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Androgens enhance in vivo 2-deoxyglucose uptake by rat striated muscle.

Testosterone propionate causes a striking increase in in vivo uptake of 2-deoxyglucose by the levator ani muscle of immature male rats. Autoradiography showed the labeled 2-deoxyglucose to be uniformly distributed over the entire muscle. Liquid scintillation counting of whole muscles allowed quantification of 2-deoxyglucose uptake. After a single sc injection of testosterone propionate, no enhancement of 2-deoxyglucose uptake could be seen before 3.5 h, at which time uptake was increased 2-fold; maximum enhancement (4-fold) was attained at 12 h. 2-Deoxyglucose uptake remained elevated at twice the control value at 72 h. Muscle weight did not increase until sometime after 24 h; by 39 h, it was 50% greater than the control value; by 72 h, wet weight was double the control value. The effect of testosterone propionate probably is mediated by specific androgen receptors, since the administration of either 5 alpha-dihydrotestosterone or fluoxymesterone, potent androgens, caused severalfold increases in 2-deoxyglucose uptake in the levator ani muscle, while administration of 17 beta-estradiol, corticosterone, and etiocholanolone, an inactive metabolite of testosterone, did not. Furthermore, the effect of testosterone propionate was blocked by simultaneous administration of an androgen antagonist, cyproterone acetate. Testosterone propionate also enhanced uptake of 2-deoxyglucose in the bulbocavernosus (253% over control) and extensor digitorum longus muscles (150% over control), but not in the biceps brachii or soleus. Increased glucose uptake may be an important early step in the anabolic response of muscle to androgens.

Androgens↗

Gonadal steroid preservation of central synaptic input to hamster facial motoneurons following peripheral axotomy.

In recent work, we have demonstrated that testosterone propionate accelerates recovery from facial nerve injury in the adult male hamster. Central synaptic stripping following peripheral motor neuron damage is a well-established component of the injury response. Gonadal steroids regulate synaptogenesis in the normal nervous system. In this study, we tested the hypothesis that testosterone propionate administration at the time of facial nerve transection alters the synaptic connectivity of injured facial motoneurons. Adult hamsters were subjected to right facial nerve transection at the level of the stylomastoid foramen. Half the animals received subcutaneous implants of testosterone propionate; the other half were sham implanted. At 5 days postoperative, the animals were killed by intracardiac perfusion-fixation, and the control and axotomized facial nuclear groups from the brainstems of nonhormone- and testosterone propionate-treated animals processed for routine transmission electron microscopy. Quantiative analysis of the synaptic ratio (percent somal membrane covered by synaptic profiles) and the average length of axosomatic synapses was accomplished. The results indicate that axotomy alone resulted in an 81% reduction in the synaptic ratio and a 26% decrease in the average synaptic length of axosomatic synapses. Exposure to testosterone propionate from the time of facial nerve transection resulted in only a 48% reduction in the synaptic ratio and a 16% decrease in the average synaptic length of axosomatic synapses following injury. Thus, testosterone propionate significantly attenuated the amount of synaptic stripping that occurred at 5 days postoperative and the decrease in average length of the remaining synapses as well. It is concluded that gonadal steroids modulate central synaptic plasticity following peripheral nerve injury. The results are discussed in light of our recent findings of steroidal effects on the central astrocyctic response to facial nerve injury as well.

Animals↗

The regulation by thyroid hormones and androgen of epidermal growth factor synthesis in the submandibular gland and its plasma concentrations in mice.

The effects of androgen and thyroid hormones on epidermal growth factor (EGF) synthesis in the submandibular gland and on plasma EGF concentrations in mice were examined. Testosterone propionate was administered alone or in combination with L-thyroxine (T4) to female mice for 2 weeks. The submandibular EGF concentrations were increased by the administration of testosterone propionate in a dose-dependent fashion; the maximal increase, 20-fold, being produced by a dose of 2 mg every other day. The EGF levels were increased sevenfold by T4, which also enhanced the stimulatory effect of suboptimal doses of testosterone propionate. Cyproterone acetate, an anti-androgen, inhibited the testosterone propionate-induced increase, but not the T4-induced increase. Plasma EGF concentrations were raised by testosterone propionate but not by T4. Both hormones stimulated the accumulation of 4.7 kb preproEGF mRNA in the submandibular gland, which occurred almost in a parallel manner with the increase in submandibular EGF concentrations. These results suggest that EGF synthesis in the submandibular gland is regulated by alterations in the level of its mRNA by thyroid hormones and androgen, and that the rise in plasma EGF concentrations is under the influence of androgen but not of thyroid hormones.

