E. Lacroix

Pre-exercise branched-chain amino acid administration increases endurance performance in rats

This study investigated the effects of pre-exercise branched-chain amino acid (BCAA) administration on blood ammonia levels and on time to exhaustion during treadmill exercise in rats. Adult female Wistar rats were trained on a motor driven treadmill. After a 24-h fast, rats were injected intraperitoneally (i.p.) with 1 mL of placebo or BCAA (30 mg), 5 min before performing 30 min of submaximal exercise (N = 18) or running to exhaustion (N = 12). In both cases, rats were sacrificed immediately following exercise, and blood was collected for the measurement of glucose, nonesterified fatty acid (NEFA), lactic acid, BCAA, ammonia, and free-tryptophan (free-TRP) levels. Control values were obtained from sedentary rats that were subjected to identical treatments and procedures (N = 30). Plasma BCAA levels increased threefold within 5 min after BCAA administration. Mean run time to exhaustion was significantly longer (P < 0.01) after BCAA administration (99 +/- 9 min) compared with placebo (76 +/- 4 min). During exercise, blood ammonia levels were significantly higher (P < 0.01) in the BCAA treated compared with those in the placebo treated rats both in the 30-min exercise bout (113 +/- 25 mumol.L-1 (BCAA) vs 89 +/- 16 mumol.L-1) and following exercise to exhaustion (186 +/- 44 mumol.L-1 (BCAA) vs 123 +/- 19 mumol.L-1). These data demonstrate that BCAA administration in rats results in enhanced endurance performance and an increase in blood ammonia during exercise.

Influence of age on the formation of 5α-androstanediol and 7α-hydroxy-testosterone by incubated rat testes

Abstract Testes from rats of different ages were incubated with or without tritiated testosterone. The exogenously-added or endogenously-produced testosterone is mainly metabolized to 7α-hydroxylated testosterone in adult animals, and to Set-reduced metabolites (especially 5α-androstanediol) in immature animals.

The biosynthesis of estrogens by cow follicles

Abstract Separated and recombined granulosa and thecal cells of ripening cow follicles were incubated in vitro in the presence of different steroid precursors involved in the Δ 4 and Δ 5 -pathways. The experiments indicate that both in the granulosa and the thecal cells the transformation of pregnenolone to androstenedione occurs predominantly through the Δ 5 -pathway. Although both cell types are able to transform androstenedione to estrogens, this capacity is very small in the thecal tissue as compared to the granulosa cells. Incubations of combined granulosa and thecal cells yielded larger amounts of estrogens than the incubations of each cell type separately, indicating a positive interaction between both cell types in the experimental conditions.

The conversion of testosterone to 7α-hydroxy-testosterone by incubated rat testes

Abstract Incubations of testes of adult rats with testosterone yield rather important amounts of a very polar metabolite which is identified as 7α-hydroxytestosterone. The identification of the metabolite is based on chromatography, spectrophotometry, fluorimetry, counter current distribution and NMR spectrometry.

The influence of gonadotrophin (HCG) treatment on the steroidogenesis by incubated rat testes

Abstract The production of steroids and the metabolism of testosterone by the incubated testes from normal and HCG treated rats were studied. The production experiments show that HCG (3 UI/day for 10 days) induces an increase of testosterone and 5α-androstanediol production and a strong depression of 7α-hydroxytestosterone formation. The metabolism experiments indicate an increased transformation of testosterone to 5-reduced metabolites and a decreased conversion to 7α-hydroxytestosterone by the testes of HCG treated rats. It is concluded that HCG provokes a shift in the testicular metabolism pattern of testosterone from the 7α-hydroxylation to the 5α-reduction pathway.

[4-14C]-Testosterone metabolism and steroid production by incubated whole testes, seminiferous tubules and interstitial tissue from rats

Abstract In incubated testes of young rats (30 days old), 5α-reduction predominates over 7α-hydroxylation. This 5α-reductase activity appears to be located predominantly in the interstitial tissue. In incubated testes of mature rats (120 days old) 7α-hydroxylation is more important than 5α-reduction. This 7αzydroxylation mainly occurs in the interstitial tissue, while 5α-reduction predominates in the seminiferous tubules. During long term treatment with HCG, 7α-hydroxylation in the incubation of whole testes and of interstitial tissue decreases to low levels and the steroid metabolism shifts to 5α-reduction.

