Impaired beta-adrenergic signaling pathway in white adipocytes of suckling fa/fa Zucker rats: a defect in receptor coupling.

Abstract

In fa/fa Zucker rats, leptin receptor deficiency is responsible for both a deficit of energy expenditure and hyperphagia which lead to massive obesity and insulin resistance in adulthood. This obesity is also characterised by alterations of the beta-adrenergic signaling pathway. To determine whether alterations in beta-adrenergic pathway could occur at the onset of obesity when fa/fa rats are not yet hyperinsulinemic. Fourteen-day-old suckling fa/fa and Fa/fa littermates (from heterozygous lean (Fa/fa) female and homozygous obese (fa/fa) male mating). Membranes were prepared from isolated adipocytes after collagenase treatment of inguinal adipose tissue. The response of adenylyl-cyclase activity to stimulation by isoprenaline, GTPgamma-S or forskolin was studied. Bmax and Kd of (beta1+beta2) and of beta3 adrenoceptors were measured using 3H-CGP saturation binding experiments. mRNA concentration of beta1- and beta3-AR was determined by semi-quantitative RT-PCR. G(s)alpha protein was quantified by Western blotting and Gi protein by ADP-ribosylation. Despite an almost normal body weight, inguinal fat pad weight was increased two-fold by the expression of fa mutation. This increase was entirely accounted for by fat cell hypertrophy (x2.5 in volume). In fa/fa compared to Fa/fa pups, response of adenylyl cyclase to isoprenaline was decreased two-fold but responses to GTPgammaS or forskolin were unchanged. Density of (beta1+beta2) and beta3-AR was not affected by the fa/fa genotype, as well as G(s)alpha and Gi concentration. Response of inguinal fat cells to catecholamines was decreased without any quantitative modifications of the different elements of the adenylyl cyclase cascade. This suggests an alteration in the coupling between beta-AR and G proteins. Due to the important increase in fat cell volume we hypothesize that changes in the physical properties of plasma membranes and/or changes in cytoskeleton-extracellular-matrix interactions could disturb the beta-adrenergic pathway responsiveness. In addition to the excess of lipid storage, which occurs very early at the onset of obesity, the impairment of the responsiveness to catecholamines reported in this study might worsen the obesity syndrome.