title: Abnormal neural crest innervation in Sox10-Venus mice with all-trans retinoic acid-induced anorectal malformations.
abstract: OBJECTIVE
  Despite technical advances in the surgical/medical care of anorectal malformation (ARM), persistent unsatisfactory postoperative bowel habit has been attributed to histopathologic abnormalities of the distal rectum/pouch (DRP) and hypoplasia of anal sphincter muscles (ASM). We used Sox10-Venus mice with ARM induced by all-trans retinoic acid (ATRA) to investigate neural crest cell (NCC) innervation in the DRP and ASM.
  
  
  METHODS
  Pregnant Sox10-Venus mice were administered single doses of 50, 70, or 100 mg/kg of ATRA on embryonic day 8.5 (E8.5) then sacrificed on either E16.5 or E19.5. Bowel specimens comprising the anorectum were examined using fluorescence microscopy without immunohistochemical staining (FMIS). Anti-PGP9.5 was used to delineate ganglion cells and anti-SMA for smooth muscles.
  
  
  RESULTS
  The appropriate dose of ATRA for inducing ARM was 50 mg/kg. Under FMIS, all ARM embryos (n = 5; all high type; 3 male:2 female) had less NCC innervation with thick Venus-positive nerve fibers in the DRP compared with normal embryos (n = 8); there was abnormal NCC innervation in the DRP and absent ASM in ARM mice.
  
  
  CONCLUSIONS
  We are the first to delineate abnormal enteric nervous system innervation in the DRP of ARM mice without using immunohistochemical staining techniques thus allowing specimens to be examined without any distortion.
authors.affiliation: Department of Biochemistry and Biophysics, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, 113-8510, Tokyo, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Suzuki
authors.forename: Katsumi
authors.initials: K
authors.lastname: Miyahara
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
authors.forename: Takashi
authors.initials: T
authors.lastname: Doi
authors.forename: Geoffrey J
authors.initials: GJ
authors.lastname: Lane
authors.forename: Yo
authors.initials: Y
authors.lastname: Mabuchi
authors.forename: Nobuharu
authors.initials: N
authors.lastname: Suzuki
authors.forename: Atsuyuki
authors.initials: A
authors.lastname: Yamataka
authors.forename: Chihiro
authors.initials: C
authors.lastname: Akazawa
mesh_terms: D000015:Abnormalities, Multiple / Q000139:chemically induced* / Q000196:embryology / Q000473:pathology*; D001003:Anal Canal / Q000002:abnormalities* / Q000196:embryology / Q000473:pathology; D000818:Animals; D000071056:Anorectal Malformations; D001006:Anus, Imperforate / Q000139:chemically induced* / Q000196:embryology / Q000473:pathology*; D004195:Disease Models, Animal; D005260:Female; D007422:Intestines / Q000196:embryology / Q000473:pathology*; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence; D009432:Neural Crest / Q000294:innervation* / Q000473:pathology*; D012007:Rectum / Q000002:abnormalities* / Q000196:embryology / Q000473:pathology; D014212:Tretinoin
chemical_list: D014212:Tretinoin