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Albuminuria is associated with too few glomeruli and too much testosterone.

Long DA, Kolatsi-Joannou M, Price KL, Dessapt-Baradez C, Huang JL, Papakrivopoulou E, Hubank M, Korstanje R, Gnudi L, Woolf AS - Kidney Int. (2013)

Bottom Line: Genetic variation and both pre- and postnatal environmental factors may affect albuminuria in humans.The transcripts encoded proteins involved in oxidation/reduction reactions, ion transport, and enzymes involved in detoxification.These proteins may represent novel biomarkers and even therapeutic targets for early kidney and cardiovascular disease.

View Article: PubMed Central - PubMed

Affiliation: Nephro-Urology Unit, UCL Institute of Child Health, London, UK. d.long@ucl.ac.uk

ABSTRACT
Normally, the glomerular filtration barrier almost completely excludes circulating albumin from entering the urine. Genetic variation and both pre- and postnatal environmental factors may affect albuminuria in humans. Here we determine whether glomerular gene expression in mouse strains with naturally occurring variations in albuminuria would allow identification of proteins deregulated in relatively 'leaky' glomeruli. Albuminuria increased in female B6 to male B6 to female FVB/N to male FVB/N mice, whereas the number of glomeruli/kidney was the exact opposite. Testosterone administration led to increased albuminuria in female B6 but not female FVB/N mice. A common set of 39 genes, many expressed in podocytes, were significantly differentially expressed in each of the four comparisons: male versus female B6 mice, male versus female FVB/N mice, male FVB/N versus male B6 mice, and female FVB/N versus female B6 mice. The transcripts encoded proteins involved in oxidation/reduction reactions, ion transport, and enzymes involved in detoxification. These proteins may represent novel biomarkers and even therapeutic targets for early kidney and cardiovascular disease.

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Expression of candidate genes in cultured podocytes. (a, b) RNA was collected from undifferentiated (Undiff) and differentiated (Diff) podocytes and PCR performed for all annotated genes found to be differentially expressed in the microarray analysis. Positive controls (+ve) of reactions consisted of mouse whole kidney RNA. Negative controls (−ve) consisted of reactions without cDNA template. Panel a comprises genes that were found to be expressed in cultured podocytes and panel b comprises transcripts that were not detected using this methodology. Representative pictures of phalloidin-stained undifferentiated (c) and differentiated (d) podocytes showing the extensive process formation characteristic of the in vivo podocyte phenotype. (c, d) Bar=50 μm.
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fig3: Expression of candidate genes in cultured podocytes. (a, b) RNA was collected from undifferentiated (Undiff) and differentiated (Diff) podocytes and PCR performed for all annotated genes found to be differentially expressed in the microarray analysis. Positive controls (+ve) of reactions consisted of mouse whole kidney RNA. Negative controls (−ve) consisted of reactions without cDNA template. Panel a comprises genes that were found to be expressed in cultured podocytes and panel b comprises transcripts that were not detected using this methodology. Representative pictures of phalloidin-stained undifferentiated (c) and differentiated (d) podocytes showing the extensive process formation characteristic of the in vivo podocyte phenotype. (c, d) Bar=50 μm.

Mentions: Assessed by reverse transcriptase–PCR (RT–PCR) (Figure 3a), over half of the set of transcripts identified by the array were expressed in a podocyte line,20 whereas others were not detected in the same cells (Figure 3b). Using protein lysates from isolated glomeruli, we undertook semiquantitative western blotting for CYP4A12A, a cytochrome P450, and aspartocyclase 3 (ACY3), respectively the proteins encoded by Cyp4a12a and Acy3. Reliable signals for HSD3B2 could not be obtained with available antibodies. Levels of CYP4A12A and ACY3 appeared markedly and reproducibly greatest in FVB/N males; lesser levels of these proteins were detected in B6 male glomeruli but they were barely detectable or undetectable in female glomeruli of either strain (Figure 4a). Immunohistochemistry of FVB/N male kidneys demonstrated CYP4A12A (Figure 4c–f) and HSD3B2 (Figure 4g–j) in the glomeruli; with some of the signal in podocytes as evidenced by double labeling with podoplanin. Furthermore, CYP4A12A was detected in western blots of both undifferentiated and differentiated cultured podocytes (Figure 4b). In tissue sections, ACY3 was predominantly immunolocalized in parietal glomerular epithelia, with a faint signal in glomerular tufts (Figure 4k–n); we did not detect ACY3 in western blots of cultured podocytes.


