Genetic control and genotyping of Zucker (fa/+) rats for the maintenance of the lineage of animal models for the study of human Diabetes. Amanda B. Pustiglione, Clélia R. A. Bertoncini, Daniel S. Vilas Boas

Abstract

Diabetes mellitus is a chronic disease in which many of its complications make the individual unable to perform daily activities. It is generally classified into two types: Type 1, which is characterized by absolute insulin deficiency due to destruction of pancreatic β cells, and Type 2, which is caused by a combination of peripheral insulin resistance and an inadequate secretory response of β cells. It is known that no animal model of diabetes presents 100% identity to the human syndrome, but Zucker rats have shown satisfactory results. The present study aimed to create and maintain a line of animal models for the study of human diabetes through molecular techniques of genetic control and genotyping of Zucker rats (fa/+). The genotyping protocol was adapted from previously established methodologies and performed in a PCR thermocycler. After the reactions, the PCR products were subjected to agarose gel electrophoresis in TBE buffer containing ethidium bromide, using a 100bp DNA standard. To confirm the identity, the PCR amplified products were subjected to digestion by the restriction enzyme Msp I. Through this study we concluded that of the 70 genotyped neonates, 20% were born normal, and 77.1% were heterozygous (they have the mutation). And that these 77.1% should be used for the maintenance of the lineage, separating by sex, for mating.