Surgical method to prevent early death of neonatal rat pups with Hirschsprung disease, thus permitting development of long-term therapeutic approaches

Lincon A Stamp; Enie Lei; Jamie J M Liew; Ruslan V Pustovit; Marlene M Hao; David H Croaker; John B Furness; Cameron D Adams

doi:10.1093/biomethods/bpac004

Surgical method to prevent early death of neonatal rat pups with Hirschsprung disease, thus permitting development of long-term therapeutic approaches

Biol Methods Protoc. 2022 Jan 27;7(1):bpac004. doi: 10.1093/biomethods/bpac004. eCollection 2022.

Authors

Lincon A Stamp^{1

2}, Enie Lei², Jamie J M Liew², Ruslan V Pustovit^{1

2}, Marlene M Hao¹, David H Croaker³, John B Furness^{1

2}, Cameron D Adams²

Affiliations

¹ Department of Anatomy and Physiology, University of Melbourne, Parkville, VIC 3010, Australia.
² Florey Institute of Neuroscience and Mental Health, Parkville, VIC 3010, Australia.
³ Division of Paediatrics and Child Health, Canberra Hospital, Canberra, ACT 2605, Australia.

Abstract

Hirschsprung disease occurs when children are born with no intrinsic nerve cells in varying lengths of the large intestine. In the most severe cases, neurons are also missing from the distal part of the small intestine. Nerve-mediated relaxation of the aganglionic bowel fails and fecal matter accumulates in the more proximal regions of the intestine. This is life threatening. Perforation of the bowel can ensue, causing sepsis and in some cases, death of the infant. Repopulation of the colon with neural stem cells is a potential therapy, but for this to be successful the patient or experimental animal needs to survive long enough for neural precursors to differentiate and make appropriate connections. We have developed a surgical procedure that can be applied to rats with Hirschsprung disease. A stoma was created to allow the normal bowel to empty and a second stoma leading to the aganglionic bowel was also created. This allowed homozygous mutants that would usually die at less than 3 weeks of age to survive into adulthood. During this time, the rats also required post-operative care of their stomas. The interventions we describe provide an animal model of Hirschsprung disease that is suited to assess the effectiveness of cell therapies in the treatment of this condition.

Keywords: Hirschsprung disease; aganglionic colon; enteric nervous system; intestinal surgery; stoma formation.