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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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Western Blot analysis of robo1 (A) and neuritin (B) in human podocytes.For robo1 (A) and neuritin (nrn1) (B), a band of the expected size was found in a human podocyte cell line; human brain lysate served as a positive control. Beta-actin was used as an internal loading control.
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pone-0011545-g004: Western Blot analysis of robo1 (A) and neuritin (B) in human podocytes.For robo1 (A) and neuritin (nrn1) (B), a band of the expected size was found in a human podocyte cell line; human brain lysate served as a positive control. Beta-actin was used as an internal loading control.

Mentions: The protein expression of neuritin and roundabout receptor 1, both known to be involved in axon guidance and neurite outgrowth, was assessed by Western blot in podocytes. Both proteins, robo1 and neuritin, were found to be expressed on protein level in a human podocyte cell line; human brain lysate served as a positive control (Figure 4A and B).


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)

Western Blot analysis of robo1 (A) and neuritin (B) in human podocytes.For robo1 (A) and neuritin (nrn1) (B), a band of the expected size was found in a human podocyte cell line; human brain lysate served as a positive control. Beta-actin was used as an internal loading control.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0011545-g004: Western Blot analysis of robo1 (A) and neuritin (B) in human podocytes.For robo1 (A) and neuritin (nrn1) (B), a band of the expected size was found in a human podocyte cell line; human brain lysate served as a positive control. Beta-actin was used as an internal loading control.
Mentions: The protein expression of neuritin and roundabout receptor 1, both known to be involved in axon guidance and neurite outgrowth, was assessed by Western blot in podocytes. Both proteins, robo1 and neuritin, were found to be expressed on protein level in a human podocyte cell line; human brain lysate served as a positive control (Figure 4A and B).

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