Nutritional stimulation by in-ovo feeding modulates cellular proliferation and differentiation in the small intestinal epithelium of chicks

• Published in: Animal Nutrition Vol. 8; no. 1; pp. 91 - 101
• Main Authors: Reicher, Naama, Melkman-Zehavi, Tal, Dayan, Jonathan, Wong, Eric A., Uni, Zehava
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.03.2022; KeAi Publishing; KeAi Communications Co., Ltd
• Subjects: animal nutrition; antigens; cell proliferation; Chick; chicks; Differentiation; epithelial cells; fluorescent antibody technique; G-protein coupled receptors; glutamine; in situ hybridization; In-ovo feeding; Intestinal development; intestinal mucosa; leucine; Multipotent cell; Original; peptide transporters; Proliferation; small intestine; stem cells; surface area; transcription factors; villi; In-ovo feeding; Proliferation; Differentiation; Chick; Intestinal development; Multipotent cell; Multipotent cells
• DOI: 10.1016/j.aninu.2021.06.010
• ISSN: 2405-6545; 2405-6383

Nutritional stimulation of the developing small intestine of chick embryos can be conducted by in-ovo feeding (IOF). We hypothesized that IOF of glutamine and leucine can enhance small intestinal development by promoting proliferation and differentiation of multipotent small intestinal epithelial cells. Broiler embryos (n = 128) were subject to IOF of glutamine (IOF-Gln), leucine (IOF-Leu), NaCl (IOF-NaCl) or no injection (control) at embryonic d 17 (E 17). Multipotent, progenitor and differentiated cells were located and quantified in the small intestinal epithelium between E 17 and d 7 after hatch (D 7) in all treatment groups by immunofluorescence of SRY-box transcription factor 9 (Sox9) and proliferating cell nuclear antigen (PCNA), in-situ hybridization of leucine-rich repeat containing G-protein coupled receptor 5 (Lgr5) and peptide transporter 1 (PepT1) and histochemical goblet cell staining. The effects of IOF treatments at E 19 (48 h post-IOF), in comparison to control embryos, were as follows: total cell counts increased by 40%, 33% and 19%, and multipotent cell counts increased by 52%, 50% and 38%, in IOF-Gln, IOF-Leu and IOF-NaCl embryos, respectively. Only IOF-Gln embryos exhibited a significance, 36% increase in progenitor cell counts. All IOF treatments shifted Lgr5+ stem cell localizations to villus bottoms. The differentiated, PepT1+ region of the villi was 1.9 and 1.3-fold longer in IOF-Gln and IOF-Leu embryos, respectively, while goblet cell densities decreased by 20% in IOF-Gln embryos. Post–hatch, crypt and villi epithelial cell counts were significantly higher IOF-Gln chicks, compared to control chicks (P < 0.05). We conclude IOF of glutamine stimulates small intestinal maturation and functionality during the peri-hatch period by promoting multipotent cell proliferation and differentiation, resulting in enhanced compartmentalization of multipotent and differentiated cell niches and expansions of the absorptive surface area.