Altered mRNA and lncRNA expression profiles in the striated muscle complex of anorectal malformation rats
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ORIGINAL ARTICLE
Altered mRNA and lncRNA expression profiles in the striated muscle complex of anorectal malformation rats Zhiya Yao1 · Zhengwei Yuan2 · Yuzuo Bai1 · Hui Gu2 · Huimin Jia1 · Dan Liu2 · Zhonghua Yang1 · Weilin Wang1 Accepted: 1 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Background Striated muscle complex (SMC) dysplasia has been confirmed to contribute to postoperative defecation dysfunction of patients with anorectal malformations (ARMs). To date, the potential molecular mechanisms of SMC dysplasia underlying the development of ARMs have not been clearly explained. This study examined the expression profiles of mRNAs and lncRNAs in the malformed SMC of ARM rats using RNA sequencing (RNA-seq). Methods A rat model of ARMs was established by the intragastric administration of 1% ethylene thiourea (ETU) on an embryonic day 10 (E10). The rats were subjected to euthanasia and the SMC samples were collected on E19. The expression of mRNAs and lncRNAs was analyzed by RNA-seq on the Illumina HiSeq2500 platform. qRT-PCR was used to confirm the results of RNA-seq. Results Compared with the levels in control rats, 1408 mRNAs and 472 lncRNAs were differentially expressed in the SMC of E19 ARM rats. GO and KEGG pathway analyses showed that the top enriched GO terms were mainly related to muscle development and the enriched pathways were associated with muscle and synaptic development. Protein–protein interaction network analysis was also performed using the STRING database. The network map revealed the interaction between the WNT3 protein and NTRK1, NTF4, MUSK, and BMP5 proteins. Finally, the qRT-PCR results further confirmed the RNA-seq data. Conclusion Our findings indicate the involvement of these dysregulated mRNAs and lncRNAs in the pathogenesis of SMC dysplasia in ARMs, providing a theoretical foundation for developing interventions to improve postoperative defecation function. Keywords Striated muscle complex · Anorectal malformations · RNA-seq · mRNA · lncRNA
Introduction Anorectal malformations (ARMs) are the most common malformations of the digestive system in newborns. They occur in about 0.02%–0.06% of neonates [1, 2]. Despite significant improvement in surgical techniques, patients with ARMs may still suffer from postoperative anal dysfunction, * Weilin Wang [email protected] 1
Department of Pediatric Surgery, Shengjing Hospital, China Medical University, 36 Sanhao Street, Shenyang, Liaoning 110004, People’s Republic of China
Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang, Liaoning 110004, People’s Republic of China
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which places a major burden on these patients and their families [3–5]. The presence of striated muscle complex (SMC) dysplasia in ARMs has been demonstrated [6, 7]. SMC plays key roles in defecation control, so the abnormal neural or muscular development of SMC confers the poor functional outcome of ARMs [8]. Therefore, uncovering the molecular mechan
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