PMID- 25792564 OWN - NLM STAT- MEDLINE DCOM- 20150722 LR - 20220318 IS - 1522-1547 (Electronic) IS - 0193-1857 (Print) IS - 0193-1857 (Linking) VI - 308 IP - 10 DP - 2015 May 15 TI - Glutamine and alanyl-glutamine promote crypt expansion and mTOR signaling in murine enteroids. PG - G831-9 LID - 10.1152/ajpgi.00422.2014 [doi] AB - L-glutamine (Gln) is a key metabolic fuel for intestinal epithelial cell proliferation and survival and may be conditionally essential for gut homeostasis during catabolic states. We show that L-alanyl-L-glutamine (Ala-Gln), a stable Gln dipeptide, protects mice against jejunal crypt depletion in the setting of dietary protein and fat deficiency. Separately, we show that murine crypt cultures (enteroids) derived from the jejunum require Gln or Ala-Gln for maximal expansion. Once expanded, enteroids deprived of Gln display a gradual atrophy of cryptlike domains, with decreased epithelial proliferation, but stable proportions of Paneth and goblet cell differentiation, at 24 h. Replenishment of enteroid medium with Gln selectively activates mammalian target of rapamycin (mTOR) signaling pathways, rescues proliferation, and promotes crypt regeneration. Gln deprivation beyond 48 h leads to destabilization of enteroids but persistence of EGFP-Lgr5-positive intestinal stem cells with the capacity to regenerate enteroids upon Gln rescue. Collectively, these findings indicate that Gln deprivation induces a reversible quiescence of intestinal stem cells and provides new insights into nutritional regulation of intestinal epithelial homeostasis. CI - Copyright (c) 2015 the American Physiological Society. FAU - Moore, Sean R AU - Moore SR AD - Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; sean.moore@cchmc.org. FAU - Guedes, Marjorie M AU - Guedes MM AD - Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil; FAU - Costa, Tie B AU - Costa TB AD - Department of Morphology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil; FAU - Vallance, Jefferson AU - Vallance J AD - Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; FAU - Maier, Elizabeth A AU - Maier EA AD - Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; FAU - Betz, Kristina J AU - Betz KJ AD - Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; FAU - Aihara, Eitaro AU - Aihara E AD - Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, Ohio; and. FAU - Mahe, Maxime M AU - Mahe MM AD - Division of Pediatric Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio. FAU - Lima, Aldo A M AU - Lima AA AD - Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil; FAU - Oria, Reinaldo B AU - Oria RB AD - Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil; FAU - Shroyer, Noah F AU - Shroyer NF AD - Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; LA - eng GR - K02 TW008767/TW/FIC NIH HHS/United States GR - R01 CA142826/CA/NCI NIH HHS/United States GR - P30 DK078392/DK/NIDDK NIH HHS/United States GR - T32 GM063483/GM/NIGMS NIH HHS/United States GR - U19 AI116497/AI/NIAID NIH HHS/United States GR - K02 TW-08767/TW/FIC NIH HHS/United States GR - P30 DK-078392/DK/NIDDK NIH HHS/United States GR - R01 CA-142826/CA/NCI NIH HHS/United States GR - R01 DK-092306/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20150319 PL - United States TA - Am J Physiol Gastrointest Liver Physiol JT - American journal of physiology. Gastrointestinal and liver physiology JID - 100901227 RN - 0 (Dipeptides) RN - 0RH81L854J (Glutamine) RN - EC 2.7.1.1 (mTOR protein, mouse) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - U5JDO2770Z (alanylglutamine) SB - IM MH - Animals MH - Cell Differentiation/physiology MH - Cells, Cultured MH - Dipeptides/*metabolism MH - Epithelial Cells MH - Female MH - Glutamine/*metabolism MH - Intestinal Mucosa/*cytology/*metabolism MH - Male MH - Mice MH - Mice, Inbred C57BL MH - Signal Transduction/physiology MH - Stem Cells/*cytology/*metabolism MH - TOR Serine-Threonine Kinases/*metabolism PMC - PMC4437023 OTO - NOTNLM OT - ERK OT - intestinal organoids OT - l-alanyl-l-glutamine OT - l-glutamine OT - mammalian target of rapamycin EDAT- 2015/03/21 06:00 MHDA- 2015/07/23 06:00 PMCR- 2016/05/15 CRDT- 2015/03/21 06:00 PHST- 2014/11/24 00:00 [received] PHST- 2015/03/13 00:00 [accepted] PHST- 2015/03/21 06:00 [entrez] PHST- 2015/03/21 06:00 [pubmed] PHST- 2015/07/23 06:00 [medline] PHST- 2016/05/15 00:00 [pmc-release] AID - ajpgi.00422.2014 [pii] AID - GI-00422-2014 [pii] AID - 10.1152/ajpgi.00422.2014 [doi] PST - ppublish SO - Am J Physiol Gastrointest Liver Physiol. 2015 May 15;308(10):G831-9. doi: 10.1152/ajpgi.00422.2014. Epub 2015 Mar 19.