Top

Published in:

01-12-2023 | Megacolon | Original Article

A novel mouse model of intestinal neuronal dysplasia: visualization of the enteric nervous system

Authors: Naho Fujiwara, Katsumi Miyahara, Dorothy Lee, Nana Nakazawa-Tanaka, Chihiro Akazawa, Masahiko Hatano, Agostino Pierro, Atsuyuki Yamataka

Published in: Pediatric Surgery International | Issue 1/2023

Abstract

Purpose

Intestinal neuronal dysplasia (IND) is a congenital anomaly affecting gastrointestinal neural innervation, but the pathogenesis remains unclear. The homozygous Ncx/Hox11L.1 knockout (Ncx^−/−) mice exhibit megacolon and enteric ganglia anomalies, resembling IND phenotypes. Sox10-Venus transgenic mouse were used to visualize enteric neural crest cells in real time. This study aims to establish a novel mouse model of Sox10-Venus⁺/Ncx^−/− mouse to study the pathogenesis of IND.

Methods

Sox10-Venus⁺/Ncx^−/− (Ncx^−/−) (n = 8) mice and Sox10-Venus⁺/Ncx^+/+ controls (control) (n = 8) were euthanized at 4–5 weeks old, and excised intestines were examined with fluorescence microscopy. Immunohistochemistry was performed on tissue sections with neural marker Tuj1.

Results

Ncx^−/− mice exhibited dilated cecum and small intestine. Body weight of Ncx^−/− mice was lower with higher ratio of small intestine length relative to body weight. The neural network (Sox10-Venus) was observed along the intestine wall in Ncx^−/− and control mice without staining. Ectopic and increased expression of Tuj1 was observed in both small intestine and proximal colon of Ncx^−/− mice.

Conclusion

This study has established a reliable animal model that exhibits characteristics similar to patients with IND. This novel mouse model can allow the easy visualization of ENS in a time- and cost-effective way to study the pathogenesis of IND.

Kapur RP, Reyes-Mugica M (2019) Intestinal neuronal dysplasia type B: an updated review of a problematic diagnosis. Arch Pathol Lab Med 143:235–243. https://doi.org/10.5858/arpa.2017-0524-RACrossRefPubMed

Yamataka A, Hatano M, Kobayashi H, Wang K, Miyahara K, Sueyoshi N et al (2001) Intestinal neuronal dysplasia-like pathology in Ncx/Hox11L.1 gene-deficient mice. J Pediatr Surg 36:1293–1296. https://doi.org/10.1053/jpsu.2001.25797CrossRefPubMed

Meier-Ruge W (1971) Casuistic of colon disorder with symptoms of Hirschsprung’s disease (author’s transl). Verh Dtsch Ges Pathol 55:506–510PubMed

Liu W, Zhou T, Tian J, Yu X, Ren C, Cao Z et al (2022) Role of GDNF, GFRα1 and GFAP in a bifidobacterium-intervention induced mouse model of intestinal neuronal dysplasia. Front Pediatr. https://doi.org/10.3389/fped.2021.795678CrossRefPubMedPubMedCentral

Wang D, Gao N, Zhou T, Zhang Q, Wang J, Li A (2020) Effect of Neuroligin1 and Neurexin1 on the colonic motility in a mouse model of neuronal intestinal dysplasia. Gastroenterol Res Pract 2020:1–9. https://doi.org/10.1155/2020/9818652CrossRef

Hatano M, Aoki T, Dezawa M, Yusa S, Iitsuka Y, Koseki H et al (1997) A novel pathogenesis of megacolon in Ncx/Hox11L.1 deficient mice. J Clin Investig 100:795–801. https://doi.org/10.1172/JCI119593CrossRefPubMedPubMedCentral

Shirasawa S, Yunker AMR, Roth KA, Brown GA, Horning S, Korsmeyer SJ (1997) Enx (Hox11L1)-deficient mice develop myenteric neuronal hyperplasia and megacolon. Nat Med 3:646–650. https://doi.org/10.1038/nm0697-646CrossRefPubMed

Wen X-Y, Tang S, Breitman ML (1994) Genetic mapping of two mouse homeobox genes Tlx-1 and Tlx-2 to murine chromosomes 19 and 6. Genomics 24:388–390. https://doi.org/10.1006/geno.1994.1634CrossRefPubMed

Dear TN, Sanchez-Garcia I, Rabbitts TH (1993) The HOX11 gene encodes a DNA-binding nuclear transcription factor belonging to a distinct family of homeobox genes. Proc Natl Acad Sci 90:4431–4435. https://doi.org/10.1073/pnas.90.10.4431CrossRefPubMedPubMedCentral

10.

