
- Myogenic regulatory factors (MRFs) are crucial for muscle development, with Wnt signaling pathways regulating MRFs.
Purpose of the Study:
- To investigate the spatiotemporal expression of Wnt5a in the SMC of rat embryos with ARMs.
- To understand the role of Wnt5a in SMC development in the context of ARMs.
Main Methods:
- Anorectal malformations induced in rat embryos using ethylene thiourea.
- Wnt5a protein and mRNA expression analyzed via immunohistochemistry, Western blot, and qRT-PCR in normal and ARM embryos.
Main Results:
- Wnt5a protein initially expressed in SMC on embryonic day 17 in normal embryos, increasing with development.
- Similar time-dependent Wnt5a expression observed in ARM embryos, but at significantly lower levels compared to normal embryos.
- Western blot and qRT-PCR confirmed higher Wnt5a protein and mRNA expression in normal embryos during SMC development.
Conclusions:
- Downregulation of Wnt5a during SMC development may be a contributing factor to SMC dysplasia in ARMs.
- This study highlights Wnt5a's potential role in the etiology of anorectal malformations.
