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Evidence for a differential expression of fibronectin splice forms ED-A and ED-B in Crohn’s disease (CD) mucosa

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International Journal of Colorectal Disease Aims and scope Submit manuscript

Abstract

Background and aims

Fibronectin (FN) is an essential factor for the induction of migration of primary colonic lamina propria fibroblasts (CLPF). The FN isoform ED-A is an important inducer of migration. Recently, we have shown that CLPF isolated from inflamed Crohn’s disease (CD) mucosa migrated significantly less than control CLPF. We, therefore, investigated changes in FN or integrin expression that could be relevant for CLPF migration.

Materials and methods

mRNA of control-CLPF and CLPF isolated from fibrotic mucosa of CD patients was subtractively hybridized. Expression of FN, ED-A, and ED-B in frozen sections from intestinal mucosa was determined by immunohistochemistry. The mRNA expression of the FN isoforms in control, CD, and fibrosis biopsies was quantified by real-time polymerase chain reaction (PCR). Integrin α5β1 protein and mRNA expression was analyzed by fluorescence activated cell sorting (FACS) and PCR, respectively.

Results

Subtractive hybridization indicated differential regulation of FN isoform expression in CD. The immunohistochemical analysis of FN protein revealed a reduction of FN isoforms in inflamed CD mucosa compared to control mucosa. In CD fistulae, the ED-A and ED-B isoforms were virtually absent. In fibrotic mucosa, both proteins were increased. Real-time PCR showed a decrease of FN and ED-A expression during mucosal inflammation in CD in contrast to UC and a significant increase of FN and isoforms in CD fibrosis. No difference was found for protein and mRNA of integrin α5β1 in control, CD, and fibrosis CLPF by FACS and PCR.

Conclusion

Downregulated expression of migration-inducing FN-isoforms in contrast to unchanged FN receptor expression may contribute to the observed alterations of CD CLPF migration.

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Abbreviations

FN:

Fibronectin

IBD:

Inflammatory bowel disease

CLPF:

Colonic lamina propria fibroblasts

CD:

Crohn’s disease

ED-A:

Extra domain A

ED-B:

Extra domain B

IIICS:

Type III connecting segment

EDTA:

Ethylendiaminetetraacetic acid

PBS:

Phosphate buffered saline

DMEM:

Dulbecco’s Modified Eagle’s medium

FCS:

Fetal calf serum

SDS:

Sodium dodecyl sulfate

SSC:

Sodium chloride–sodium citrate buffer

GAPDH:

Glycerinaldehyde-3-phosphate dehydrogenase

VEGF:

Vascular endothelial growth factor

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Authors and Affiliations

  1. Department of Internal Medicine I, University of Regensburg, 93042, Regensburg, Germany

    Julia Brenmoehl, Markus Lang, Martin Hausmann, Sandra N. Leeb, Werner Falk, Jürgen Schölmerich, Michael Göke & Gerhard Rogler

  2. Department of Internal Medicine, Malteser Hospital, Bonn-Hardtberg, Germany

    Michael Göke

Authors
  1. Julia Brenmoehl
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  2. Markus Lang
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  3. Martin Hausmann
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  4. Sandra N. Leeb
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  6. Jürgen Schölmerich
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  7. Michael Göke
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  8. Gerhard Rogler
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Corresponding author

Correspondence to Gerhard Rogler.

Additional information

Supported by the BMBF “Kompetenznetzwerk-Chronisch entzündliche Darmerkrankungen” Parts of this manuscript were presented at DDW 2004 and DDW 2005.

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Brenmoehl, J., Lang, M., Hausmann, M. et al. Evidence for a differential expression of fibronectin splice forms ED-A and ED-B in Crohn’s disease (CD) mucosa. Int J Colorectal Dis 22, 611–623 (2007). https://doi.org/10.1007/s00384-006-0188-4

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