Hatano M, Iitsuka Y, Yamamoto H, Dezawa M, Yusa S, Kohno Y et al (1997) Ncx, a Hox11 related gene, is expressed in a variety of tissues derived from neural crest cells. Anat Embryol (Berl) 195:419–425. https://doi.org/10.1007/s004290050061CrossRefPubMed

11.

Kelsh RN (2006) Sorting outSox10 functions in neural crest development. BioEssays 28:788–798. https://doi.org/10.1002/bies.20445CrossRefPubMed

12.

Bondurand N, Natarajan D, Barlow A, Thapar N, Pachnis V (2006) Maintenance of mammalian enteric nervous system progenitors by SOX10 and endothelin 3 signalling. Development 133:2075–2086. https://doi.org/10.1242/dev.02375CrossRefPubMed

13.

Bondurand N, Natarajan D, Thapar N, Atkins C, Pachnis V (2003) Neuron and glia generating progenitors of the mammalian enteric nervous system isolated from foetal and postnatal gut cultures. Development 130:6387–6400. https://doi.org/10.1242/dev.00857CrossRefPubMed

14.

Shibata S, Yasuda A, Renault-Mihara F, Suyama S, Katoh H, Inoue T et al (2010) Sox10- Venus mice: a new tool for real-time labeling of neural crest lineage cells and oligodendrocytes. Mol Brain 3:31. https://doi.org/10.1186/1756-6606-3-31CrossRefPubMedPubMedCentral

15.

Miyahara K, Kato Y, Koga H, Lane GJ, Inoue T, Akazawa C et al (2010) Abnormal enteric innervation identified without histopathologic staining in aganglionic colorectum from a mouse model of Hirschsprung’s disease. J Pediatr Surg 45:2403–2407. https://doi.org/10.1016/j.jpedsurg.2010.08.039CrossRefPubMed

16.

Fujiwara N, Miyahara K, Nakazawa-Tanaka N, Akazawa C, Yamataka A (2022) In vitro investigation of the differentiation of enteric neural crest-derived cells following transplantation of aganglionic gut in a mouse model. Pediatr Surg Int 38:755–759. https://doi.org/10.1007/s00383-022-05105-2CrossRefPubMed

17.

Granero Cendón R, Millán López A, Moya Jiménez MJ, López Alonso M (2007) De Agustín Asensio JC [Intestinal neuronal dysplasia: association with digestive malformations]. Cir Pediatr 20:166–168PubMed

18.

Puri P, Rolle U (2004) Variant Hirschsprung’s disease. Semin Pediatr Surg 13:293–299. https://doi.org/10.1053/j.sempedsurg.2004.10.017CrossRefPubMed

19.

Puri P, Gosemann J-H (2012) Variants of Hirschsprung disease. Semin Pediatr Surg 21:310–318. https://doi.org/10.1053/j.sempedsurg.2012.07.005CrossRefPubMed

20.

Fadda B, Maier W, Meier-Ruge W, Schärli A, Daum R (1983) Neuronale intestinale DysplasieEine kritische 10-Jahres-Analyse klinischer und bioptischer Diagnostik. Eur J Pediatr Surg 38:305–311. https://doi.org/10.1055/s-2008-1059994CrossRef

21.

Puri P (1997) Variant Hirschsprung’s disease. J Pediatr Surg 32:149–157. https://doi.org/10.1016/S0022-3468(97)90170-6CrossRefPubMed

22.

Gath R, Goessling A, Keller K-M, Koletzko S, Coerdt W, Müntefering H et al (2001) Analysis of the RET, GDNF, EDN3, and EDNRB genes in patients with intestinal neuronal dysplasia and Hirschsprung disease. Gut 48:671–675. https://doi.org/10.1136/gut.48.5.671CrossRefPubMedPubMedCentral

23.