Animals↗

Effects of testosterone, testosterone metabolites and anti-androgens on the function of the male accessory glands in the rabbit and rat.

The androgenic potencies of testosterone, 5alpha-dihydrotestosterone, 5alpha-androstane-3alpha,17beta-diol and 5alpha-androstane-3beta,17beta-diol towards the prostate, glandula seminalis+glandula vesicularis, ampullae and epididymis were evaluated after administration to castrated rabbits. The influence of cyproterone acetate, stilboestrol and medrogestone on accessory gland function was also investigated in rabbits and rats. In the rabbit it was found that the minimum dose of testosterone propionate that would maintain the function of all accessroy glands at normal levels was approximately 200 microgram/animal/day. Higher levels of testosterone propionate overstimulated the function of the prostate, glandula seminalis+glandula vesicularis and ampullae, but did not affect the epidiymis. Whereas testosterone propionate and 5alpha-dihydrotestosterone propionate were essentially equipotent in their capcity to support growth and secretory activity and stimulated all the accessory glands, 5alpha-androstane-3alpha,17beta-diol dipropionate had a pronounced differential effect; it was considerably more potent than testosterone propionate in promoting secretion in the prostate, but was ineffective in maintaining the function of the epididymis. 5alpha-Androstane-3beta,17beta-diol dipropionate was the weakest androgen tested. Evidence also indicated that the potency of a steroid can depend on whether it is administered as its free or esterified form. Cyproterone acetate suppressed fructose secretion in the prostate of the rabbit but had no adverse effects on the function of the epididymis in either the rabbit or rat. Stilboestrol was the most potent anti-androgen tested and medrogestone the least effective.

Androgen Antagonists↗

Androgen-dependent fighting behaviour in male mice.

The influence of testosterone propionate, 17 alpha-methyl-testosterone and cyproterone acetate on isolation induced fighting behaviour of mice was studied in a simple testing procedure. Decreased aggressiveness has been established in mature, sexual experienced and isolated male mice both following castration and administration of the antiandrogen cyproterone acetate, respectively. Replacement therapy with testosterone propionate s.c. and 17 alpha-methyl-testosterone p.o. has been shown to restore the decreased level of aggresiveness after castration.

Aggression↗

Neonatal androgen effects on conditioned emotional response extinction of adult rats under various hormonal regimens.

Intact male rats and spayed female rats received 0.5 mg testosterone propionate in the first 24 hours after birth. They were divided into different groups, administered peanut oil, progesterone, oestradiol benzoate and testosterone propionate in adulthood. The animals were conditioned to fear with a shock paired sound. The experimental test consisted of investigating the conditioned emotional response (CER) extinction. It was verified that as to conditioned emotional response extinction, neonatally androgenized males were different from control males; neonatally androgenized females, testosterone propionate treated in adulthood were also different from control females. There was no significant difference between neonatally androgenized females oestradiol benzoate treated in adulthood and their control group. In comparing neonatally androgenized females having different hormonal treatment in adulthood, we verified that testosterone propionate treated females were significantly different from those treated with progesterone. On the other hand, neonatally androgenized females treated with oestradiol benzoate in adulthood were different from those that have had the same hormonal manipulation in neonatal period, but that were injected with progesterone in adulthood. It was concluded that conditioned emotional response extinction is facilitated by neonatal androgenization in non-oestrogenized male and female rats.

Age Factors↗

Effect of hormonal steroids on lactate dehydrogenase activity and its isozymes in the coagulating gland of castrated male rats.

