The basic helix-loop-helix (bHLH) class of proteins are of major importance in controlling tissue-specific gene expression. The actions of the bHLH proteins are inhibited by a related class of proteins, inhibitors of differentiation (Id). We have studied the expression of one of these latter proteins, Id-1, in the normal and post-ischemic regenerating rat kidney by immunocytochemistry, Western blot and RNase protection assay (RPA) and correlated Id-1 regulation to the expression of vimentin and proliferating cellular nuclear antigen (PCNA). In the normal kidney strong immunostaining for Id-1 was found in the distal nephron, especially in the distal convoluted tubule in the cortex. In particular, the perinuclear region was intensely stained in the cells of the distal tubule. mRNA for Id-1I was detectable by RPA on total RNA extracted from the renal cortex of sham-operated animals. The Id-1 monomer was detected on Western blots of normal animals. Vimentin was expressed in the mesangial cells of the glomeruli and in cells in the interstitium while tubule cells were negative. The labeling intensity for PCNA was low in all cellular compartments in the normal kidney. In the regenerating kidneys at various time intervals, the expression of Id-1-like immunoreactivity was widespread in the regenerating dedifferentiated tubule cells while by the end of the study period, more highly differentiated tubule cells appeared to lose their staining. On Western blots the Id-1 monomer was undetectable and instead strong staining was seen in the high molecular range. Id-1 mRNA levels in the regenerating kidneys did not differ significantly when compared to sham. PCNA labeling was intense in the regenerating kidneys at all time periods studied, indicating the intense proliferative activity in the regenerating kidneys. Vimentin expression in the renal tubule cells was increased from day 3 and onward. The data are consistent with a hypothesis in which Id-1 regulates differentiation of renal tubule cells in the post-ischemic regenerating rat kidney.
Animals Blotting, Western