Effect of alcoholic extract of Lepidium meyenii （Maca） on testicular function in male rats

Aim: To evaluate the effect of the alcoholic extract of Lepidium meyenii (Maca) on the spermatogenesisin male rats. Methods: In Holtzman rats, Maca alcoholic extract (5 %) was given by oral route at doses of 48 mg/day or 96 mg/day for 7 days, 14 days and 21 days. Testicular function was assessed by measurements of lengths of different stages of seminiferous epithelia and by epididymal sperm count. Results: Ethanolic extract of Maca increased the length of stages IX-XI of seminiferous epithelium at treatment day 7, day 14 and day 21. Progression of spermatogenesis was evident only after day 21 when lengths of stages XII-XIV of seminiferous epithelium were increased; at day 7 and day 14, no important change in spermatogenesis was observed. Epididymal sperm count was increased with 48 mg/day at all times. With 96 mg/day an increase in sperm count was observed at day 7, but it was reduced at day 14 and day 21 of treatment. Serum testosterone levels were not affected. Conclusion: The alcoholic extract of Maca activates onset ant progression of spermatogenesis at 48 mg/day or 96 mg/day in rats.     

Possible role of elongated spermatids in control of stage-dependent changes in the diameter of the lumen of the rat seminiferous tubule

Adult male rats were treated with a single dose of 650 mg/kg methoxy acetic acid to deplete the seminiferous tubules specifically of pachytene and later spermatocytes. The effect of this treatment and the subsequent maturation-depletion of later germ cell types on the diameter of the seminiferous tubule and its lumen and the area of the seminiferous epithelium were studied in relation to the stages of the spermatogenic cycle. At 21 days after methoxy acetic acid treatment, the diameter of the tubule and the area of the epithelium were reduced below control values at all stages, consistent with the reduced number of early (stage VIII) or late (all other stages) spermatids. Unexpectedly, diameter of the lumen was also reduced at all stages other than VIII, and especially at stage VII. In controls, lumen diameter at stages VII and VIII was increased by approximately 50% compared with earlier and later stages. In rats treated 21 days previously with methoxy acetic acid no change occurred at stage VII (lacking elongated spermatids) while a normal increase did occur at stage VIII (lacking round but not elongated spermatids). At earlier times after methoxy acetic acid treatment when stage VII tubules were depleted of pachytene spermatocytes alone (3 days) or together with early spermatids (7 days), the diameter of the lumen was not significantly different from the control value. These data suggest that lumen diameter may be regulated by elongated spermatids, especially at stages VII and VIII.     

Effect of high altitude exposure on spermatogenesis and epididymal sperm count in male rats

The present study was designed to determine the effect of exposure to high altitude on spermatogenesis using transillumination technique and sperm count in male rats. In addition, the effect of oral intubation for intragastric administration of vehicle on testicular parameters in adult male rats in a schedule of 42 days was assessed. Male rats were exposed to Cerro de Pasco (Peru) at 4340 m for 3, 7, 14, 21, 28, 35 and 42 days resulting in a modification of the pattern of the seminiferous tubule stages. At day 3, stages I, IV-V, VI, VII and IX-XI were relatively shorter at high altitude than at sea level. At day 7, stages VIII, IX-XI, XII and XIII-XIV were reduced. At day 14, stages VII, VIII and IX-XI were reduced. At day 21 and 28, stages VIII, XII and XIII-XIV were significantly increased at high altitude. At day 35 an increase in stage XIII-XIV was observed. At day 42, stages II-III, IX-XI and XII were significantly increased at high altitude. Epididymal sperm count was significantly reduced at day 7 of exposure to high altitude and maintained low levels with respect to sea level up to 42 days. In conclusion, high altitude exposure affects spermatogenesis, particularly onset of mitosis and spermiation. This in turn affects epididymal sperm count.     

The effect of selective destruction and regeneration of rat Leydig cells on the intratesticular distribution of testosterone and morphology of the seminiferous epithelium

