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[Effect of testosterone on spermatogenesis: dynamics of testosterone secretion in adult rat testis].

OBJECTIVE: To study the effect of testosterone on spermatogenesis. METHODS: Testosterone concentrations were measured in testicular interstitial fluid (IF), and serum sample from the testicular artery, testicular veins on the surface of the testis, and the peripheral venous, inferior vena cave and proximal spermatic veins in adult SD rats. The left and bilateral veins at the proximal end of the spermatic cord were ligated respectively. Testosterone concentrations and weight of the testis were measured, and the changes of the testicular morphology were studied 3 days and 21 days after ligation respectively. RESULTS: The testosterone concentrations were highest in IF. The serum testosterone concentrations were highest in the testicular veins on the surface of the testis. The serum testosterone concentrations in the proximal spermatic veins, the testicular artery, the peripheral venous and inferior vena cave were (42.503 +/- 12.749), (42.503 +/- 12.749), (5.598 +/- 3.649), (2.533 +/- 1.719) and (2.418 +/- 1.495) mg/L respectively. Three days after the proximal spermatic veins were ligated bilaterally, the weights of the testis and the serum testosterone concentrations declined markedly. The epithelium of the seminiferous tubules degenerated slightly and part of the structure was indistinct. Three days after the left proximal spermatic veins were ligated, the weights of the left testis, the testosterone concentrations in the left IF and in the serum of the left testicular artery declined distinctly except the serum testosterone concentration of the peripheral venous, inferior vena cave and those of the right testis. The epithelium of the seminiferous tubules in the left testis degenerated slightly and part of the structure was indistinct. It was restored 21 days after ligation. CONCLUSIONS: A "small circulation" (testis-the testicular veins-the spermatic veins-the spermatic artery-the testicular artery-testis) may exist. The results indicate that after ligation of the proximal spermatic veins, the reduction of the testosterone concentrations and changes of testicular morphology is temporary and may be recovered.

Animals↗

[Trimethylsilylation of testosterone, epi-testosterone and determination of their ratio in urine by GC-MS].

The trimethylsilylation of testosterone and epitestosterone was discussed in detail in this report. Both derivative conditions under which testosterone and epi-testosterone were prepared into TMS-derivatives in the presence of mercaptoethanol as an antioxidizing agent and method for the analysis of the ratio of testosterone to epi-testosterone in urine, based on GC-MS, had been established. The conditions of detection were: carrier gas was helium, derivatives were separated with SE-54 fused silica capillary column, using temperature program and detected by using multiple ion detection mode in which the ion of m/z 432 was the monitoring ion. The method is rapid, sensitive and specific. For the ratio of testosterone to epi-testosterone (testosterone: 20 ng/microliters), there is a linearity between ratio 1:1 and 10:1 (r = 0.998), the limit of detection for testosterone and epi-testosterone is 1 ng, and the minimum concentration of detection in urine is 8 ng/ml.

Antioxidants↗

Oral testosterone undecanoate (Andriol) supplement therapy improves the quality of life for men with testosterone deficiency.

In a single-blind, placebo-controlled study, the effects of a 3-month oral administration of 160 mg/day testosterone undecanoate (Andriol) on the quality of life of men with testosterone deficiency were evaluated. The subjects included ten men with primary hypogonadism and 29 with andropause with sexual dysfunction as the most common problem. The changes in subjective symptoms were evaluated by the PNUH QoL scoring system and the St. Louis University Questionnaire for androgen deficiency in aging males (ADAM). Digital rectal examination (DRE) was performed and serum testosterone, prostate-specific antigen (PSA) and liver profile were monitored. Testosterone undecanoate treatment (n = 33) significantly improved sexual dysfunction and symptom scores of metabolic, cardiopulmonary, musculoskeletal and gastrointestinal functions compared to baseline and to placebo (n = 6). ADAM score also significantly improved after 3 months of treatment. Serum testosterone was significantly increased compared to pretreatment levels only in the testosterone undecanoate group. In the placebo group, no significant changes compared to baseline were found for testosterone levels and QoL questionnaires. No abnormal findings were detected on DRE or laboratory findings in either group. Adverse events, such as gastrointestinal problems and fatigue, were mild and self-limiting. It is concluded that androgen supplement therapy with oral testosterone undecanoate (Andriol) restores the quality of life through improvement of general body functions in men with testosterone deficiency.

