Bacterial colonization leads to the colonic secretion of RELMβ/FIZZ2, a novel goblet cell-specific protein

doi:10.1016/j.gastro.2003.07.009

Gastroenterology

Volume 125, Issue 5, November 2003, Pages 1388-1397

https://doi.org/10.1016/j.gastro.2003.07.009 Get rights and content

Abstract

Background & Aims:

Goblet cells are highly polarized exocrine cells found throughout the small and large intestine that have a characteristic morphology due to the accumulation of apical secretory granules. These granules contain proteins that play important physiologic roles in cellular protection, barrier function, and proliferation. A limited number of intestinal goblet cell-specific proteins have been identified. In this study, we investigate the expression and regulation of RELMβ, a novel colon-specific gene.

Methods:

The regulation of RELMβ messenger RNA expression was determined in LS174T, Caco-2, and HT-29 cell lines in response to stimulation with interleukin 13 and lipopolysaccharide. Quantitative reverse-transcription polymerase chain reaction, immunoblots, and immunohistochemistry were used to examine the expression of RELMβ in BALB/c and C.B17.SCID mice housed in conventional, germ-free, and gnotobiotic environments.

Results:

Messenger RNA for RELMβ is restricted to the undifferentiated, proliferating colonic epithelium. Immunohistochemistry shows that this protein is expressed in goblet cells located primarily in the distal half of the colon and cecum with lower levels detectable in the proximal colon. High levels of RELMβ can be detected in the stool of mice and humans, where it exists as a homodimer under nonreducing conditions. Interestingly, the secretion of RELMβ is dramatically reduced in germ-free mice. Furthermore, introduction of germ-free mice into a conventional environment results in enhanced expression and robust secretion of RELMβ within 48 hours.

Conclusions:

These studies define a new goblet cell-specific protein and provide the first evidence that colon-specific gene expression can be regulated by colonization with normal enteric bacteria.

Section snippets

Animal studies

Germ-free BALB/c and C.B17.SCID mice were bred and housed in the germ-free facility of the Department of Biology at the University of Pennsylvania (Philadelphia, PA) under sterile conditions in Trexler isolators (Standard Safety, McHenry, IL). Conventionally reared BALB/c mice were purchased from Jackson Laboratories (Bar Harbor, ME) and housed in the animal facility of the Department of Biology at the University of Pennsylvania. To compare RELMβ expression under germ-free and conventional

RELMβ is expressed in goblet cells

Immunolocalization of RELMβ protein in the mouse colon was performed using a polyclonal, affinity-purified antibody to the carboxyl terminus. RELMβ is expressed throughout the colon, with the highest level of expression observed in the distal half of the colon and the cecum (Figure 1, Figure 2). This is consistent with the pattern of mRNA expression.15 Blocking experiments using the peptide immunogen show that the detection of RELMβ using this antibody is highly specific (Figure 1B). The

Discussion

The intestinal epithelium is a mucosal surface at the interface between luminal contents of the alimentary tract and the internal environment of the animal. More than simply a barrier, the intestinal epithelium is an active interface that responds to the external (luminal) environment as well as internal stimuli by producing proteins that are secreted both apically and basolaterally. As exocrine cells that secrete proteins into the intestinal lumen, goblet cells play an important role in

Acknowledgements

The authors thank Elizabeth Brown for help in antibody production as well as Michelle Mikell and Alvan Chaney for germ-free and gnotobiotic mouse husbandry.

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^☆: Supported by National Institutes of Health grants DK47709 and AI39368 (to G.D.W.), DK49780 (to M.A.L.), AI37108 (to J.J.C.), and Center grant P30 DK-50306.

¹: W.H. and M.-L.W. contributed equally to this work.

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Basic-alimentary tractBacterial colonization leads to the colonic secretion of RELMβ/FIZZ2, a novel goblet cell-specific protein☆

Abstract

Background & Aims:

Methods:

Results:

Conclusions:

Section snippets

Animal studies

RELMβ is expressed in goblet cells

Discussion

Acknowledgements

Res Immunol

Semin Immunol

Surg Clin North Am

Gastroenterology

Gastroenterology

Gastroenterology

Am J Clin Nutr

J Biol Chem

J Biol Chem

Am J Clin Nutr

Gastroenterology

J Biol Chem

J Biol Chem

Trends Microbiol

J Biol Chem

Host-microbial symbiosis in the mammalian intestineexploring an internal ecosystem

Bioessays

A molecular sensor that allows a gut commensal to control its nutrient foundation in a competitive ecosystem

Proc Natl Acad Sci U S A

Molecular analysis of commensal host-microbial relationships in the intestine

Science

Prokaryotic regulation of epithelial responses by inhibition of IkappaB-alpha ubiquitination

Science

Functional biology of intestinal goblet cells

Am J Physiol

Intestinal mucins in colonization and host defense against pathogens

Am J Trop Med Hyg

Creating and maintaining the gastrointestinal ecosystemwhat we know and need to know from gnotobiology

Microbiol Mol Biol Rev

A family of tissue-specific resistin-like molecules

Proc Natl Acad Sci U S A

FIZZ1, a novel cysteine-rich secreted protein associated with pulmonary inflammation, defines a new gene family

EMBO J

The hormone resistin links obesity to diabetes

Nature

Cell type-specific expression and coregulation of murine resistin and resistin-like molecule-alpha in adipose tissue

Mol Endocrinol

Molecular analysis of the composition of the bifidobacterial and lactobacillus microflora of humans

Appl Environ Microbiol

Basic-alimentary tract
Bacterial colonization leads to the colonic secretion of RELMβ/FIZZ2, a novel goblet cell-specific protein☆