This study was designed to explore the relationship between the intratesticular distribution of testosterone and spermatogenesis by completely destroying the Leydig cells of mature male rats with injection of a single i.p. dose of ethane dimethanesulphonate. After such treatment, testosterone levels in serum, testicular interstitial fluid, seminiferous tubules, and whole testis declined significantly 6 to 24 hours after injection and fell below assay detection limits between 3 and 7 days. At 3 and 7 days, serum LH and FSH levels rose significantly and remained elevated up to 4 and 6 weeks, respectively, in comparison with vehicle-treated controls. Leydig cells disappeared from the interstitium by day 3, but between 2 and 4 weeks postinjection a new generation of fetal-like Leydig cells repopulated the testicular interstitium and, during weeks 6 to 10, were transformed into, or replaced by, Leydig cells with an adult type of morphology. Histologic examination of the seminiferous tubules showed progressive disruption of spermatogenesis between 3 and 14 days post-ethane dimethanesulphonate. The first histologic sign of spermatogenic damage was noted at day 3, with the occurrence of stage-specific degenerating pachytene primary spermatocytes at stages VII to VIII of the spermatogenic cycle. On day 7, these cells and degenerating round, or step 19, spermatids often were observed during stages VII to XI, although qualitatively normal spermatogenesis also was seen in these and all other stages of the cycle. Maximum impairment of spermatogenesis occurred 2 weeks post-ethane dimethane sulphonate, at which time the tubules commonly lacked one or more germ cell generations or, alternatively, showed accumulation of lipid inclusions, extracellular spaces, and variable numbers of degenerating germ cells. Following repopulation of the testis by Leydig cells during weeks 3 and 4, spermatogenesis recovered. By 10 weeks after treatment, qualitatively normal spermatogenesis was seen in the great majority of seminiferous tubules, although a few tubules still remained in which the germ cell complement was severely reduced, and contained only Sertoli cells and spermatogonia.(ABSTRACT TRUNCATED AT 400 WORDS)     

Marmoset spermatogenesis: organizational similarities to the human

The common marmoset (Callithrix jacchus), a small New World primate of high fecundity, is widely used in reproductive research. The aim of the present study was to determine the organization of the germ cells within the seminiferous epithelium, the duration of the spermatogenic cycle and the number of spermatogonial mitoses. Antibodies to cAMP response element modulator (CREM) and proliferating nuclear cell antigen (PCNA) and a cRNA directed against protamine P2 and morphological criteria were used to discriminate between stages of the spermatogenic cycle. Plastic sections were used to document the cell associations present in each of the nine stages of spermatogenesis. Up to five such stages could be observed within individual cross-sections of seminiferous tubules. Based on the pattern of incorporation of bromodeoxyuridine the length of the spermatogenic cycle was estimated to be 10 days and the duration of spermatogenesis to be 37 days. Four mitotic divisions were noted in spermatogonia. It is concluded that the organization of spermatogenesis in the marmoset has similarities to the human ('helical') and this makes the marmoset a suitable model for studies relevant to human testicular function.     

Spermatogenesis of Rat: Effect of Central Norepinephrine Synthesis Inhibition by Diethyldithiocarbamate

Spermatogenesis and steroidogenesis of male Wistar rats under the influence of a decreased central NE level was studied. Inhibition of NE synthesis was produced by chronic injection (7 days) of DDC, a Dopamine hydroxylase blocker, which decreased brain NE, increased brain DA without significantly affecting brain 5-HT. Rats were sacrificed on the day after cessation of treatment (8th day) and after an interval of 13 days following the cessation of treatment (21st day). The time interval of 13 days is equivalent to one cycle of the seminiferous epithelium in Wistar strain rats. A degenerative change (reduced spermatid count) in the spermatogenic pattern at stage-VII of the cycle of the seminiferous epithelium was observed in the rats sacrificed on the 8th day, the degeneration being much greater in the rats sacrificed on the 21st day. Rats sacrificed on the day after cessation of DDC treatment revealed an inhibition of plasma testosterone level. Such change was not observed in the rats sacrificed after the interval of 13 days following DDC treatment. Human chorionic gonadotropin supplementation in the rats sacrificed on the 21st day partially restored the spermatogenesis towards the vehicle treated control. The inhibition of spermatogenesis and steroidogenesis reflects a decrease in the pituitary gonadotropin secretion under the influence of a decreased NE synthesis in brain, following chronic treatment of DDC.     

The selective removal of pachytene spermatocytes using methoxy acetic acid as an approach to the study in vivo of paracrine interactions in the testis

Testicular weight and morphology, serum gonadotropins, intratesticular levels of testosterone and ABP levels in testicular interstitial fluid were studied in adult rats at intervals of 1 to 70 days after a single oral dose of 650 mg/kg methoxy acetic acid. At 3 days, this treatment resulted in the selective loss or depletion of pachytene and later spermatocytes from seminiferous tubules at all stages other than VIII to XI of the spermatogenic cycle. At later times this lesion was expressed as an absence mainly of round (14 days) or elongated (21 days) spermatids from the majority of seminiferous tubules. Other than these changes, spermatogenesis did not appear to be affected by treatment and was qualitatively normal in all tubules at 70 days after treatment. As deduced from cell counts at 3 days posttreatment, the initial action of methoxy acetic acid was restricted to late zygotene spermatocytes (stage XII) and pachytene spermatocytes at all stages other than early- to mid-stage VII. Levels of FSH in serum and those of ABP in testicular interstitial fluid indicated that Sertoli cell function was altered in rats treated with methoxy acetic acid. Both were increased at 1 to 3 days posttreatment, returned to normal at 7 to 14 days but were increased again at 21 days before finally returning to control levels at 28 days. In contrast, the levels of testosterone in serum, isolated seminiferous tubules and testicular interstitial fluid were unaffected by treatment, as also were the serum levels of LH. The two periods of increase in FSH and ABP levels coincided with the times of greatest decrease (approximately 20%) in testicular weight, and may be related either to the type of germ cell missing from the affected tubules and/or to the stage of the cycle of the affected (or unaffected) tubules. These data suggest that chemicals such as methoxy acetic acid may prove useful in the study of paracrine interactions in vivo.     

