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      Expression of Id-1 mRNA and Protein in the Post-Ischemic Regenerating Rat Kidney

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          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-1 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.

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            The daughterless gene product in Drosophila is a nuclear protein that is broadly expressed throughout the organism during development.

            The daughterless (da) gene in Drosophila functions in the regulation of at least three significant developmental pathways: sex determination, neurogenesis and oogenesis. As a member of the helix-loop-helix (HLH) family of DNA binding proteins, the da gene product appears to act as a transcription factor. Based on the genetic and molecular characterization of da, it has been proposed that the da protein (Da) functions as a generic member of this family, serving throughout development as a necessary binding partner for an assortment of other HLH proteins. As a result of temporally and/or spatially restricted expression, these binding partners would provide some regulatory specificity to the functional transcription complex. In order to participate in this way in the regulation of multiple genes, Da must be expressed in numerous times and places during development. Using anti-Da antibodies, we validate two predictions of this scenario of Da function: (1) Da protein is not only nuclear localized, but also associated with chromosomes in vivo; and (2) Da protein is widely distributed, both spatially and temporally, throughout development. With regard to the essential role of maternal da+ in progeny sex determination, little, if any, Da protein is synthesized in the maternal germline. This suggests that the female-specific germline function of da+ is provided to the zygote as maternally synthesized RNA that becomes translated early in embryogenesis.
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              Structural organisation and chromosomal mapping of the human Id-3 gene.

              The helix-loop-helix (HLH) family of transcription factors plays a central role in the regulation of cell growth, differentiation and tumourigenesis. Members of the Id (inhibitor of DNA binding) class of these nuclear proteins are able to heterodimerise with and thereby antagonise the functions of other transcription factors of this family. We report here on the genomic organisation of the human Id3 (HLH 1R21/heir1) gene. Comparison with the two other mammalian Id genes, Id1 and Id2, reveals a highly conserved protein coding gene organisation consistent with evolution from a common, ancestral Id-like gene. In addition, by using a yeast artificial chromosome (YAC) clone of Id3, we have fine-scale mapped the gene to chromosome band 1p36.1 by fluorescence in situ hybridisation (FISH) and, using the same FISH technique, we have detected heterogeneity in tumour-associated 1p36 chromosome translocations.

                Author and article information

                Nephron Exp Nephrol
                Cardiorenal Medicine
                S. Karger AG
                June 1998
                22 May 1998
                : 6
                : 3
                : 253-264
                a Research Center for Endocrinology and Metabolism, University of Göteborg, and b Department of Internal Medicine, Division of Nephrology, and c Department of Clinical Chemistry, Sahlgrenska Hospital, Göteborg, Sweden
                20530 Exp Nephrol 1998;6:253–264
                © 1998 S. Karger AG, Basel

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                Page count
                Figures: 4, Tables: 1, References: 46, Pages: 12
                Self URI (application/pdf): https://www.karger.com/Article/Pdf/20530
                Original Paper

                Cardiovascular Medicine, Nephrology

                Regeneration, Kidney, bHLH, Id-1


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