Cytosolic and nuclear estrogen receptors in the isolated perfused rat liver.
The isolated perfused rat liver was investigated as a potential model to analyze binding of 17 beta-[3H]estradiol (E2) to cytosolic and nuclear estrogen receptors. Viability of the isolated perfused liver was monitored by measuring leakage of cytosolic enzymes into the perfusate. These studies indicated that the liver remained viable for at least a 90-min perfusion period although significant decreases in cytosolic estrogen receptor concentrations occurred during this perfusion period. Estrogen receptor loss was minimized by supplementing the red blood cell-free perfusion medium with 5 micrograms of insulin per ml. Uptake of [3H]E2 by hepatic cytosolic estrogen receptors of the isolated perfused liver was rapid as measured by partial purification of radiolabeled ligand receptor complexes after varying times of perfusion. Peak liver concentrations of receptor-bound E2 were achieved 15 min after the onset of perfusion when using livers from either male or female rats. After 15 min, radiolabeled cytosolic ligand receptor complexes decreased rapidly, reaching lowest concentrations at 60 min. Radiolabeled salt-extractable nuclear-binding sites increased up to 30 min and then decreased slightly between 30 and 90 min. Both 4S and 5S forms of nuclear binding sites were detected in the isolated perfused livers as evaluated by sedimentation analysis on 5 to 20% sucrose density gradients. Concentrations of radiolabeled cytosolic and nuclear receptors were greater in females than males at all perfusion periods examined when the initial concentration of [3H]E2 was 4 nm. Sex differences in receptor uptake were not as great when higher concentrations of [3H]E2 were added to the perfusion medium. These studies suggest that the isolated perfused liver is a suitable model to investigate short-term uptake of estrogens by cytosolic and nuclear receptors.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D003600:Cytosol / Q000032:analysis*; D004958:Estradiol / Q000378:metabolism; D005260:Female; D007016:Hypophysectomy; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000032:analysis*; D008297:Male; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains; D011960:Receptors, Estrogen / Q000032:analysis* / Q000378:metabolism; D012737:Sex Factors
D007328:Insulin; D011960:Receptors, Estrogen; D004958:Estradiol