Cycloheximide prevents production of arresting, a fraction of 30-50 kDa obtained from seminiferous tubule conditioned medium

Aim: To evaluate the effect of a protein synthesis inhibitor cycloheximide on arresting activity in spermatogenesis and sperm count in male rats. Methods: The study used seminiferous tubule (ST) segments from adult rats cultured in vitro with or without cycloheximide to condition culture media, which have been concentrated, size fractioned (30-50 kDa) and administered 7 days to adult rats by intraperitoneal injections. The effects on testicular and epididymal weights, spermatogenesis and epididymal sperm count were determined. Results: The fraction (30-50 kDa), named arresting, obtained from the culture without cycloheximide decreased testicular and epididymal weights (P＜0.01) and reduced the epididymal sperm count significantly. Study of the spermatogenic cycle by transillumination showed spermatogenic arrest at stage VⅡ in rats treated with arresting compared to that observed in controls. The length of stage VⅡ in the group receiving the seminiferous tubules culture media with cycloheximide (30-50 KDa CHX-STCM fraction) was similar to control. Conclusion: The difference in the effect may be the result of the presence or absence of arresting, a protein secreted by the tubules. (Asian J Androl 2004 Dec;6:359-364)     

Evaluation of the effect of selective germ cell depletion on subsequent spermatogenesis and fertility in the rat

Rats were treated with a single high dose of methoxy acetic acid (MAA; 650 mg/kg) specifically to deplete seminiferous tubules of pachytene and later spermatocytes. The impact of this selective depletion on subsequent spermatogenesis, sperm output and fertility was then evaluated at intervals ranging from 3 days to 10 weeks. Cauda epididymal sperm number was reduced progressively beyond 2 weeks post-treatment and reached a nadir at 5-6 weeks (28-34% of control values) before recovering progressively back to control levels at 10 weeks. Sperm motility was reduced significantly at 4-7 weeks post-treatment with a nadir at 6 weeks (35% of control values). Thus, at 5-6 weeks after MAA treatment, motile sperm output was reduced by 82-88%. Despite these changes, there was little evidence for infertility in the majority of treated males during a serial mating trial. Evaluation of seminiferous tubule morphology combined with germ cell counts at stage VII of the spermatogenic cycle confirmed that, initially, MAA induced the specific loss of pachytene and later spermatocytes at all stages other than early to mid stage VII. Maturation depletion of germ cells at later intervals was consistent with the initial effects of MAA, although at 21 days post-treatment a number of unpredicted (? secondary) changes in spermatogenesis were observed. These were (a) a reduction in number of pachytene spermatocytes at late stage VII/early stage VIII, (b) retention of sperm at stages IX-XIV, and (c) increased degeneration of pachytene spermatocytes and round spermatids at stage VII and of secondary spermatocytes at stages XIV-I. Whilst none of these changes was severe, together they probably accounted for the unexpectedly prolonged drop in sperm output. It is concluded that whilst deleterious changes in spermatogenesis may occur secondarily following MAA treatment, for the most part spermatogenesis proceeds normally and fertility is largely maintained despite a massive but transient decrease in sperm output.     

Repeated interruptions of the testicular blood flow do not have long-term effects on spermatogenesis in the ram

Summary. The effect of repeated interruptions of the testicular blood flow on spermatogenesis was studied in mature Texel rams. Reversible interruption of the blood flow was achieved by an inflatable occluder, placed around the testicular artery at the level of the spermatic cord. In eight testes the blood flow was successfully interrupted six times for 1 h within 3 weeks and in 14 testes nine times for 1 h within 3 weeks. Nine weeks after the last blood flow interruption spermatogenesis was evaluated in histological sections of the testes. Both after six and nine blood flow interruptions a qualitatively complete epithelium was found in at least 90% of the seminiferous tubules. Cell counts in stages VII and VIII of the spermatogenic cycle revealed a slight decrease of spermatocytes and spermatids in the tubules with a complete epithelium after nine occlusions, which was only statistically significant for Preleptotene Spermatocytes. After six occlusions the numbers of all cell types were at or even slightly above control levels. These results show that repeated periods of ischaemia for 1 h do not result in conspicious long-term damage to spermatogenesis.