The effect of castration and treatment with testosterone propionate, androstenedione and oestradiol-17 beta on the coagulating gland LDH activity and its isozymes were studied. Castration reduced the total LDH activity and administration of testosterone propionate alone was able to restore it back to sham operated control levels. Regarding LDH isozymes, the LDH3 and LDH4 responded very similarly to testosterone and oestradiol. LDH5 was significantly increased only in the androgen treated groups. The post-castration decrease in total M-subunits was restored to sham operated control levels in the testosterone propionate treated group. This was mainly contributed by the LDH3, LDH4 and LDH5 isozymes. The changes observed due to testosterone propionate and androstenedione treatments have been attributed to the rejuvenation of tissue from the castration induced losses. The oestradiol induced events have been associated with the growth of fibro muscular layers observed under similar experimental conditions.

Androstenedione↗

[Effect of testosterone on carboxypeptidase H activity in the murine hypothalamus and hypophysis].

Effects of single intraperitoneal administration of testosterone propionate (3 or 30 mg/kg body weight) on activity of carboxypeptidase H in pituitary body and hypothalamus of female white mouse were studied. It was found that testosterone propionate treatment (3 and 30 mg/kg body weight) increased the carboxypeptidase H activity in pituitary through 0.5 hour and it decreased one in 24 hours after treatment. The carboxypeptidase H activity in hypothalamus was lower as compared with control animals in 24 hours after testosterone propionate treatment in the dose 3 mg/kg body weight. However, the carboxypeptidase H activity in hypothalamus was lower in 0.5 and 24 hours and it was higher in 4 h after testosterone propionate treatment in the dose 30 mg/kg body weight as compared with the control. These data suggest that testosterone affects the carboxypeptidase H activity by changing the level of enzyme gene expression.

Animals↗

Steroid feedback on luteinizing hormone secretion during sexual maturation in the pig.

The effects of gonadal secretions on the release of LH and the stimulation of LH secretion by oestradiol have been investigated in newborn male and female miniature pigs; the differences in the feedback action of testosterone in newborn and pubertal male pigs were also studied. Hemi-orchidectomy or orchidectomy of 1-week-old pigs had no effect on the level of LH in the plasma; total orchidectomy significantly reduced the levels of testosterone (P less than 0.01) and progesterone (P less than 0.05). In female pigs ovariectomized at 1 week of age, the concentration of LH in the plasma decreased, with a strong negative correlation between the level of LH and age (r = -0.41; P less than 0.05). The plasma concentration of progesterone was generally low and unaffected by ovariectomy. Orchidectomy and treatment of male pigs, at 1 week of age, with testosterone (6 mg/kg body weight) had no effect on the plasma concentration of testosterone 24 h after treatment. If testosterone propionate was given rather than testosterone, the level of LH was significantly reduced (P less than 0.001) 24 h after the injection and the concentration of testosterone in the plasma corresponded to that found in the intact adult male pig. Treatment with oestradiol or oestradiol benzoate did not affect the concentration of LH. Orchidectomy and treatment of pubertal male pigs with testosterone propionate resulted in a significantly (P less than 0.001) higher concentration of testosterone in the plasma, compared with newborn pigs treated similarly, but the level of LH was unchanged. This suggests that there is a more rapid rate of clearance of testosterone in the newborn than in the pubertal male miniature pig and that the negative feedback of testosterone is not mediated by aromatization in the newborn animal and it declines before or during puberty. Treatment of newborn intact male and female and gonadectomized male pigs with oestradiol benzoate produced similar variations in the plasma level of oestradiol in all groups of animals. In the female pigs, however, a surge-like release of LH was observed 60--72 h after the injection of oestradiol benzoate, suggesting that the stimulatory feedback mechanism can operate soon after birth and that the response is sexually dimorphic.

Animals↗

Growth-stimulative effect of estrogen on androgen-dependent Shionogi carcinoma 115.