Albuminuria is associated with too few glomeruli and too much testosterone.

Long DA, Kolatsi-Joannou M, Price KL, Dessapt-Baradez C, Huang JL, Papakrivopoulou E, Hubank M, Korstanje R, Gnudi L, Woolf AS - Kidney Int. (2013)

Expression of candidate genes in cultured podocytes. (a, b) RNA was collected from undifferentiated (Undiff) and differentiated (Diff) podocytes and PCR performed for all annotated genes found to be differentially expressed in the microarray analysis. Positive controls (+ve) of reactions consisted of mouse whole kidney RNA. Negative controls (−ve) consisted of reactions without cDNA template. Panel a comprises genes that were found to be expressed in cultured podocytes and panel b comprises transcripts that were not detected using this methodology. Representative pictures of phalloidin-stained undifferentiated (c) and differentiated (d) podocytes showing the extensive process formation characteristic of the in vivo podocyte phenotype. (c, d) Bar=50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3674403&req=5

fig3: Expression of candidate genes in cultured podocytes. (a, b) RNA was collected from undifferentiated (Undiff) and differentiated (Diff) podocytes and PCR performed for all annotated genes found to be differentially expressed in the microarray analysis. Positive controls (+ve) of reactions consisted of mouse whole kidney RNA. Negative controls (−ve) consisted of reactions without cDNA template. Panel a comprises genes that were found to be expressed in cultured podocytes and panel b comprises transcripts that were not detected using this methodology. Representative pictures of phalloidin-stained undifferentiated (c) and differentiated (d) podocytes showing the extensive process formation characteristic of the in vivo podocyte phenotype. (c, d) Bar=50 μm.
Mentions: Assessed by reverse transcriptase–PCR (RT–PCR) (Figure 3a), over half of the set of transcripts identified by the array were expressed in a podocyte line,20 whereas others were not detected in the same cells (Figure 3b). Using protein lysates from isolated glomeruli, we undertook semiquantitative western blotting for CYP4A12A, a cytochrome P450, and aspartocyclase 3 (ACY3), respectively the proteins encoded by Cyp4a12a and Acy3. Reliable signals for HSD3B2 could not be obtained with available antibodies. Levels of CYP4A12A and ACY3 appeared markedly and reproducibly greatest in FVB/N males; lesser levels of these proteins were detected in B6 male glomeruli but they were barely detectable or undetectable in female glomeruli of either strain (Figure 4a). Immunohistochemistry of FVB/N male kidneys demonstrated CYP4A12A (Figure 4c–f) and HSD3B2 (Figure 4g–j) in the glomeruli; with some of the signal in podocytes as evidenced by double labeling with podoplanin. Furthermore, CYP4A12A was detected in western blots of both undifferentiated and differentiated cultured podocytes (Figure 4b). In tissue sections, ACY3 was predominantly immunolocalized in parietal glomerular epithelia, with a faint signal in glomerular tufts (Figure 4k–n); we did not detect ACY3 in western blots of cultured podocytes.

Bottom Line: Genetic variation and both pre- and postnatal environmental factors may affect albuminuria in humans.The transcripts encoded proteins involved in oxidation/reduction reactions, ion transport, and enzymes involved in detoxification.These proteins may represent novel biomarkers and even therapeutic targets for early kidney and cardiovascular disease.

View Article: PubMed Central - PubMed

Affiliation: Nephro-Urology Unit, UCL Institute of Child Health, London, UK. d.long@ucl.ac.uk

ABSTRACT
Normally, the glomerular filtration barrier almost completely excludes circulating albumin from entering the urine. Genetic variation and both pre- and postnatal environmental factors may affect albuminuria in humans. Here we determine whether glomerular gene expression in mouse strains with naturally occurring variations in albuminuria would allow identification of proteins deregulated in relatively 'leaky' glomeruli. Albuminuria increased in female B6 to male B6 to female FVB/N to male FVB/N mice, whereas the number of glomeruli/kidney was the exact opposite. Testosterone administration led to increased albuminuria in female B6 but not female FVB/N mice. A common set of 39 genes, many expressed in podocytes, were significantly differentially expressed in each of the four comparisons: male versus female B6 mice, male versus female FVB/N mice, male FVB/N versus male B6 mice, and female FVB/N versus female B6 mice. The transcripts encoded proteins involved in oxidation/reduction reactions, ion transport, and enzymes involved in detoxification. These proteins may represent novel biomarkers and even therapeutic targets for early kidney and cardiovascular disease.

Show MeSH
Related in: MedlinePlus