Modeling Age-Dependent Developmental Changes in the Expression of Genes Involved in Citrulline Synthesis Using Pig Enteroids

Authors

Mahmoud A Mohammad
Inka C Didelija
Barbara Stoll
Douglas G Burrin
Juan C Marini

Publication Date

11-1-2020

Journal

Physiological Reports

DOI

10.14814/phy2.14565

PMID

33181004

PMCID

PMC7660678

PubMedCentral® Posted Date

11-12-2020

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Animals, Animals, Newborn, Citrulline, Duodenum, Female, Gene Expression Regulation, Developmental, Intestinal Mucosa, Jejunum, Male, Swine

Abstract

metabolism of citrulline and arginine are well characterized. Enteroids, a novel ex-vivo model that recreates the three-dimensional structure of the intestinal crypt-villus unit, have shown to replicate molecular and physiological profiles of the intestinal segment from where they originated ("location memory").

Objective: The present study tested the hypothesis that enteroids recapitulate the developmental changes observed in vivo regarding citrulline production in pigs ("developmental memory").

Methods: Preterm (10- and 5-d preterm) and term pigs at birth, together with 7- and 35-d-old pigs were studied. Gene expression was measured in jejunal samples and in enteroids derived from this segment. Whole body citrulline production was measured by isotope dilution and enteroid citrulline production by accumulation in the media.

Results: With the exception of arginase I and inducible nitric oxide synthase, all the genes investigated expressed in jejunum were expressed by enteroids. In the jejunum, established markers of development (lactase and sucrase-isomaltase), as well as genes that code for enzymes involved in the production and utilization of citrulline and arginine, underwent the ontogenic changes described in the literature. However, enteroid expression of these genes, as well as citrulline production, failed to recapitulate the changes observed in vivo.

Conclusions: Under culture conditions used in our study, enteroids derived from jejunal crypts of pigs at different ages failed to replicate the gene expression observed in whole tissue and whole body citrulline production. Additional extracellular cues may be needed to reproduce the age-dependent phenotype.