The stimulative effect of 17 beta-estradiol on the growth of androgen-dependent Shionogi carcinoma 115 and estrogen receptor in the tumor were studied. The incorporation of 17 beta-[3H]estradiol following a single injection of 17 beta-[3H]estradiol into tumor-bearing animals was 5- to 20-fold higher in the tumor than in the spleen and blood. Scatchard plot analyses showed that the tumor cytosol possessed a 17 beta-estradiol-binding site having a high affinity for 17 beta-estradiol [Kd 1.1 +/- 0.1 nM (S.E.)]. Competition experiments demonstrated that the 17 beta-estradiol binding was specific only for estrogenic compounds. Using sedimentation coefficient obtained by high-salt sucrose gradient (4.0S) and Stokes radius obtained by gel chromatography on Sephadex G-200 (46 A), the molecular weight of 17 beta-estradiol-binding component in the tumor cytosol was estimated to be 76,400. In castrated DS mice, a slight but significant increase in growth of s.c. grafted tumors was found by daily s.c. injections of either 17 beta-estradiol or 10 micrograms per mouse of testosterone propionate. Growth of the tumor maintained by 10 micrograms of testosterone propionate was augmented markedly by the addition of 4 micrograms of 17 beta-estradiol, and the growth approached the level induced by 100 micrograms of testosterone propionate. Simultaneous injections of bromocryptine inhibited an increase in 17 beta-estradiol-induced prolactin secretion but had no effect on the 17 beta-estradiol-enhanced tumor growth. These results demonstrate for the first time the stimulative effect of estrogen on the growth of androgen-dependent Shionogi carcinoma 115. The tumor contains typical estrogen receptor, which might be able to transmit estrogen signal to tumor cell nuclei with regard to tumor growth.

Androgens↗

Androgen-estrogen synergy in rat levator ani muscle: glucose-6-phosphate dehydrogenase.

The effects of castration and hormone administration on the activity of glucose-6-phosphate dehydrogenase in the rat levator ani muscle were studied. Castration caused a decrease in enzyme activity and in wet weight of the levator ani muscle. Chronic administration of testosterone propionate increased glucose-6-phosphate dehydrogenase activity in the levator ani muscle of castrated rats; the magnitude of the recovery of enzyme activity was related to the length of time of exposure to testosterone propionate after castration as well as to the length of time the animals were castrated. The longer the period of castration before exposure to testosterone propionate, the greater the effect. This result may be related to previously reported castration-mediated increases in androgen receptor binding in muscle. Dihydrotestosterone was less effective than testosterone propionate in enhancing glucose-6-phosphate dehydrogenase activity in the levator ani muscle from castrated rats; estradiol-17 beta alone was ineffective. Combined treatment with estradiol-17 beta and dihydrotestosterone, however, was as effective as testosterone alone. Thus, androgens and estrogens may exert synergistic effects on levator ani muscle.

Androgens↗

Reversal by testosterone of atrophy of accessory genital glands of castrated male sheep. A histologic and morphometric study.

The histologic features of male accessory genital glands of entire sheep (group I), castrated sheep (group II), castrated sheep treated with 40 daily intramuscular injections of 50 milligrams testosterone propionate (group III), and castrated sheep treated with 600 milligrams testosterone propionate 72 hours before death (group IV) were compared. Sheep were castrated at 3 months old and all sheep were killed when 15 months old. Volume fractions of glandular tissue, intralobular fibromuscular tissue and perilobular fibromuscular tissue of the seminal vesicles and Cowper's glands fluctuated significantly (P less than 0.05) during postcastration atrophy and after repeated testosterone treatment. Atrophy in sheep in group II was least in the prostate but greatest in Cowper's glands, seminal vesicles and ampullae of vasa deferentia. Seminal vesicle plexi, whose cytons had a statistically significant (P less than 0.05) degree of shrinkage, also were atrophied. After treatment with testosterone the postcastration atrophy of plexal neurons was almost reversed in sheep in group III. There also was hypertrophy of epithelial cells but the testosterone treatment failed to reduce to normal the fibromuscular volume fraction of the accessory genital glands. Testosterone propionate treatment of sheep in group IV failed to elicit appreciable morphologic changes. These results are compared with our previous findings on the content and uptake of zinc by the accessory genital glands. It is suggested that accumulation of zinc in the accessory genital glands of sheep is not necessarily closely linked to normal histologic appearance.

