After three decades of physiological research, the precise nature of the genetic lesion in Zucker fatty (fa/fa) rats remains unknown. Several methods have been used to identify preobese rats to detect the earliest phenotypic effects of the fa mutation. Most of these methods have used phenotypic characteristics that are not reliable until the second week of life, when increased adiposity is already evident. We have used a restriction fragment length polymorphism (RFLP) for a human genomic DNA probe (VC85) that is tightly linked to the fa locus on rat chromosome 5 to genotype the F2 progeny of a Zucker (13M) x Brown Norway (BN) fa/+ F1 intercross. Sixty-four rats, comprising five litters, were killed at 5-6 wk of age. DNA was isolated either from tail at age 4-7 days (36 rats) or from organs at the time of death (28 rats). Adiposity was scored using inguinal fat pad weight as a percentage of body weight. RFLP analysis was > 99% accurate in identifying obese (fa/fa) rats. This molecular genetic method can be used to genotype fatty rats from an appropriate genetic cross at any age, even prenatally. Moreover, this method can distinguish heterozygous from homozygous littermates, enabling an analysis of gene dosage effects.
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