{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T12:44:20Z","timestamp":1780404260148,"version":"3.54.1"},"reference-count":30,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T00:00:00Z","timestamp":1662422400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["82071685"],"award-info":[{"award-number":["82071685"]}]},{"name":"National Natural Science Foundation of China","award":["81400579"],"award-info":[{"award-number":["81400579"]}]},{"name":"National Natural Science Foundation of China","award":["2019YBKY026"],"award-info":[{"award-number":["2019YBKY026"]}]},{"name":"National Natural Science Foundation of China","award":["2020BCB008"],"award-info":[{"award-number":["2020BCB008"]}]},{"name":"National Natural Science Foundation of China","award":["2020DCD006"],"award-info":[{"award-number":["2020DCD006"]}]},{"name":"National Natural Science Foundation of China","award":["SZSM201812055"],"award-info":[{"award-number":["SZSM201812055"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["82071685"],"award-info":[{"award-number":["82071685"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["81400579"],"award-info":[{"award-number":["81400579"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["2019YBKY026"],"award-info":[{"award-number":["2019YBKY026"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["2020BCB008"],"award-info":[{"award-number":["2020BCB008"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["2020DCD006"],"award-info":[{"award-number":["2020DCD006"]}]},{"name":"Clinical Research Pilot Project of Tongji Hospital","award":["SZSM201812055"],"award-info":[{"award-number":["SZSM201812055"]}]},{"name":"Provincial Key Research and Development Program","award":["82071685"],"award-info":[{"award-number":["82071685"]}]},{"name":"Provincial Key Research and Development Program","award":["81400579"],"award-info":[{"award-number":["81400579"]}]},{"name":"Provincial Key Research and Development Program","award":["2019YBKY026"],"award-info":[{"award-number":["2019YBKY026"]}]},{"name":"Provincial Key Research and Development Program","award":["2020BCB008"],"award-info":[{"award-number":["2020BCB008"]}]},{"name":"Provincial Key Research and Development Program","award":["2020DCD006"],"award-info":[{"award-number":["2020DCD006"]}]},{"name":"Provincial Key Research and Development Program","award":["SZSM201812055"],"award-info":[{"award-number":["SZSM201812055"]}]},{"name":"Science and Technology Innovation Base Platform","award":["82071685"],"award-info":[{"award-number":["82071685"]}]},{"name":"Science and Technology Innovation Base Platform","award":["81400579"],"award-info":[{"award-number":["81400579"]}]},{"name":"Science and Technology Innovation Base Platform","award":["2019YBKY026"],"award-info":[{"award-number":["2019YBKY026"]}]},{"name":"Science and Technology Innovation Base Platform","award":["2020BCB008"],"award-info":[{"award-number":["2020BCB008"]}]},{"name":"Science and Technology Innovation Base Platform","award":["2020DCD006"],"award-info":[{"award-number":["2020DCD006"]}]},{"name":"Science and Technology Innovation Base Platform","award":["SZSM201812055"],"award-info":[{"award-number":["SZSM201812055"]}]},{"name":"Project of Shenzhen San Ming","award":["82071685"],"award-info":[{"award-number":["82071685"]}]},{"name":"Project of Shenzhen San Ming","award":["81400579"],"award-info":[{"award-number":["81400579"]}]},{"name":"Project of Shenzhen San Ming","award":["2019YBKY026"],"award-info":[{"award-number":["2019YBKY026"]}]},{"name":"Project of Shenzhen San Ming","award":["2020BCB008"],"award-info":[{"award-number":["2020BCB008"]}]},{"name":"Project of Shenzhen San Ming","award":["2020DCD006"],"award-info":[{"award-number":["2020DCD006"]}]},{"name":"Project of Shenzhen San Ming","award":["SZSM201812055"],"award-info":[{"award-number":["SZSM201812055"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hirschsprung\u2019s disease (HSCR) is a common developmental anomaly of the gastrointestinal tract in children. The most significant characteristics of aganglionic segments in HSCR are hyperplastic extrinsic nerve fibers and the absence of endogenous ganglion plexus. Double C2 domain alpha (DOC2A) is mainly located in the nucleus and is involved in Ca2+-dependent neurotransmitter release. The loss function of DOC2A influences postsynaptic protein synthesis, dendrite morphology, postsynaptic receptor density and synaptic plasticity. It is still unknown why hyperplastic extrinsic nerve fibers grow into aganglionic segments in HSCR. We detected the expression of DOC2A in HSCR aganglionic segment colons and established three DOC2A-knockdown models in the Neuro-2a cell line, neural spheres and zebrafish separately. First, we detected the protein and mRNA expression of DOC2A and found that DOC2A was negatively correlated with AChE+ grades. Second, in the Neuro-2a cell lines, we found that the amount of neurite outgrowth and mean area per cell were significantly increased, which suggested that the inhibition of DOC2A promotes nerve fiber formation and the neuron\u2019s polarity. In the neural spheres, we found that the DOC2A knockdown was manifested by a more obvious connection of nerve fibers in neural spheres. Then, we knocked down Doc2a in zebrafish and found that the down-regulation of Doc2a accelerates the formation of hyperplastic nerve fibers in aganglionic segments in zebrafish. Finally, we detected the expression of MUNC13-2 (UNC13B), which was obviously up-regulated in Grade3\/4 (lower DOC2A expression) compared with Grade1\/2 (higher DOC2A expression) in the circular muscle layer and longitudinal muscle layer. The expression of UNC13B was up-regulated with the knocking down of DOC2A, and there were protein interactions between DOC2A and UNC13B. The down-regulation of DOC2A may be an important factor leading to hyperplastic nerve fibers in aganglionic segments of HSCR. UNC13B seems to be a downstream molecule to DOC2A, which may participate in the spasm of aganglionic segments of HSCR patient colons.<\/jats:p>","DOI":"10.3390\/ijms231810204","type":"journal-article","created":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T05:17:23Z","timestamp":1662441443000},"page":"10204","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Down-Regulation of Double C2 Domain Alpha Promotes the Formation of Hyperplastic Nerve Fibers in Aganglionic Segments of Hirschsprung\u2019s Disease"],"prefix":"10.3390","volume":"23","author":[{"given":"Jun","family":"Xiao","sequence":"first","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyao","family":"Meng","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luyao","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0015-4371","authenticated-orcid":false,"given":"Yingjian","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou 350001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaosi","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiexiong","family":"Feng","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2669-179X","authenticated-orcid":false,"given":"Zhi","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China"},{"name":"Hubei Clinical Center of Hirschsprung\u2019s Disease and Allied Disorders, Wuhan 430030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1038\/pr.2016.202","article-title":"Hirschsprung\u2019s disease: Clinical dysmorphology, genes, micro-RNAs, and future perspectives","volume":"81","author":"Sergi","year":"2017","journal-title":"Pediatric Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1186\/s13023-020-01362-3","article-title":"ERNICA guidelines for the management of rectosigmoid Hirschsprung\u2019s disease","volume":"15","author":"Kyrklund","year":"2020","journal-title":"Orphanet J. 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