Loss of MYO5B Leads to Reductions in Na + Absorption With Maintenance of CFTR-Dependent Cl - Secretion in Enterocytes

Gastroenterology. 2018 Dec;155(6):1883-1897.e10. doi: 10.1053/j.gastro.2018.08.025. Epub 2018 Aug 23.

Abstract

Background & aims: Inactivating mutations in MYO5B cause microvillus inclusion disease (MVID), but the physiological cause of the diarrhea associated with this disease is unclear. We investigated whether loss of MYO5B results in aberrant expression of apical enterocyte transporters.

Methods: We studied alterations in apical membrane transporters in MYO5B-knockout mice, as well as mice with tamoxifen-inducible, intestine-specific disruption of Myo5b (VilCreERT2;Myo5bflox/flox mice) or those not given tamoxifen (controls). Intestinal tissues were collected from mice and analyzed by immunostaining, immunoelectron microscopy, or cultured enteroids were derived. Functions of brush border transporters in intestinal mucosa were measured in Ussing chambers. We obtained duodenal biopsy specimens from individuals with MVID and individuals without MVID (controls) and compared transporter distribution by immunocytochemistry.

Results: Compared to intestinal tissues from littermate controls, intestinal tissues from MYO5B-knockout mice had decreased apical localization of SLC9A3 (also called NHE3), SLC5A1 (also called SGLT1), aquaporin (AQP) 7, and sucrase isomaltase, and subapical localization of intestinal alkaline phosphatase and CDC42. However, CFTR was present on apical membranes of enterocytes from MYO5B knockout and control mice. Intestinal biopsies from patients with MVID had subapical localization of NHE3, SGLT1, and AQP7, but maintained apical CFTR. After tamoxifen administration, VilCreERT2;Myo5bflox/flox mice lost apical NHE3, SGLT1, DRA, and AQP7, similar to germline MYO5B knockout mice. Intestinal tissues from VilCreERT2;Myo5bflox/flox mice had increased CFTR in crypts and CFTR localized to the apical membranes of enterocytes. Intestinal mucosa from VilCreERT2;Myo5bflox/flox mice given tamoxifen did not have an intestinal barrier defect, based on Ussing chamber analysis, but did have decreased SGLT1 activity and increased CFTR activity.

Conclusions: Although trafficking of many apical transporters is regulated by MYO5B, trafficking of CFTR is largely independent of MYO5B. Decreased apical localization of NHE3, SGLT1, DRA, and AQP7 might be responsible for dysfunctional water absorption in enterocytes of patients with MVID. Maintenance of apical CFTR might exacerbate water loss by active secretion of chloride into the intestinal lumen.

Keywords: Aquaporin; Mislocalization; Mouse Model; Sodium Hydrogen Exchanger.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Aquaporins / metabolism
  • Chlorides / metabolism*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism*
  • Duodenum / metabolism
  • Duodenum / pathology
  • Enterocytes / metabolism*
  • Gene Silencing
  • Humans
  • Intestinal Mucosa
  • Intestines / cytology
  • Intestines / pathology
  • Malabsorption Syndromes / genetics*
  • Malabsorption Syndromes / pathology
  • Mice
  • Mice, Knockout
  • Microvilli / genetics
  • Microvilli / pathology*
  • Mucolipidoses / genetics*
  • Mucolipidoses / pathology
  • Myosin Type V / genetics*
  • Protein Transport
  • Sodium-Glucose Transporter 1 / metabolism
  • Sodium-Hydrogen Exchanger 3 / metabolism
  • Sodium-Hydrogen Exchangers / metabolism*
  • Sucrase-Isomaltase Complex / metabolism
  • Tamoxifen / administration & dosage

Substances

  • Aqp7 protein, mouse
  • Aquaporins
  • Cftr protein, mouse
  • Chlorides
  • Myo5B protein, mouse
  • Sodium-Glucose Transporter 1
  • Sodium-Hydrogen Exchanger 3
  • Sodium-Hydrogen Exchangers
  • Tamoxifen
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Sucrase-Isomaltase Complex
  • Myosin Type V

Supplementary concepts

  • Microvillus inclusion disease