Administration, Oral↗

Daily testosterone and gonadotropin levels are similar in azoospermic and nonazoospermic normal men administered weekly testosterone: implications for male contraceptive development.

Weekly intramuscular administration of testosterone esters such as testosterone enanthate (TE) suppresses gonadotropins and spermatogenesis and has been studied as a male contraceptive. For unknown reasons, however, some men fail to achieve azoospermia with such regimens. We hypothesized that either 1) daily circulating serum fluoroimmunoreactive gonadotropins were higher or testosterone levels were lower during the weekly injection interval, or 2) monthly circulating bioactive gonadotropin levels were higher in nonazoospermic men. We therefore analyzed daily testosterone and fluoroimmunoreactive gonadotropin levels as well as pooled monthly bioactive and fluoroimmunoreactive gonadotropin levels in normal men receiving chronic TE injections and correlated these levels with sperm production. After a 3-month control period, 51 normal men were randomly assigned to receive intramuscular TE at 25 mg (n = 10), 50 mg (n = 9), 100 mg (n = 10), 300 mg (n = 10), or placebo (n = 12) weekly for 6 months. After 5 months of testosterone administration, morning testosterone and fluoroimmunoreactive follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were measured daily for a 1-week period between TE injections. In addition, fluoroimmunoreactive and bioactive FSH and LH levels were measured in pooled monthly blood samples drawn just before the next TE injection. In the 100-mg and 300-mg TE groups, mean monthly fluoroimmunoreactive FSH and LH levels were suppressed by 86%-97%, bioactive FSH and LH levels by 62%-80%, and roughly half the subjects became azoospermic. In the 1-week period of month 6, daily testosterone levels between TE injections were within the normal range in men receiving placebo, or 25 or 50 mg of weekly TE, but were significantly elevated in men receiving 100 or 300 mg of weekly TE. At no point during treatment, however, were there significant differences in daily testosterone or fluoroimmunoreactive gonadotropin levels, or monthly bioactive gonadotropin levels between men achieving azoospermia and those with persistent spermatogenesis. This study, therefore, demonstrates that neither monthly nor daily differences in serum testosterone, or fluoroimmunoreactive or bioactive gonadotropins explain why some men fail to completely suppress their sperm counts to zero with weekly TE administration. Innate differences in the testicle's ability to maintain spermatogenesis in a low-gonadotropin environment may explain persistent spermatogenesis in some men treated with androgen-based contraceptive regimens.

Adult↗

Direct radioimmunoassay (RIA) of salivary testosterone: correlation with free and total serum testosterone.

Simple and sensitive direct RIA for determination of salivary testosterone was developed by using RSL NOSOLVEX TM (125 1) kit produced by Radioassay System Laboratories (Carson, California). In addition, a relationship between salivary and serum free and total testosterone concentrations was studied in randomly selected 45 healthy subjects, 5 females on oral contraceptive pills and 28 hypertensive patients on various treatment regimens. The lowest weight of testosterone detectable by our modified method was equivalent to 1 pg/ml of saliva, taking into account analytical variability. Intra- and interassay coefficients of variation were 5.09 +/- 2.7% and 8.2 +/- 5.9% respectively. Statistically significant correlations were found between salivary and serum free testosterone (r = 0.97) and salivary and serum total testosterone concentrations (r = 0.70-0.87). The exception to this was a group of hypertensive females in which no correlation (r = 0.14) between salivary and total serum testosterone was found. It is also of interest that, while salivary testosterone was significantly increased in subjects taking oral contraceptives and most of the hypertensive patients the total serum testosterone concentration was in normal range. Our findings suggest that determination of salivary testosterone is a reliable method to detect changes in the concentration of available biologically active hormone in the circulation.

