Aim: The mechanisms underlying the age-related decrease in insulin-receptor (IR) binding in genetically obese Zucker rats are not well understood. For this reason, the present study analyzed the expression of IR mRNA and protein in selected tissues from 1- to 4-month-old obese (fa/fa) Zucker rats and lean (Fa/-) age-matched controls.
Methods: The following parameters were evaluated: (1) IR mRNA level, and proportion of isotypes A (exon 11-) and B (exon 11+) of IR mRNA in liver, brain and kidney; (2) level, molecular size and tyrosine phosphorylation of IR-beta subunit in liver subcellular fractions; and (3) stability of liver IR based on sensitivity in vivo of insulin-binding activity and IR-beta levels in response to tunicamycin, a glycosylation inhibitor.
Results: At one month, IR mRNA level was increased in liver and brain, but decreased in kidneys and, at four months, both mRNA level and isotype B proportion were decreased in liver. From age two months, the following changes in liver IR protein expression were observed: (1) decreased IR-beta level in whole homogenates, but increased IR-beta levels in endosomal fractions; (2) increased IR-beta tyrosine phosphorylation; and (3) at four months, increased levels of both intact IR-beta (95 kDa) and IR-beta fragments (72 and 52 kDa) in lysosomal fractions, along with decreased stability in vivo of the IR.
Conclusion: These data show that obese Zucker rats display age-related alterations of IR gene expression at both pre- and post-translational stages and, in particular, increased endocytosis and degradation of IR protein.
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