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Little is known concerning the sensitivity of the reproductive tract to ethanol as a function of development. The present study was conducted to evaluate the action of chronic ethanol ingestion on sexual maturation of the male. Mice were given free access to liquid diets containing 5% (v/v) ethanol for either 29 or 43 days, starting at age 20 days. Controls were given liquid diets in which isocaloric sucrose replaced the ethanol. Daily diet consumption and peak blood ethanol levels were highest during the first 2 weeks of treatment, dropping thereafter to adult levels of approximately 680 ml/kg body weight and 160 mg/dl respectively. Plasma testosterone levels were depressed by ethanol throughout treatment, the reduction being somewhat greater when measured during week 6 of treatment (average = 74% inhibition) as compared to either week 2 (36%) or week 4 (25%). Average weights of testes, epididymides and seminal vesicles were depressed by 24% (P less than 0.002), 16% (P less than 0.005) and 13% (NS), respectively, after 29 days. Testicular development was also impaired in ethanol-treated animals after 29 days. Tunica albuginea thickness and seminiferous tubule diameter were decreased (by 31%, P less than 0.05; and 16%, P less than 0.01 respectively), whereas desquamation of immature germ cells and inactive tubules were increased (325 and 780% respectively; P less than 0.01). Quality of spermatogenesis was poorer in ethanol-treated animals (P less than 0.05). Also observed were decreased sperm motility (62% inhibition, P less than 0.01) and capacity to fertilize (decreased by 67%, P less than 0.01), and an increase in the incidence of morphologically abnormal spermatozoa (by 163%, P less than 0.001). Semen volume was lower (reduced by 57%, P = 0.05), as was the total number of motile ejaculated spermatozoa (reduced by 81%, P less than 0.05). After 43 days treatment, improvement was noted in all indices of fertility except for the number of motile ejaculated spermatozoa. Significant differences persisted only for dysmorphic spermatozoa and volume and sperm count of electroejaculated semen. These data suggest that ethanol ingestion during pubertal development can delay several aspects of sexual maturation in the male.

In vitro studies have shown that acetaldehyde is a more potent inhibitor of testicular steroidogenesis than ethanol. The present study examined the in vivo role of acetaldehyde in ethanol-induced reduction of testosterone by (1) determining the levels of acetaldehyde to which the testes were exposed subsequent to acute ethanol administration to mice; and (2) examining the effect of ethanol on testosterone in animals subsequent to drug pretreatment which decreased or increased ethanol-derived acetaldehyde. Ethanol-induced (3 g/kg) depression of testosterone was dependent upon gonadotropin stimulation. The increase in hCG-induced testosterone was suppressed (P less than 0.01) in ethanol- as compared to saline-treated animals [39.8 +/- 2.6 (S.E.M.) vs 28.1 +/- 2.3 ng/ml]. Pargyline (100 mg/kg) or cyanamide (8.4 mg/kg) increased (P less than 0.05) plasma and testicular acetaldehyde, while having no effect on the testosterone response to ethanol. Similarly, 4-methylpyrazole (25 mg/kg) reduced blood and testicular acetaldehyde to nondetectable levels, while having no effect on testosterone. Testicular acetaldehyde was lower (P less than 0.001) than plasma levels (14 +/- 2 vs 2.0 +/- 0.2 microM). This functional blood-testis barrier to acetaldehyde could be explained by testicular aldehyde dehydrogenases in the mitochondria (Km for acetaldehyde = 1.5 microM) and in the cytosol (Km = 123 microM) whose maximal activities totaled to more than 25-fold greater than that of testicular alcohol dehydrogenase (ADH). ADH was concentrated in the Leydig cells, while aldehyde dehydrogenase was evenly distributed in the testis. Ethanol prevented further hCG-induced rises in testosterone rather than inhibiting testosterone production to below pre-ethanol values. The above data argue against a significant role of acetaldehyde in the in vivo response of testosterone to ethanol. Ethanol appears to impair gonadotropin-testicular receptor interaction in vivo.

Temperature and ion sensitivity of human acrosin (EC 3.4.21.10) was compared to that of human trypsin. With the exception of zinc, no ion tested had significant effects on either enzyme. Zinc behaved as a noncompetitive inhibitor of both enzymes, with inhibition constants of 1.8 and 1.7 mM for acrosin and trypsin respectively. Trypsin was inhibited by the chelators EDTA and EGTA, a specific effect reversed by either calcium or magnesium. EDTA inhibited acrosin in a nonspecific manner, while EGTA was without effect. Unlike acrosin from the other species, human acrosin was unaffected by calcium or the polyamines, spermine and spermidine. Acrosin was sensitive to inhibition by preincubation temperatures above 5 degrees C; trypsin, however, was stable to preincubation temperatures up to 60 degrees C. Hydrolysis of N-alpha-benzoyl-L-arginine ethyl ester was more efficiently catalyzed by trypsin (6800 cal/mol) than by acrosin (9511 cal/mol).

Proacrosin was purified from acid extracts of human spermatozoa by concanavalin A precipitation and Bio-Gel P-100 chromatography. Two molecular weight forms of proacrosin were obtained, a major one with a Mr of 70,000-71,000 and a minor one with a Mr of 47,000-53,000. In contrast to sperm extracts, the purified forms of proacrosin were free of acrosin inhibitor(s) and nonzymogen acrosin. By modulating pH, ionic strength and temperature, the activation of proacrosin in sperm extracts was compared to only the major form of purified proacrosin, since it seemed to be the source of the lower molecular weight form of proacrosin. In both preparations, proacrosin activation occurred maximally over a broad pH range (7.6-8.8 for purified proacrosin and 7.6-9.6 for extract). Additionally, an ionic strength of 0.1 and above caused a decrease in proacrosin activation in both preparations. Similarly, proacrosin was sensitive to short incubation periods at 45 degrees C and above which caused a decrease in the amount of proacrosin found in both preparations.