T. T. Kurowski

Countereffects of compensatory overload and glucocorticoids in skeletal muscle: Androgen and glucocorticoid cytosol receptor binding

Male hypophysectomized rats were divided into a control group and a group that was subjected to compensatory overload of plantaris muscles for 7 days following surgical removal of synergistic musculature. These groups received daily subcutaneous injections of either cortisone acetate (CA) (100 mg/kg b.wt) or the vehicle for 7 days starting at the time the overload was initiated. Plantaris muscle weights were atrophied by 25% in CA-treated controls, were hypertrophied by 45% in the vehicle-treated overloaded group, and remained similar to vehicle-treated controls in the group receiving both treatments. Scatchard analyses of specific binding of [3H]methyltrienolone (R1881), a synthetic androgen that binds to androgen receptors, were non-linear in plantaris muscle cytosols of vehicle-treated control and overloaded groups and were resolved by a two-component binding model. The lower affinity component, which was attributed to binding of methyltrienolone to a glucocorticoid receptor, disappeared in glucocorticoid-treated rats as evidenced by linear Scatchard plots. Receptor concentrations of the androgenic component of [3H]methyltrienolone binding were unchanged by CA treatment and were significantly increased only in the vehicle-treated overloaded group. [3H]Dexamethasone cytosol binding was increased nearly 2-fold in plantaris muscles of vehicle-treated overloaded animals (64 +/- 3 fmol/mg protein) as compared to those in vehicle-treated controls (32 +/- 2 fmol/mg protein), but was decreased to 2 and 4 fmol/mg protein in the CA-treated controls and CA-treated overloaded groups respectively. These results show that overload and glucocorticoids have opposing actions skeletal muscle, but the overload in the presence of glucocorticoids did not stimulate an increase in androgen cytosol receptor binding.

Skeletal muscle cytosol [3H]methyltrienolone receptor binding and serum androgens: effects of hypertrophy and hormonal state.

Normal, castrated, and hypophysectomized male rats underwent compensatory hypertrophy of plantaris muscles following surgical removal of their synergistic gastrocnemius muscles. The increases in muscle wet weights above control values, determined when the muscles were in stable-state hypertrophy, were as follows: normal 50%, castrated 50%, and hypophysectomized 32%. There were marked differences in concentration of serum androgens between surgical groups, yet no increases in testosterone or 5 alpha-dihydrotestosterone were observed as a result of hypertrophy. The amount of testosterone binding to serum proteins (approx 94%) was reduced only in hypophysectomized animals that underwent muscle growth. Cytosol androgen receptor specific binding (fmol/mg protein), measured using saturating concentrations of [3H]methyltrienolone (R1881) at 4 degrees C for 20 h for exchange with endogenous steroid, was significantly increased in hypertrophied muscles of normal, (1.77 +/- 0.17 vs 1.16 +/- 0.21), castrated (2.27 +/- 0.20 vs 1.46 +/- 0.03) and hypophysectomized (6.23 +/- 0.56 vs 3.64 +/- 0.30) animals. Receptor dissociation constants (Kd) were approx 10(-10) M in all groups and were not altered by the hypertrophy. These findings show that a major adaptation to skeletal muscle enlargement is an augmentation of cytosol [3H]methyltrienolone receptor binding capacity. This effect occurs in a normal or androgen deficient state.

Depletion of [3H]methyltrienolone cytosol binding in glucocorticoid-induced muscle atrophy.

Abstract The present study was undertaken to determine cytosol binding properties of [3H]methyltrienolone, a synthetic androgen, in comparison with [3H]dexamethasone, a synthetic glucocorticoid, under conditions of glucocorticoid excess in skeletal muscle. Male hypophysectomized rats received either seven daily subcutaneous injections of cortisone acetate (CA) (100 mg·kg- 1 body wt) or the vehicle, 1% carboxymethyl cellulose. Following treatment, both [3H]dexamethasone and [3H]methyltrienolone-receptor concentrations were decreased from those in vehicle-treated rats by more than 90 and 80%, respectively, in CA-treated animals. Scatchard analysis of [3H]methyltrienolone binding in muscles of vehicle-treated animals became nonlinear at high concentrations of labeled ligand and were reanalyzed by a two-component binding model. The lower affinity, higher capacity component, which was attributed to binding of methyltrienolone to a “dexamethasone” component, disappeared in muscles of CA-treated rats and Scatchard plots were linear. Receptor concentrations of the higher affinity lower capacity “methyltrienolone” component were similar in muscles of vehicle-treated and CA-treated groups. From competition studies, the high relative specificities of glucocorticoids for [3H] methyltrienolone binding in muscles of vehicle-treated animals were markedly reduced by CA treatment. In addition, the binding specificity data also showed strong competition by progesterone and methyltrienolone for [3H]dexamethasone binding and estradiol-17β for [3H]methyltrienolone binding. These results demonstrate that most of the [3H]methyltrienolone binding is eliminated under in vivo conditions of glucocorticoid excess. Furthermore, the competitiveness of various steroids for receptor binding suggests that rat muscle may not contain classic (ligand-specific) glucocorticoid and androgen receptors.