Limits...
Deoxynivalenol affects the composition of the basement membrane proteins and influences en route the migration of CD16(+) cells into the intestinal epithelium.

Nossol C, Diesing AK, Kahlert S, Kersten S, Kluess J, Ponsuksili S, Hartig R, Wimmers K, Dänicke S, Rothkötter HJ - Mycotoxin Res (2013)

Bottom Line: The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells).Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells.Thus, DON affects the composition of the BM.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, Leipziger Strasse 44, 39120, Magdeburg, Germany, constanze.nossol@med.ovgu.de.

ABSTRACT
The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.

Show MeSH

Related in: MedlinePlus

Evaluation of the in vivo experiment. a Laminin staining of the BM (green). Continuous staining was not detected, and the unstained areas of the basement membrane represent the pores of the BM. The length of the unlabelled area was taken as the diameter of the pores, given that the pores are circular. The blue arrow shows how the length of the BM was measured. b CD16+ cells (red) in close contact to the BM (laminin-stained, green); a triple-stained picture, nuclei (DAPI, blue), b CD16+ cells (white arrow) and a CD16+ dendrite (yellow arrow) in the epithelium
© Copyright Policy - OpenAccess
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3824239&req=5

Fig1: Evaluation of the in vivo experiment. a Laminin staining of the BM (green). Continuous staining was not detected, and the unstained areas of the basement membrane represent the pores of the BM. The length of the unlabelled area was taken as the diameter of the pores, given that the pores are circular. The blue arrow shows how the length of the BM was measured. b CD16+ cells (red) in close contact to the BM (laminin-stained, green); a triple-stained picture, nuclei (DAPI, blue), b CD16+ cells (white arrow) and a CD16+ dendrite (yellow arrow) in the epithelium

Mentions: Epifluorescence microscopy was performed on a Zeiss Axioplan2, photographs were taken with an AxioCam HRc camera (Zeiss, Germany) and analysed with Axiovision software 4.8 (Zeiss). Six pictures were taken of each section. The pore size and the number of per 1,000 μm BM were determined based on the laminin staining (Fig. 1). At the same time intraepithelial CD16+ cells or dendrites/1,000 μm (CD16+ cells/dendrites in the lamina epithelialis mucosae) were counted in every section of the epithelium. Confocal microscopy was performed on a Leica SP2 confocal microscope (Leica, Heidelberg, Germany). Visualisation of three-dimensional structures was performed by rayetrace rendering of optical sections and rotation (VolumeJ plugin Fiji, NIH).Fig. 1


Deoxynivalenol affects the composition of the basement membrane proteins and influences en route the migration of CD16(+) cells into the intestinal epithelium.

Nossol C, Diesing AK, Kahlert S, Kersten S, Kluess J, Ponsuksili S, Hartig R, Wimmers K, Dänicke S, Rothkötter HJ - Mycotoxin Res (2013)

Evaluation of the in vivo experiment. a Laminin staining of the BM (green). Continuous staining was not detected, and the unstained areas of the basement membrane represent the pores of the BM. The length of the unlabelled area was taken as the diameter of the pores, given that the pores are circular. The blue arrow shows how the length of the BM was measured. b CD16+ cells (red) in close contact to the BM (laminin-stained, green); a triple-stained picture, nuclei (DAPI, blue), b CD16+ cells (white arrow) and a CD16+ dendrite (yellow arrow) in the epithelium
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824239&req=5

Fig1: Evaluation of the in vivo experiment. a Laminin staining of the BM (green). Continuous staining was not detected, and the unstained areas of the basement membrane represent the pores of the BM. The length of the unlabelled area was taken as the diameter of the pores, given that the pores are circular. The blue arrow shows how the length of the BM was measured. b CD16+ cells (red) in close contact to the BM (laminin-stained, green); a triple-stained picture, nuclei (DAPI, blue), b CD16+ cells (white arrow) and a CD16+ dendrite (yellow arrow) in the epithelium
Mentions: Epifluorescence microscopy was performed on a Zeiss Axioplan2, photographs were taken with an AxioCam HRc camera (Zeiss, Germany) and analysed with Axiovision software 4.8 (Zeiss). Six pictures were taken of each section. The pore size and the number of per 1,000 μm BM were determined based on the laminin staining (Fig. 1). At the same time intraepithelial CD16+ cells or dendrites/1,000 μm (CD16+ cells/dendrites in the lamina epithelialis mucosae) were counted in every section of the epithelium. Confocal microscopy was performed on a Leica SP2 confocal microscope (Leica, Heidelberg, Germany). Visualisation of three-dimensional structures was performed by rayetrace rendering of optical sections and rotation (VolumeJ plugin Fiji, NIH).Fig. 1

Bottom Line: The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells).Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells.Thus, DON affects the composition of the BM.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, Leipziger Strasse 44, 39120, Magdeburg, Germany, constanze.nossol@med.ovgu.de.

ABSTRACT
The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.

Show MeSH
Related in: MedlinePlus