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1 Department of Health and Human Services, Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD, USA
* To whom correspondence should be addressed. E-mail: caiq{at}nhlbi.nih.gov.
In tests of osmotic tolerance of renal inner medullary cells in tissue culture, osmolality has usually been increased in a single step, whereas in vivo increase occurs gradually over hours. We previously found that more passage 2 mouse inner medullary epithelial (p2mIME) cells survive a linear increase of NaCl and urea from 640 to 1640 mosmol/kg over 20 hours (which is similar to the change that may occur in vivo) than they do a step increase. The present studies examine accompanying differences in gene expression. Among mRNAs of genes known to be protective, TonEBP and aldose reductase increase with linear, but decrease with step; BGT1 decreases with step, but not linear; HSP70.1 and HSP70.3 increase more with linear than step; OSP94 and HO-1 increase with linear, but decrease with step. mRNAs for known urea-responsive proteins, GADD153 and Egr-1 increase with both step and linear. Step increase of urea, alone, reduces mRNAs, similar to the combination of NaCl and urea, but step increase of NaCl, alone does not. HSP70 protein increases substantially with linear rise in osmolality, but does not change significantly with step rise. We speculate that poorer survival of p2mIME cells with step than with linear increase in NaCl and urea is accounted for, at least in part, by urea-induced suppression of protective genes, particularly HSP70.
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