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Systematic analysis of a novel human renal glomerulus-enriched gene expression dataset.

Lindenmeyer MT, Eichinger F, Sen K, Anders HJ, Edenhofer I, Mattinzoli D, Kretzler M, Rastaldi MP, Cohen CD - PLoS ONE (2010)

Bottom Line: This analysis resulted in 677 genes with prominent overrepresentation in the glomerulus.Gene ontology and pathway analysis identified biological processes previously not reported to be of relevance in glomeruli of healthy human adult kidneys including among others axon guidance.In diabetic nephropathy, a prevalent glomerulopathy, differential regulation of glomerular ROBO2 mRNA was found.In summary, novel transcripts with predominant expression in the human glomerulus could be identified using a comparative strategy on microdissected nephrons.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, University Hospital Zurich, Zurich, Switzerland.

ABSTRACT
Glomerular diseases account for the majority of cases with chronic renal failure. Several genes have been identified with key relevance for glomerular function. Quite a few of these genes show a specific or preferential mRNA expression in the renal glomerulus. To identify additional candidate genes involved in glomerular function in humans we generated a human renal glomerulus-enriched gene expression dataset (REGGED) by comparing gene expression profiles from human glomeruli and tubulointerstitium obtained from six transplant living donors using Affymetrix HG-U133A arrays. This analysis resulted in 677 genes with prominent overrepresentation in the glomerulus. Genes with 'a priori' known prominent glomerular expression served for validation and were all found in the novel dataset (e.g. CDKN1, DAG1, DDN, EHD3, MYH9, NES, NPHS1, NPHS2, PDPN, PLA2R1, PLCE1, PODXL, PTPRO, SYNPO, TCF21, TJP1, WT1). The mRNA expression of several novel glomerulus-enriched genes in REGGED was validated by qRT-PCR. Gene ontology and pathway analysis identified biological processes previously not reported to be of relevance in glomeruli of healthy human adult kidneys including among others axon guidance. This finding was further validated by assessing the expression of the axon guidance molecules neuritin (NRN1) and roundabout receptor ROBO1 and -2. In diabetic nephropathy, a prevalent glomerulopathy, differential regulation of glomerular ROBO2 mRNA was found.In summary, novel transcripts with predominant expression in the human glomerulus could be identified using a comparative strategy on microdissected nephrons. A systematic analysis of this glomerulus-specific gene expression dataset allows the detection of target molecules and biological processes involved in glomerular biology and renal disease.

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Immunofluorescence of robo1 (A) and neuritin (B) in human control kidneys.Immunofluorescence for robo1 (A) shows a constitutive expression in glomeruli, while there is no expression in the tubulointerstitium. Immunostaining for neuritin (B) shows a clear glomerular expression associated to some positivity in the tubulointerstitium. The hatched line displays the glomerular contour (indirect immunofluorescence, DAPI nuclear counterstain, 200X).
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pone-0011545-g006: Immunofluorescence of robo1 (A) and neuritin (B) in human control kidneys.Immunofluorescence for robo1 (A) shows a constitutive expression in glomeruli, while there is no expression in the tubulointerstitium. Immunostaining for neuritin (B) shows a clear glomerular expression associated to some positivity in the tubulointerstitium. The hatched line displays the glomerular contour (indirect immunofluorescence, DAPI nuclear counterstain, 200X).

Mentions: To evaluate the protein expression of both axon guidance molecules in vivo, immunofluorescence staining for robo1 and neuritin was performed on an independent set of healthy control kidneys. For robo1 expression in the glomerulus could be detected, while the staining was either completely absent or showed a minimal scattered positivity in the tubulointerstitial compartment. Neuritin showed a clear staining of the glomerulus, but also some positive signal in the tubulointerstitium (Figure 6A and B). This is consistent with REGGED and the qRT-PCR experiments indicating an enhanced but not specific glomerular expression of neuritin in the human adult healthy kidney.


