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This study aimed to investigate the tissue-specific role of the insulin-like growth factor 1 (IGF-I) on glucose homeostasis in the high-fatness selected Berlin Fat Mouse Inbred (BFMI) line. Therefore, the expression of different IGF-I transcripts and IGF-I protein, IGF-binding proteins, insulin as well as glucose tolerance was analyzed in BFMI in comparison with that in lean mice. In addition, dietary effects were investigated. The BFMI line showed normal blood glucose clearance on standard diet, but on high-fat diet the clearance was impaired, indicating the beginning of insulin resistance. Circulating IGF-I and insulin levels were elevated in BFMI than in lean mice on both diets along with a down-regulation of three IGF-I binding proteins in BFMI mice. Serum IGF-I levels corresponded with the expression pattern for both hepatic and one class II splice variants in reproductive adipose tissue, but not in muscle. High insulin and high IGF-I levels likely prevent BFMI mice from diabetes.