Adult↗

Plasma androgen levels in men after oral administration of testosterone or testosterone undecanoate.

Plasma testosterone and androstenedione levels in men were measured after oral administration of free testosterone and testosterone undecanoate. Both androgens were determined by simultaneous, specific radioimmunoassays after separation and isolation by thin layer chromatography. While free unesterified testosterone had no effect on plasma androgen levels, a striking increase of both testosterone and androstenedione levels was noted after administration of testosterone undecanoate, which is otherwise only achieved by parenteral testosterone application. This effect of testosterone undecanoate is probably due to absorption via the lymph rather than via the portal vessels so that peripheral circulation is reached before metabolism in the liver. Testosterone undecanoate promises to be an effective medication for oral androgen replacement.

Adolescent↗

Relation between circulating levels of testosterone lh and fsh in intact and castrated, adult, male rats after testosterone administration.

Serum levels of LH, FSH and testosterone were measured by radioimmunoassay in intact and castrated, adult, male rats after testosterone was administered subcutaneously for seven days in doses ranging from 25 to 200 mug per 100 g body weight per day. Such treatment increased circulating testosterone both in intact and castrated rats, but its effects on serum gonadotrophins were different in these animal groups. All doses of testosterone suppressed serum LH and FSH in the normal rat. In the castrates, treatment with the lowest dose of testosterone resulted in serum LH levels significantly above the high castrate levels, while serum FSH tended to drop. Administration of the highest doses of testosterone did not depress serum LH and FSH in the castrates to those of intact, normal animals, though serum testosterone in these castrates was much higher than in normal, male rats. It is concluded, that the sensitivity of the hypothalamic-pituitary system for daily, subcutaneous testosterone administration during seven days is not the same in the intact and castrated, adult, male rat and that testicular factors different from testosterone may play a role in regulating production and/or secretion of gonadotrophins by the hypophysis in male animals.

Animals↗

Hydroxylation of testosterone in the human testis. Identification of 4-androstene, 7 alpha,17 beta-diol-3-one (7 alpha-hydroxytestosterone) as a metabolite of testosterone.

Homogenates of normal or cryptorchid, human testes were incubated with [3H]testosterone and a NADPH-generating system. [3H]4-andostene,7 alpha,17 beta-diol-3-one (7 alpha-OH-testosterone) was isolated and identified from such incubations. To our knowledge this is the first demonstration that 7 alpha-OH-testosterone is a metabolite of testosterone in the human testis. 4-Andro-stene,6 beta,17 beta-diol-3-one (6 beta-OH-testosterone) and 4-androstene,16 alpha,17 beta-diol-3-one (16 alpha-OH-testosterone) were also identified as testosterone metabolites. The specific activity of both testosterone 6 beta- and 16 alpha-hydroxylase was higher than that of 7 alpha-hydroxylase. Pre-pubertal or cryptorchid human testis tissues seem in our study to have higher testosterone 6 beta- and 7 alpha-hydroxylase activity than normal adult testis tissue.

Adolescent↗

New injectable testosterone ester maintains serum testosterone of castrated monkeys in the normal range for four months.

Two groups of four long-term orchidectomized cynomolgus monkeys, Macaca fascicularis, weighing 2.8-4.6 kg received either a single intramuscular injection of 40 mg of a new testosterone ester, testosterone-trans-4-n-butylcyclohexyl-carboxylate (20 Aet-1) in an aqueous suspension or 32.8 mg testosterone oenanthate dissolved in sesame oil. Both preparations contained equal amounts of testosterone, namely 23.6 mg. Testosterone oenenthate injections resulted in supraphysiological serum testosterone levels for eight days followed by a rapid decline so that the lower physiological limit was reached after three weeks. In contrast, 20 Aet-1 produced a moderate increase of serum testosterone levels into the physiological range. Serum testosterone remained in this range for a period of 18 weeks. Thus it appears that the 20 Aet-1 may provide a long desired, new modality of testosterone substitution for hypogonadal men as well as for methods of male fertility control.

Animals↗