Abstract
Inbred strains of mice carry either one of two alleles for the Ren-1 gene on chromosome 1; Ren-1c or Ren-1d. The Ren-1d allele is found in association with a tightly linked duplicated locus, Ren-2. The relative accumulations of renin transcripts derived from each locus were found to differ between mice with one or two renin loci in three extra renal tissues, the adrenal gland, testis, and sex accessory gland tissue. We find that the two Ren-1 alleles exhibit characteristic expression patterns in these tissues. Using genetic analyses and an allele-specific dideoxynucleotide primer extension assay we have further characterized the expression differences in these tissues. The results suggest that the allele-specific expression patterns observed for the Ren-1 genes are regulated by closely associated sequences in cis.
Highlights
Inbred strains of mice carry either one of two alleles for the Ren-1 gene on chromosome 1; Ren-1"or RenId
Ren-ldExpression in theAdrenal Gland and Testis--Results of a previous tissue survey byNorthern hybridizationanalysis indicated renin mRNA accumulation in theadult murine testis and adrenal gland in addition to the kidney and submandibular gland (SMG) of Ren-ld,Ren-2d mice (e.g. DBA/2) [3]
DBA/2hadfailed to detect renin mRNA in sexaccessory gland tissue [3], these results suggested another case of differential expression of the Ren-1 alleles
Summary
Nomenclature-The renin structural genes of the two-renin gene strains DBA/2 and SWR ardeesignated Ren-Zdand Ren-9. RNA from homogenized tissue fractions produced by either suspected that the situation in the adrenal gland and testis would be analogousto theSMG, where expression differences between strains with one versus two renin loci isthe result of elevated Ren-2d expression. The urea lysis buffer-PC1 or guanidine isothiocyanate methods was We used the locus-specificddNTP primer extension assay, purified by centrifugation over a 5.8 M CsCl pad a t 35,000 rpm for 18 to evaluate precisely whichrenin gene (Ren-ldor Ren-2d) was h in aBeckman SW-41 rotor. Expressed in the adrenal gland and testis of mice with two renin loci This assay is dependent upon the differential termination of reverse transcripts generated from an endlabeled oligonucleotideprimer that hybridizes well to. Samples were neutralized with 100 pl of 0.3 M sodium acetate, 0.1 M Tris, pH 5.0, and precipitated by the addition of 1ml of 95% ethanol, recovered, washed twice with 70%ethanol, and analyzed on 20% or 8%acrylamide, 7 M urea sequencing gels
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