title: PCSK5 and GDF11 expression in the hindgut region of mouse embryos with anorectal malformations.
abstract: OBJECTIVE
  Retinoid-mediated signal transduction plays a crucial role in the embryonic development of various organs. We previously reported that retinoic acid induced anorectal malformations (ARM) in mice. GDF11 is a TGFβ superfamily molecule and is cleaved and activated by proprotein convertase subtilisin/kexin 5 (PCSK5). PCSK5 (PC5/6) mutations result in an abnormal expression of Hlxb9 and Hox genes, which include known GDF11 targets that are necessary for caudal development in vertebrate embryos. To determine a possible role of the retinoid-mediated signaling pathway in the pathogenesis of ARM, we investigated whether all-trans retinoic acid (ATRA) affected the expression patterns of PCSK5 and GDF11 in ARM-treated mouse embryos.
  
  
  METHODS
  Pregnant ICR-Slc mice were administered 100 mg/kg ATRA by gavage on embryonic day (E) 9.0. Embryos were harvested between days E12 and E18, and mid-sagittal sections of the hindgut region were prepared for immunohistochemistry using antibodies against PCSK5 (PC5/6) and GDF11 (GDF8/11).
  
  
  RESULTS
  Over 95% of the embryos treated with ATRA showed ARM, with rectourethral fistula or rectocloacal fistula, and a short tail. Furthermore, most of these embryos exhibited sacral malformations, tethered spinal cords, and presacral masses resembling those malformations found in caudal regression syndrome. By E14, normal mouse embryos formed a rectum and anus, and the somites behind the hindgut were positive for PC5/6 and GDF8/11. In contrast, in ARM embryos, the somites behind the hindgut were negative for PC5/6 and GDF8/11.
  
  
  CONCLUSIONS
  ATRA treatment affected the caudal development in mouse embryos, resulting in anorectal, sacral, and spinal malformations, and inhibited PCSK5 and GDF11 expression in the hindgut region. These findings indicate that the expression of PCSK5 and GDF11, which plays a crucial role in the organogenesis of the hindgut, was disturbed in the hindgut region when retinoid-mediated signaling was disrupted. This study offers a new insight into the pathogenesis of ARM in mice as affected by the interaction between ATRA and PCSK5/GDF11.
authors.affiliation: Department of Pediatric Surgery, Graduate School of Medical Science, Prefectural University of Medicine, Kyoto, Japan. tom@koto.kpu-m.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Tsuda
authors.forename: N
authors.initials: N
authors.lastname: Iwai
authors.forename: E
authors.initials: E
authors.lastname: Deguchi
authors.forename: O
authors.initials: O
authors.lastname: Kimura
authors.forename: S
authors.initials: S
authors.lastname: Ono
authors.forename: T
authors.initials: T
authors.lastname: Furukawa
authors.forename: Y
authors.initials: Y
authors.lastname: Sasaki
authors.forename: S
authors.initials: S
authors.lastname: Fumino
authors.forename: Y
authors.initials: Y
authors.lastname: Kubota
mesh_terms: D000015:Abnormalities, Multiple / Q000196:embryology / Q000378:metabolism / Q000473:pathology; D000818:Animals; D000071056:Anorectal Malformations; D001006:Anus, Imperforate / Q000196:embryology* / Q000378:metabolism / Q000473:pathology; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D003106:Colon / Q000002:abnormalities / Q000196:embryology* / Q000378:metabolism; D005260:Female; D055412:Growth Differentiation Factors / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D008813:Mice, Inbred ICR; D011247:Pregnancy; D045702:Proprotein Convertase 5 / Q000378:metabolism*; D012003:Rectal Fistula / Q000196:embryology / Q000378:metabolism; D015398:Signal Transduction; D013623:Tail / Q000002:abnormalities / Q000196:embryology; D014212:Tretinoin / Q000008:administration & dosage / Q000009:adverse effects* / Q000378:metabolism
chemical_list: D019485:Bone Morphogenetic Proteins; C119153:Gdf11 protein, mouse; D055412:Growth Differentiation Factors; D014212:Tretinoin; D045702:Proprotein Convertase 5; C473805:propeptide convertase 5, mouse