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Metalloprotease meprin beta in rat kidney: glomerular localization and differential expression in glomerulonephritis.

Oneda B, Lods N, Lottaz D, Becker-Pauly C, Stöcker W, Pippin J, Huguenin M, Ambort D, Marti HP, Sterchi EE - PLoS ONE (2008)

Bottom Line: In addition, the meprin beta staining pattern in the latter was reminiscent of immunostaining with the sheep anti-Fx1A antiserum, commonly used in PHN induction.Using Western blot and immunoprecipitation assays we demonstrated that meprin beta is recognized by Fx1A antiserum and may therefore represent an auto-antigen in PHN.This might point to an involvement of meprin beta in this form of glomerulonephritis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.

ABSTRACT
Meprin (EC 3.4.24.18) is an oligomeric metalloendopeptidase found in microvillar membranes of kidney proximal tubular epithelial cells. Here, we present the first report on the expression of meprin beta in rat glomerular epithelial cells and suggest a potential involvement in experimental glomerular disease. We detected meprin beta in glomeruli of immunostained rat kidney sections on the protein level and by quantitative RT-PCR of laser-capture microdissected glomeruli on the mRNA level. Using immuno-gold staining we identified the membrane of podocyte foot processes as the main site of meprin beta expression. The glomerular meprin beta expression pattern was altered in anti-Thy 1.1 and passive Heymann nephritis (PHN). In addition, the meprin beta staining pattern in the latter was reminiscent of immunostaining with the sheep anti-Fx1A antiserum, commonly used in PHN induction. Using Western blot and immunoprecipitation assays we demonstrated that meprin beta is recognized by Fx1A antiserum and may therefore represent an auto-antigen in PHN. In anti-Thy 1.1 glomerulonephritis we observed a striking redistribution of meprin beta in tubular epithelial cells from the apical to the basolateral side and the cytosol. This might point to an involvement of meprin beta in this form of glomerulonephritis.

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Double immunofluorescence staining of kidney sections from Wistar rats.Immunofluorescence staining of glomeruli: (A) anti-meprinβ antibody, (B) anti-synaptopodin antibody, (C) merged image meprin/synaptopodin, (D) anti-meprinβ antibody, (E) anti-Thy 1.1 antibody, (F) merged image meprin/Thy 1.1. Nuclei are marked by staining with 4,6-diamidino-2-phenylindole (DAPI).
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pone-0002278-g002: Double immunofluorescence staining of kidney sections from Wistar rats.Immunofluorescence staining of glomeruli: (A) anti-meprinβ antibody, (B) anti-synaptopodin antibody, (C) merged image meprin/synaptopodin, (D) anti-meprinβ antibody, (E) anti-Thy 1.1 antibody, (F) merged image meprin/Thy 1.1. Nuclei are marked by staining with 4,6-diamidino-2-phenylindole (DAPI).

Mentions: To further investigate the localization of meprinβ in glomeruli, we double-stained the Wistar kidney sections with anti-meprinβ antibodies and anti-synaptopodin or anti-Thy 1.1 antibodies (Fig. 2). Synaptopodin is abundantly expressed in the foot processes of podocytes whereas Thy 1.1 is localized on the surface of glomerular mesangial cells. The staining pattern of meprinβ revealed co-localization with synaptopodin (Fig. 2A–C), but not with Thy 1.1 (Fig. 2D–F). Both meprinβ and synaptopodin displayed a linear staining pattern drawing the contour of the intra-glomerular capillaries. These findings clearly demonstrate the expression of meprinβ protein in podocytes of rat glomeruli.


Metalloprotease meprin beta in rat kidney: glomerular localization and differential expression in glomerulonephritis.

Oneda B, Lods N, Lottaz D, Becker-Pauly C, Stöcker W, Pippin J, Huguenin M, Ambort D, Marti HP, Sterchi EE - PLoS ONE (2008)

Double immunofluorescence staining of kidney sections from Wistar rats.Immunofluorescence staining of glomeruli: (A) anti-meprinβ antibody, (B) anti-synaptopodin antibody, (C) merged image meprin/synaptopodin, (D) anti-meprinβ antibody, (E) anti-Thy 1.1 antibody, (F) merged image meprin/Thy 1.1. Nuclei are marked by staining with 4,6-diamidino-2-phenylindole (DAPI).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2386549&req=5

pone-0002278-g002: Double immunofluorescence staining of kidney sections from Wistar rats.Immunofluorescence staining of glomeruli: (A) anti-meprinβ antibody, (B) anti-synaptopodin antibody, (C) merged image meprin/synaptopodin, (D) anti-meprinβ antibody, (E) anti-Thy 1.1 antibody, (F) merged image meprin/Thy 1.1. Nuclei are marked by staining with 4,6-diamidino-2-phenylindole (DAPI).
Mentions: To further investigate the localization of meprinβ in glomeruli, we double-stained the Wistar kidney sections with anti-meprinβ antibodies and anti-synaptopodin or anti-Thy 1.1 antibodies (Fig. 2). Synaptopodin is abundantly expressed in the foot processes of podocytes whereas Thy 1.1 is localized on the surface of glomerular mesangial cells. The staining pattern of meprinβ revealed co-localization with synaptopodin (Fig. 2A–C), but not with Thy 1.1 (Fig. 2D–F). Both meprinβ and synaptopodin displayed a linear staining pattern drawing the contour of the intra-glomerular capillaries. These findings clearly demonstrate the expression of meprinβ protein in podocytes of rat glomeruli.

Bottom Line: In addition, the meprin beta staining pattern in the latter was reminiscent of immunostaining with the sheep anti-Fx1A antiserum, commonly used in PHN induction.Using Western blot and immunoprecipitation assays we demonstrated that meprin beta is recognized by Fx1A antiserum and may therefore represent an auto-antigen in PHN.This might point to an involvement of meprin beta in this form of glomerulonephritis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.

ABSTRACT
Meprin (EC 3.4.24.18) is an oligomeric metalloendopeptidase found in microvillar membranes of kidney proximal tubular epithelial cells. Here, we present the first report on the expression of meprin beta in rat glomerular epithelial cells and suggest a potential involvement in experimental glomerular disease. We detected meprin beta in glomeruli of immunostained rat kidney sections on the protein level and by quantitative RT-PCR of laser-capture microdissected glomeruli on the mRNA level. Using immuno-gold staining we identified the membrane of podocyte foot processes as the main site of meprin beta expression. The glomerular meprin beta expression pattern was altered in anti-Thy 1.1 and passive Heymann nephritis (PHN). In addition, the meprin beta staining pattern in the latter was reminiscent of immunostaining with the sheep anti-Fx1A antiserum, commonly used in PHN induction. Using Western blot and immunoprecipitation assays we demonstrated that meprin beta is recognized by Fx1A antiserum and may therefore represent an auto-antigen in PHN. In anti-Thy 1.1 glomerulonephritis we observed a striking redistribution of meprin beta in tubular epithelial cells from the apical to the basolateral side and the cytosol. This might point to an involvement of meprin beta in this form of glomerulonephritis.

Show MeSH
Related in: MedlinePlus