The insulin receptor gene and its expression in insulin-resistant mice

Defects of insulin receptor binding and tyrosine kinase activity have been described in genetically diabetic (db/db) and obese (ob/ob) mice. To determine if these changes were related to an abnormality in insulin receptor mRNA expression or structure of the receptor gene, we quantitated receptor mRNA from db/db and ob/ob homozygous, heterozygous (db/x, ob/x) and unaffected [db(x/x), ob(x/x)] mice and also analyzed restriction fragment length patterns of genomic DNA. Northern blot analysis of insulin receptor mRNA in livers from each of the genotypes revealed two major species of 7.5 and 9.5 kilobases. In contrast to known decreased receptor number in various tissues of ob/ob and db/db mice, quantitation of liver insulin receptor mRNA revealed that both homozygous affected strains had 2-fold or more increased levels of both major mRNA species compared to unaffected control groups. (P less than 0.05). Restriction fragment length analysis revealed no major insertion or deletion mutations in either the db/db or ob/ob insulin receptor gene. From the number and size of the fragments generated by this analysis, the minimal size of the mouse insulin receptor gene was calculated to be 97 kilobases, and the minimal number of exons was 16. These data indicate that the insulin receptor gene in ob/ob and db/db mice exhibits no major structural abnormality. Decreases in insulin receptor binding and/or kinase activity in affected mice appear to be due to a defect at the posttranscriptional level and occur despite increased levels of receptor mRNA.