Sánchez-Mejías A, Fernández RM, Antiñolo G, Borrego S (2010) A new experimental approach is required in the molecular analysis of intestinal neuronal dysplasia type B patients. Exp Ther Med 1:999–1003. https://doi.org/10.3892/etm.2010.140CrossRefPubMedPubMedCentral

24.

Borghini S, Di DM, Prato AP, Lerone M, Martucciello G, Jasonni V et al (2009) Search for pathogenetic variants of the SPRY2 gene in intestinal innervation defects. Intern Med J 39:335–337. https://doi.org/10.1111/j.1445-5994.2009.01907.xCrossRefPubMed

25.

Fava M, Borghini S, Cinti R, Cusano R, Seri M, Lerone M et al (2002) HOX11L1: a promoter study to evaluate possible expression defects in intestinal motility disorders. Int J Mol Med 10:101–106PubMed

26.

Costa M (2000) Evaluation of the HOX11L1 gene as a candidate for congenital disorders of intestinal innervation. J Med Genet 37:9e–99. https://doi.org/10.1136/jmg.37.7.e9CrossRef

27.

Sacher P, Briner J, Hanimann B (1993) Is neuronal intestinal dysplasia (NID) a primary disease or a secondary phenomenon? Eur J Pediatr Surg 3:228–230. https://doi.org/10.1055/s-2008-1063549CrossRefPubMed

28.

von Boyen GBT (2002) Abnormalities of the enteric nervous system in heterozygous endothelin B receptor deficient (spotting lethal) rats resembling intestinal neuronal dysplasia. Gut 51:414–419. https://doi.org/10.1136/gut.51.3.414CrossRef

29.

Taketomi T, Yoshiga D, Taniguchi K, Kobayashi T, Nonami A, Kato R et al (2005) Loss of mammalian Sprouty2 leads to enteric neuronal hyperplasia and esophageal achalasia. Nat Neurosci 8:855–857. https://doi.org/10.1038/nn1485CrossRefPubMed

30.

Yanai T, Kobayashi H, Yamataka A, Lane GJ, Miyano T, Hayakawa T et al (2004) Acetylcholine-related bowel dysmotility in homozygous mutant NCX/HOX11L.1-deficient (NCX^−/−) mice—evidence that acetylcholine is implicated in causing intestinal neuronal dysplasia. J Pediatr Surg 39:927–930. https://doi.org/10.1016/j.jpedsurg.2004.02.004CrossRefPubMed

31.

Young HM, Bergner AJ, Müller T (2003) Acquisition of neuronal and glial markers by neural crest-derived cells in the mouse intestine. J Comp Neurol 456:1–11. https://doi.org/10.1002/cne.10448CrossRefPubMed

32.

Liu Y-R, Ba F, Cheng L-J, Li X, Zhang S-W, Zhang S-C (2019) Efficacy of Sox10 promoter methylation in the diagnosis of intestinal neuronal dysplasia from the peripheral blood. Clin Transl Gastroenterol 10:e00093. https://doi.org/10.14309/ctg.0000000000000093CrossRefPubMedPubMedCentral

Title: A novel mouse model of intestinal neuronal dysplasia: visualization of the enteric nervous system
Authors: Naho Fujiwara
Katsumi Miyahara
Dorothy Lee
Nana Nakazawa-Tanaka
Chihiro Akazawa
Masahiko Hatano
Agostino Pierro
Atsuyuki Yamataka
Publication date: 01-12-2023
Publisher: Springer Berlin Heidelberg
Keyword: Megacolon
Published in: Pediatric Surgery International / Issue 1/2023
Print ISSN: 0179-0358
Electronic ISSN: 1437-9813
DOI: https://doi.org/10.1007/s00383-023-05585-w

Keynote webinar | Spotlight on medication adherence

Springer Medicine

A novel mouse model of intestinal neuronal dysplasia: visualization of the enteric nervous system

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2023

Risk factors and outcomes in pediatric blunt cardiac injuries

Correction: Diagnostic performance of serum pentraxin-3 in pediatric acute appendicitis: a prospective diagnostic validation study

Clinical experience of thoracoplasty with absorbable rib substitutes

Correction to: Surgical management strategy of alimentary tract duplication involving the rectum in children

Non-operative management of acute appendicitis in children

Surgical management strategy of alimentary tract duplication involving the rectum in children