Use of gene chip technology for the characterisation of the regulation of renal transport processes and of nephrotoxicity in rats.
Gene expression profiling using microarrays (rat-specific array RG-U34A, Affymetrix, U.S.A.) was employed for the investigation of: (1) hormonal regulation of renal function and (2) nephrotoxicity. For this purpose about 8,800 genes were analysed in kidney and, additionally, in liver tissue. Ad 1.) Kidney functions develop during postnatal life. Thus, in vivo transport and accumulation of p-aminohippurate (PAH) was investigated on renal cortical slices (RCS) from 10- and 55-day-old rats. The animals were treated with dexamethasone (DEXA; 60 microg/100 g b.wt./day) for 3 days, which caused a significant reduction in the accumulation of PAH in 10-day-old rats (42 +/- 5% whereas it was only slightly reduced in 55-day-old rats (70 +/- 8%). To further clarify the regulation of renal function by DEXA, results were compared with those obtained previously after in vitro stimulation with DEXA. RCS were incubated for 24 hours in DEXA-containing medium (10(-9) M). Under these conditions DEXA significantly increased the PAH uptake capacity in RCS obtained from 10- and 55-day-old rats up to 126 and 136%, respectively. Thus a stimulation of tubular transport capacity is possible in vitro. The effect of DEXA treatment on the gene expression of the kidney (in vivo) was moderate. Focussing especially on transporters, ion channels, ATPases, glucuronyltransferases, glutathione-S-transferase and cytochrome P450, the expression of only few genes were significantly changed (3 to 50-fold up- or down-regulation). Moreover, distinct age differences were found after in vivo administration of DEXA. The investigation of in vitro effects of DEXA is currently been performed. Ad 2.) The kidney is threatened by nephrotoxins because of its ability to accumulate them. We used a single administration of uranyl nitrate (UN; 0.5 mg/100 g b.wt.) as a model for chronic renal failure (CRF). Clearance experiments were performed 10 weeks after UN administration (maximal symptoms of CRF) in adult female rats. As expected, UN induced interstitial cicatrices with reduced GFR and diminished PAH transport capacity. Despite the impressive morphological and functional changes in the kidney after exposure to UN, the gene expression profiles in the kidneys were only minimally affected: we found significantly changed expression levels for only 20 genes (5 genes were up-regulated [e.g. transgelin], 15 down-regulated [among these the Na-K-Cl-symporter, insulin-like growth factor, kallikrein, and ornithine decarboxylase). The lack of agreement between gene expression data and the nephrotoxic effects of UN can probably be explained by the long time interval between dosing and the assessment of the effect. The results confirm that primary genomic responses are likely to be strongest transiently after exposure and then decrease in intensity.
Fleck
Christian
C
Institute of Pharmacology and Toxicology, Friedrich Schiller University Jena, Germany. Christian.Fleck@mti.uni-jena.de
Sutter
Laura
L
Appenroth
Dorothea
D
Koch
Berit
B
Meinhold
Tobias
T
Pitack
Melanie
M
Gasser
Rodolfo
R
D000375:Aging; D000818:Animals; D001692:Biological Transport; D003907:Dexamethasone / Q000494:pharmacology; D004195:Disease Models, Animal; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007668:Kidney / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D007672:Kidney Cortex / Q000187:drug effects / Q000378:metabolism; D007676:Kidney Failure, Chronic / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D014502:Uranyl Nitrate / Q000633:toxicity; D010130:p-Aminohippuric Acid / Q000378:metabolism
D014502:Uranyl Nitrate; D003907:Dexamethasone; D010130:p-Aminohippuric Acid