Systematic analysis of a novel human renal glomerulus-enriched gene expression dataset.

Lindenmeyer MT, Eichinger F, Sen K, Anders HJ, Edenhofer I, Mattinzoli D, Kretzler M, Rastaldi MP, Cohen CD - PLoS ONE (2010)

Immunofluorescence of robo1 (A) and neuritin (B) in human control kidneys.Immunofluorescence for robo1 (A) shows a constitutive expression in glomeruli, while there is no expression in the tubulointerstitium. Immunostaining for neuritin (B) shows a clear glomerular expression associated to some positivity in the tubulointerstitium. The hatched line displays the glomerular contour (indirect immunofluorescence, DAPI nuclear counterstain, 200X).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2902524&req=5

pone-0011545-g006: Immunofluorescence of robo1 (A) and neuritin (B) in human control kidneys.Immunofluorescence for robo1 (A) shows a constitutive expression in glomeruli, while there is no expression in the tubulointerstitium. Immunostaining for neuritin (B) shows a clear glomerular expression associated to some positivity in the tubulointerstitium. The hatched line displays the glomerular contour (indirect immunofluorescence, DAPI nuclear counterstain, 200X).
Mentions: To evaluate the protein expression of both axon guidance molecules in vivo, immunofluorescence staining for robo1 and neuritin was performed on an independent set of healthy control kidneys. For robo1 expression in the glomerulus could be detected, while the staining was either completely absent or showed a minimal scattered positivity in the tubulointerstitial compartment. Neuritin showed a clear staining of the glomerulus, but also some positive signal in the tubulointerstitium (Figure 6A and B). This is consistent with REGGED and the qRT-PCR experiments indicating an enhanced but not specific glomerular expression of neuritin in the human adult healthy kidney.

Bottom Line: This analysis resulted in 677 genes with prominent overrepresentation in the glomerulus.Gene ontology and pathway analysis identified biological processes previously not reported to be of relevance in glomeruli of healthy human adult kidneys including among others axon guidance.In diabetic nephropathy, a prevalent glomerulopathy, differential regulation of glomerular ROBO2 mRNA was found.In summary, novel transcripts with predominant expression in the human glomerulus could be identified using a comparative strategy on microdissected nephrons.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, University Hospital Zurich, Zurich, Switzerland.

ABSTRACT
Glomerular diseases account for the majority of cases with chronic renal failure. Several genes have been identified with key relevance for glomerular function. Quite a few of these genes show a specific or preferential mRNA expression in the renal glomerulus. To identify additional candidate genes involved in glomerular function in humans we generated a human renal glomerulus-enriched gene expression dataset (REGGED) by comparing gene expression profiles from human glomeruli and tubulointerstitium obtained from six transplant living donors using Affymetrix HG-U133A arrays. This analysis resulted in 677 genes with prominent overrepresentation in the glomerulus. Genes with 'a priori' known prominent glomerular expression served for validation and were all found in the novel dataset (e.g. CDKN1, DAG1, DDN, EHD3, MYH9, NES, NPHS1, NPHS2, PDPN, PLA2R1, PLCE1, PODXL, PTPRO, SYNPO, TCF21, TJP1, WT1). The mRNA expression of several novel glomerulus-enriched genes in REGGED was validated by qRT-PCR. Gene ontology and pathway analysis identified biological processes previously not reported to be of relevance in glomeruli of healthy human adult kidneys including among others axon guidance. This finding was further validated by assessing the expression of the axon guidance molecules neuritin (NRN1) and roundabout receptor ROBO1 and -2. In diabetic nephropathy, a prevalent glomerulopathy, differential regulation of glomerular ROBO2 mRNA was found.In summary, novel transcripts with predominant expression in the human glomerulus could be identified using a comparative strategy on microdissected nephrons. A systematic analysis of this glomerulus-specific gene expression dataset allows the detection of target molecules and biological processes involved in glomerular biology and renal disease.

Show MeSH
Related in: MedlinePlus