Animals↗

Influence of pituitary and adrenocortical hormones on prostatic secretion protein, a major protein in rat prostate.

Prostatic secretion protein (PSP) is an androgen-sensitive, quantitatively important protein in rat ventral prostate which has been shown to inhibit the nuclear uptake and decrease the DNA-binding capacity of the androgen-receptor complex. In the present study, the influence of the pituitary, adrenals, and gonads on the concentration of PSP in the prostate was studied. Hypophysectomy decreased the concentration of PSP to about 10% of the control level, an effect similar to that obtained by castration. No effects on prostatic wet weight or PSP concentration were observed following substitution of hypophysectomized rats with human growth hormone, rat prolactin, or rat growth hormone. On the other hand, PSP concentration as well as wet weight of the prostate were normalized in hypophysectomized rats after administration of testosterone propionate. These results are in line with a direct extrahypophyseal effect of androgens on the prostate. Adrenalectomy did not affect the concentration of PSP, nor the wet weight of the prostate. Administration of estramustine decreased the wet weight of the prostate but did not affect the prostatic concentration of PSP in normal rats. Combined treatment with testosterone propionate and estramustine seemed to increase both the wet weight and the concentration of PSP more than administration of testosterone propionate alone. These results indicate a synergistic effect between estramustine and testosterone propionate.

Adrenal Cortex Hormones↗

Altered testosterone feedback in pubertal male rats raised on reduced caloric intake.

A study was conducted to explore the hypothesis that reduced caloric intake increases the negative feedback efficacy of testosterone on gonadotropin secretion. Daily subcutaneous injections of testosterone propionate in oil were administered to either age-matched (37-day-old) or weight-matched (mean = 178 g) ad libitum-fed (control) or underfed rats that received 60% of normal daily food intake from the age of 20 days. Daily doses of testosterone propionate ranged from 25 to 400 micrograms/100 g body weight. Results indicate that reduced caloric intake increases the suppression of serum LH by testosterone propionate in both age-matched and weight-matched underfed rats. On the other hand, the response of serum FSH concentrations to testosterone propionate was affected by reduced diet only in the weight-matched animals. Results suggest that the increased negative feedback of testosterone on LH secretion is a specific diet-induced effect and not merely the result of reduced growth. The altered response in FSH secretion, however, may represent age-associated changes in sensitivity to testosterone rather than diet-induced effects.

Animals↗

Testosterone effects on mares during synchronization with altrenogest: FSH, LH, estrous duration and pregnancy rate.

Twelve mares fed altrenogest for 14 d were used to study the effects of a single injection of testosterone propionate on concentrations of follicle-stimulating hormone (FSH) during diestrus, and to relate the normal and perturbed patterns of FSH secretion to subsequent estrous characteristics and fertility. Seven of 12 mares received testosterone propionate at 200 micrograms/kg of body weight on d 5 of progestogen feeding. Mares were teased and blood samples were drawn daily; all mares were artificially inseminated at the first estrus after progestogen treatment. Testosterone propionate treatment caused a 50% reduction in concentration of FSH in plasma within 24 h, and this effect persisted through 48 h after injection. Within 4 d after the suppression of FSH secretion, concentrations of FSH rebounded and were significantly elevated compared with control values during the last 4 d of progestogen feeding. Testosterone propionate at this dosage also elicited estrous behavior in five of seven treated mares within 24 h after injection. After withdrawal of progestogen feeding, the interval to onset of estrus, duration of estrus, and magnitude of the luteinizing hormone peak were similar between groups (P greater than .05). Six of seven treated mares and three of five control mares became pregnant to breeding on the first estrus after progestogen treatment. Because manipulation of the normal pattern of FSH secretion during diestrus did not affect estrous characteristics or fertility of the subsequent estrus, such treatment may have potential as a means of synchronizing FSH surges during diestrus in the mare.

Animals↗

Effects of testosterone, dihydrotestosterone and estradiol on gonadotropin release after gonadotropin releasing hormone administration in cyclic mares.