Regulation of the pituitary 5α-reductase activity by gonadotropin releasing hormone and testosterone in the adult male rat

Intact or castrated adult male rats were treated for nine days with GnRH (10 micrograms/day), the synthetic GnRH goserelin (100 micrograms/day) or the GnRH-antagonist Org 30276 (250 or 500 micrograms/day). In some series, 1 mg testosterone propionate was administered alone, or in combination with goserelin or Org 30276. The in vitro metabolism of [1 alpha,2 alpha-3H]testosterone by pituitary and hypothalamic homogenates was investigated in combination with the estimation of plasma concentrations of testosterone and gonadotropins. No qualitative or quantitative differences were observed in hypothalamic testosterone metabolism or in the pituitary 17 beta-hydroxysteroid dehydrogenase activity. Testosterone administration to intact male rats decreased the pituitary 5 alpha-reductase activity and LH, while administered to castrated rats, it was able to suppress totally the castration-induced increase of the 5 alpha-reductase activity and of the gonadotropin secretion. The drastic decrease of the plasma levels of testosterone, observed after a prolonged treatment with GnRH, goserelin or Org 30276 was not accompanied by an increased pituitary 5 alpha-reductase activity. Injected to castrated rats, it was observed that the castration-induced increase of the pituitary 5 alpha-reductase was further stimulated by GnRH, totally suppressed by goserelin and partially suppressed by Org 30276. Concomitant administration of goserelin or Org 30276 and testosterone propionate to castrated rats resulted in a further decrease of the pituitary 5 alpha-reductase activity, compared to the castrated, GnRH-analogue treated rats. These data indicate that the pituitary 5 alpha-reductase enzyme system is controlled by both direct steroidal and indirect GnRH-mediated mechanisms.

Metabolism of [4-14c]testosterone and precursors by homogenates of rat testes after chronic LHRH-treatment

Adult male rats were injected daily for 8 days with an LHRH agonist. Twenty-four hours after the last injection testes-homogenates were incubated in the presence of a 4-14C-labeled steroid, either progesterone, 17 alpha-hydroxyprogesterone, dehydroepiandrosterone, androstenedione or testosterone. The activity of several enzymes involved in the androgen biosynthetic pathway was inferred from the amount of metabolites produced under these conditions. After LHRH-treatment a significant increase in the 17,20-lyase activity was observed without any significant change in the activity of 17 alpha-hydroxylase, 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase and 17 beta-hydroxysteroid dehydrogenase. The results of the experiments indicate that the decreased testosterone secretion observed in rats after chronic LHRH-administration is not due to an inhibition of the enzyme-systems studied.

Testosterone metabolism by incubated rat testes after chronic LHRH treatment

Adult male rats were injected 4 or 8 days with LHRH agonist. After sacrifice the testes were incubated in vitro with or without [4-14C]testosterone. After LHRH-administration the endogenously produced amounts of testosterone and of 7 alpha-hydroxytestosterone, the main testosterone metabolite normally found on incubation of adult rat testes, were drastically reduced when compared with controls. hCG, injected to rats 2 h before sacrifice, increased steroid production. In the LHRH-treated rats, however, the amounts of testosterone and of 7 alpha-hydroxytestosterone produced were much less while an important formation of 5 alpha-androstanediol was observed. The testes of LHRH treated rats metabolized [4-14C]testosterone to a large extent to 5 alpha-reduced and unextractable metabolites while the formation of 7 alpha-hydroxylated metabolites was much reduced. It is concluded that prolonged LHRH treatment provokes not only a depression of the testosterone production but has also an influence on the testicular metabolism pattern of testosterone resulting in a proportionally increased production of 5 alpha-reduced steroids and unextractable metabolites while the formation of 7 alpha-hydroxylated steroids is inhibited.

Influence of a prolonged tamoxifen administration on steroidogenesis by incubated rat testes

Tamoxifen was administered i.m. for 9 days to adult male rats in a daily dose of 100 micrograms or 1 mg. The treatment resulted in a significant reduction of the plasma levels of testosterone and LH, without modification of the plasma levels of FSH and of the testes weight. Upon incubation, the testes from the tamoxifen-treated rats produced less testosterone and 7 alpha-hydroxytestosterone, but metabolized [4-14C]testosterone in the same way as the control animals. Small doses of hCG (0.5 i.u. for 9 days) were unable to modify the tamoxifen effect on the testicular function, while tamoxifen significantly inhibited the increase of the plasma levels of testosterone induced by the administration of moderate doses of hCG (1.5 i.u. or 2.5 i.u. for 9 days) to hypophysectomized rats. Tamoxifen treatment, however, did not modify significantly the reactivity of the testes towards high doses of hCG (10 i.u.), administered either 2 h before sacrifice or for 9 days. It is concluded that a prolonged administration of tamoxifen in the rat has, besides an indirect effect resulting from a decrease of the LH levels, a direct inhibitory influence on the testicular testosterone formation, which can be reversed by high doses of hCG.