International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


M Inoue
RIKEN Genomic Sciences Center (GSC)

1) Motegi H, 2) Kubota N, 1) Matsui J, 1) Toki H, 1) Adachi T, 1) Kagami T, 1) Inoue A, 1) Takahashi Y, 3) Shigeyama Y, 1) Kobayashi K, 1) Wakana S, 1) Gondo Y, 1) Minowa O, 1) Shiroishi T, 1) Noda T.
1) RIKEN GSC, 2) University of Tokyo, 3) Kobe University

To generate mouse models of human diabetes, we utilized clinical biochemistry test in the course of ENU-mutagenesis procedure as the first step of mutant screening. At 11 weeks of age 200 ul of blood was collected from G1 animals (offspring of ENU-mutagenized male mice) and 75 ul of its serum was analyzed by automatic analyser JCA-BM2250 (JEOL). About 8,500 G1 animals were screened, where 22 animals showed high blood glucose, and 18 were entered into inheritance testing. Of these 18, 7 lines were confirmed as inherited, 6 were found not to be inherited and 5 are still being tested. To detect more detailed phenotypes of each mutant line, oral glucose tolerance tests (OGTT) and insulin tolerance tests (ITT) were performed for the mutant founders offspring. Before the OGTT, mice were fasted for 16-18 hrs and then given 1.5 mg of glucose/g of body weight. Blood glucose and/or plasma insulin levels were measured at 0, 30, 60 and 120 min after glucose administration. For the ITT, freely fed mice were given 0.75 mU of human insulin/g of body weight. Blood glucose levels were measured at 0, 20, 40, 60, 80, 100 and 120 min after insulin administration. With these tests we started to clarify differences in insulin sensitivity and/or secretion among these mutants. Our results have suggested that the biochemical screen in ENU mutagenesis is an efficient tool to establish novel and various diabetes models. Newly isolated mutants currently under characterization will be presented and their features will be discussed.

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