Characterization of nuclear corticosteroid receptors in rat adipocytes. Regional variations and modulatory effects of hormones.
The corticosteroid receptor was investigated in isolated rat adipocytes with a new technique which characterizes the corticosteroid receptors that can be activated and tightly bound to the nucleus. The binding reaction with [3H]triamcinolone was performed with intact isolated adipocytes and the radioactivity associated with nucleus was subsequently determined after cell lysis. Scatchard analysis revealed a homogeneous class of nuclear corticosteroid receptors in rat epididymal adipocytes with an apparent Kd of 4.93 +/- 1.5 nM and a Bmax of 21.8 +/- 6.6 fmol/10(6) cells corresponding to about 13,000 receptors per nucleus. The corticosteroid binding exhibited regional variations in isolated adipocytes. The highest receptor number was found in epididymal adipocytes (Bmax 25.8 +/- 3.9 fmol/10(6) cells) whereas there were significantly lower nuclear binding sites in perirenal adipocytes (16.5 +/- 5.5 fmol/10(6) cells) (P less than 0.05) and subcutaneous adipocytes (4.8 +/- 1.5 fmol/10(6) cells) (P less than 0.01). The apparent affinity in the three fat depots were similar with Kd values about 4 nM. The nuclear corticosteroid receptor in adipocytes was steroid specific, as neither unlabelled estradiol nor testosterone were able to displace the [3H]triamcinolone binding at concentrations up to 100 microM. However, unlabelled progesterone and promegestrone (R5020) were able to compete with triamcinolone-binding (by 50-80%). In order to investigate whether the nuclear corticosteroid binding in adipocytes were under influence of other hormones we examined the effects of lipolytic and antilipolytic compounds on the binding. Preincubation with isoproterenol and dibutryl-cAMP for 1 h was able to decrease the corticosteroid binding by 30-50%. However, the antilipolytic hormone insulin had no effect in preincubations performed for up to 2 h. In conclusion, high affinity nuclear corticosteroid receptors were found in rat adipocytes. These receptors exhibited regional variations and were modulated by lipolytic hormones.
Pedersen
S B
SB
University Clinic of Endocrinology and Internal Medicine, Aarhus Amtssygehus, Denmark.
Børglum
J D
JD
Møller-Pedersen
T
T
Richelsen
B
B
D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002467:Cell Nucleus / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism; D007700:Kinetics; D008297:Male; D011073:Polyamines / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013256:Steroids / Q000378:metabolism; D014221:Triamcinolone / Q000378:metabolism
D007328:Insulin; D011073:Polyamines; D011965:Receptors, Glucocorticoid; D013256:Steroids; D014221:Triamcinolone