Sixteen intact cyclic mares were treated on the fourth day of estrus and then every other day for a total of six injections with 1) testosterone propionate, 2) dihydrotestosterone (DHT) benzoate, 3) estradiol (E2) benzoate or 4) safflower oil. Mares were given gonadotropin releasing hormone (GnRH) on Day 3 of estrus (pretreatment) and again 24 h after the last steroid or oil injection. Treatment with testosterone propionate resulted in a greater (P less than 0.05) follicle-stimulating hormone (FSH) response to the second injection of GnRH compared with all other treatments. Treatment with DHT benzoate also resulted in greater (P less than 0.05) FSH response to GnRH compared with control and E2 benzoate-treated mares. Testosterone propionate and E2 benzoate administration suppressed (P less than 0.05) the normal diestrous rise in FSH concentrations exhibited by the control and DHT benzoate-treated mares. Steroid treatment did not affect the luteinizing hormone (LH) response to GnRH, although testosterone propionate treatment did suppress concentrations of LH in daily blood samples during Days 3 to 6 of treatment. It is concluded that testosterone's effect on FSH after GnRH treatment observed in this and previous experiments can be attributed to two different properties of the hormone or its metabolites acting simultaneously. That is, testosterone increased the secretion of FSH in response to GnRH as did DHT (an androgenic effect). At the same time, testosterone suppressed FSH concentrations in daily blood samples in a manner identical to that of E2 benzoate (an estrogenic effect).

Animals↗

Counteraction of gonadal steroid inhibition of luteinizing hormone release by naloxone.

The effect of naloxone on the negative feedback action of gonadal steroids on luteinizing hormone (LH) release was studied in castrated male and female rats. The reduction by estradiol benzoate or testosterone propionate of elevated serum LH levels in rats ovariectomized for 4 weeks was reversed by a single injection of naloxone. Injection of estradiol benzoate, together with progesterone, similarly reduced serum LH levels in ovariectomized rats, and this inhibition was partially reversed by naloxone. A single injection of testosterone propionate decreased serum LH levels in male rats castrated 48 h earlier, and naloxone completely blocked the androgen-induced inhibition of LH secretion. Chronic administration of either testosterone propionate or morphine sulfate to castrated male rats prevented the postcastration rise of serum LH. The decrease of hypothalamic LHRH in castrated rats was completely blocked by testosterone propionate or morphine sulfate administration. These results are believed to indicate that hypothalamic opiates are involved in gonadal steroid feedback inhibition of LH release.

Animals↗

Diethylstilbestrol potentiates and testosterone antagonizes the action of 3-methylcholanthrene on benzo(a)pyrene metabolism in Hep G2 cells.

We have used the human hepatoma cell line, Hep G2, to examine the ability of hormones and xenobiotics to modulate the hepatic induction of benzo(a)pyrene hydroxylase and epoxide hydrolase. Hep G2 cells were cultured in Eagle's Minimum Essential Medium supplemented with 10% fetal calf serum. 3-Methylcholanthrene, diethylstilbestrol, testosterone propionate, and combinations of 3-methylcholanthrene, and each of the hormones were added directly to the culture media. We subsequently studied the metabolism of benzo(a)pyrene using cell lysates of the Hep G2 cells. Metabolites were quantitated by high-performance liquid chromatography (HPLC) using fluorodetection. Exposure to 3-methylcholanthrene alone resulted in an eightfold increase in total benzo(a)pyrene metabolites with a change of the predominant metabolite from the 3-hydroxybenzo(a)pyrene to the carcinogenic pathway of the benzo(a)pyrene-7,8-diol. Diethylstilbestrol and testosterone propionate resulted in small, but significant, decreases in metabolism of benzo(a)pyrene. When exposed in combination with 3-methylcholanthrene, testosterone propionate antagonized and diethylstilbestrol potentiated the metabolism of benzo(a)pyrene. 3-Methylcholanthrene, diethylstilbestrol, and combinations of 3-methylcholanthrene and diethylstilbestrol or testosterone propionate resulted in increased epoxide hydrolase activity as compared to controls. These results, carried out in a human hepatoma cell line, lend support to a concern for potentiated toxicity and carcinogenicity following exposure to complex chemical mixtures.

Benzo(a)pyrene↗