title: A mouse model of pathological small intestinal epithelial cell apoptosis and shedding induced by systemic administration of lipopolysaccharide.
abstract: The gut barrier, composed of a single layer of intestinal epithelial cells (IECs) held together by tight junctions, prevents the entrance of harmful microorganisms, antigens and toxins from the gut lumen into the blood. Small intestinal homeostasis is normally maintained by the rate of shedding of senescent enterocytes from the villus tip exactly matching the rate of generation of new cells in the crypt. However, in various localized and systemic inflammatory conditions, intestinal homeostasis can be disturbed as a result of increased IEC shedding. Such pathological IEC shedding can cause transient gaps to develop in the epithelial barrier and result in increased intestinal permeability. Although pathological IEC shedding has been implicated in the pathogenesis of conditions such as inflammatory bowel disease, our understanding of the underlying mechanisms remains limited. We have therefore developed a murine model to study this phenomenon, because IEC shedding in this species is morphologically analogous to humans. IEC shedding was induced by systemic lipopolysaccharide (LPS) administration in wild-type C57BL/6 mice, and in mice deficient in TNF-receptor 1 (Tnfr1(-/-)), Tnfr2 (Tnfr2(-/-)), nuclear factor kappa B1 (Nfκb1(-/-)) or Nfĸb2 (Nfĸb2(-/-)). Apoptosis and cell shedding was quantified using immunohistochemistry for active caspase-3, and gut-to-circulation permeability was assessed by measuring plasma fluorescence following fluorescein-isothiocyanate-dextran gavage. LPS, at doses ≥0.125 mg/kg body weight, induced rapid villus IEC apoptosis, with peak cell shedding occurring at 1.5 hours after treatment. This coincided with significant villus shortening, fluid exudation into the gut lumen and diarrhea. A significant increase in gut-to-circulation permeability was observed at 5 hours. TNFR1 was essential for LPS-induced IEC apoptosis and shedding, and the fate of the IECs was also dependent on NFκB, with signaling via NFκB1 favoring cell survival and via NFκB2 favoring apoptosis. This model will enable investigation of the importance and regulation of pathological IEC apoptosis and cell shedding in various diseases.
authors.affiliation: Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Williams
authors.forename: Carrie A
authors.initials: CA
authors.lastname: Duckworth
authors.forename: Alastair J M
authors.initials: AJ
authors.lastname: Watson
authors.forename: Mark R
authors.initials: MR
authors.lastname: Frey
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Miguel
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkitt
authors.forename: Robert
authors.initials: R
authors.lastname: Sutton
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Hughes
authors.forename: Lindsay J
authors.initials: LJ
authors.lastname: Hall
authors.forename: Jorge H
authors.initials: JH
authors.lastname: Caamaño
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Campbell
authors.forename: D Mark
authors.initials: DM
authors.lastname: Pritchard
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D004789:Enzyme Activation; D007413:Intestinal Mucosa / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D007421:Intestine, Small / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008070:Lipopolysaccharides / Q000008:administration & dosage / Q000494:pharmacology*; D051379:Mice; D023421:Models, Animal; D016328:NF-kappa B / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D008070:Lipopolysaccharides; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3
references.citation: Anat Rec. 1948 Mar;100(3):357-77
references.pmid: 18906253
references.citation: Sci Signal. 2011 Mar 22;4(165):ra16
references.pmid: 21427409
references.citation: J Endotoxin Res. 2001;7(4):277-80
references.pmid: 11717581
references.citation: Biochem J. 1991 Dec 1;280 ( Pt 2):331-4
references.pmid: 1747105
references.citation: J Exp Med. 1998 Jan 19;187(2):185-96
references.pmid: 9432976
references.citation: J Exp Med. 1999 Jun 7;189(11):1777-82
references.pmid: 10359581
references.citation: Immunity. 2011 May 27;34(5):637-50
references.pmid: 21616434
references.citation: Crit Care Med. 2003 Jun;31(6):1630-7
references.pmid: 12794397
references.citation: J Immunol. 1998 Jan 15;160(2):943-52
references.pmid: 9551933
references.citation: Clin Diagn Lab Immunol. 2005 Jan;12(1):60-7
references.pmid: 15642986
references.citation: Cell. 2001 Feb 23;104(4):487-501
references.pmid: 11239407
references.citation: Nat Rev Immunol. 2010 Feb;10(2):131-44
references.pmid: 20098461
references.citation: Science. 1998 Sep 11;281(5383):1680-3
references.pmid: 9733516
references.citation: J Immunol. 1999 Apr 1;162(7):3749-52
references.pmid: 10201887
references.citation: Immunology. 2004 Oct;113(2):153-62
references.pmid: 15379975
references.citation: Nucleic Acids Res. 2002 May 1;30(9):e36
references.pmid: 11972351
references.citation: Gastroenterology. 2007 Dec;133(6):1769-78
references.pmid: 18054549
references.citation: Lab Invest. 2006 Oct;86(10):1052-63
references.pmid: 16909128
references.citation: J Membr Biol. 1990 Jun;116(2):177-84
references.pmid: 2380981
references.citation: Pharmacol Res. 2002 Aug;46(2):119-27
references.pmid: 12220950
references.citation: J Trauma Acute Care Surg. 2013 Jan;74(1):203-13
references.pmid: 23271096
references.citation: J Biol Chem. 1999 Apr 16;274(16):10689-92
references.pmid: 10196138
references.citation: J Clin Gastroenterol. 2011 Mar;45(3):240-5
references.pmid: 21030873
references.citation: Gut. 2009 Jan;58(1):41-8
references.pmid: 18829978
references.citation: Development. 1990 Dec;110(4):1001-20
references.pmid: 2100251
references.citation: Int J Exp Pathol. 2011 Oct;92(5):366-76
references.pmid: 21838744
references.citation: Gut. 2012 Aug;61(8):1146-53
references.pmid: 22115910
references.citation: Gastroenterology. 2012 Jul;143(1):133-44.e4
references.pmid: 22446197
references.citation: J Exp Med. 2002 Mar 4;195(5):559-70
references.pmid: 11877479
references.citation: Science. 1990 Sep 21;249(4975):1431-3
references.pmid: 1698311
references.citation: Cell Rep. 2012 May 31;1(5):434-43
references.pmid: 22708077
references.citation: Eur J Immunol. 1998 Nov;28(11):3499-505
references.pmid: 9842892
references.citation: Ann Intern Med. 1986 Dec;105(6):883-5
references.pmid: 3777713
references.citation: Nature. 1985 Aug 8-14;316(6028):552-4
references.pmid: 2993897
references.citation: Cell. 2007 Jan 26;128(2):369-81
references.pmid: 17254973
references.citation: Eur J Immunol. 2007 Sep;37(9):2509-17
references.pmid: 17705134
references.citation: J Immunol. 2013 Jan 1;190(1):418-27
references.pmid: 23209324
references.citation: Cell. 1995 Jan 27;80(2):321-30
references.pmid: 7834752
references.citation: J Exp Med. 2011 Aug 29;208(9):1889-900
references.pmid: 21825016
references.citation: Gastroenterology. 2002 Jan;122(1):134-44
references.pmid: 11781288
references.citation: Cytokine. 1993 Jan;5(1):24-30
references.pmid: 8485304
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14635-40
references.pmid: 16192349
references.citation: J Biol Chem. 2011 Jul 1;286(26):22814-24
references.pmid: 21558270
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834