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Received April 8, 2002
Accepted after revision May 10, 2002
1 Physiologisches Institut der Universität, 80336 München, Germany
2 Anatomische Anstalt der Universität, 80336 München, Germany
3 Physiologisches Institut der Universität, Pettenkoferstrasse 12, 80336 München, Germany
* To whom correspondence should be addressed. E-mail: FX.Beck{at}physiol.med.uni-muenchen.de.
Intracellular ionic strength may play an important role in regulating the expression of genes encoding osmolyte-accumulating molecules. To establish whether a strict relation exists between these variables, intracellular ionic strength (sum of Na+, Cl- and K+ concentrations) and the relative abundance of mRNA derived from various tonicity-sensitive genes was examined using electron microprobe analysis and Northern blots on primary cultures of rat papillary collecting duct (PCD) cells following acute or long-term alterations in medium tonicity. Hypertonic medium (450 mosmol kg-1) evoked an initial rise in intracellular ionic strength (269 ± 5 vs. 194 ± 7 mmol (kg wet weight (wt))-1 in isotonic controls; means ± S.E.M.), which subsequently declined gradually, and a significantly higher abundance of bgt1 (Na+- and Cl--dependent betaine transporter), smit (Na+/myo-inositol cotransporter), ar (aldose reductase) and osp94 (osmotic stress protein 94) mRNAs. Conversely, exposure to hypotonic medium (200 mosmol kg-1) for 12 h was associated with significantly reduced intracellular ionic strength (153 ± 4 mmol (kg wet wt)-1) and significantly reduced the abundance of smit and ar mRNAs. PCD cells preconditioned in hypotonic medium and re-exposed to isotonic medium showed significantly higher abundance of these mRNAs than isotonic controls, although the intracellular ionic strength did not differ. Two further tonicity-sensitive genes responded differently to medium tonicity: while the abundance of hsp70 (heat shock protein 70) mRNA increased significantly following both hypo- and hypertonic stress, inos (inducible nitric oxide synthase) mRNA abundance correlated inversely with medium tonicity. These findings support the view that the effect of intracellular ionic strength on the expression of bgt1, smit, ar and osp94 is modulated by additional factors such as cell volume, and that its effect on the pathways regulating hsp70 and inos is even more complex.
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