Actin levels fluctuate in the seminiferous epithelium at various stages during spermatogenesis in the rat

The testis is a double gland comprised of the sperm-producing seminiferous tubules and a complex endocrine interstice.The former structures generate and release whole cells(exocrine aspect of the gland)while the latter synthesize and release androgens and related hormones.The testis also has poorly understood paracrine connexions.These connexions play an important role during spermatogenesis.A key molecule within these structures is β-actin.Therefore,the present study aims at examining the expression pattern of β-actin during the various stages of the spermatic cycle in the rat.To achieve this goal,we used a combination of trans-illumination assisted microdissection of seminiferous tubules,confocal immunofluorescence and Western blot analysis.Our experiments revealed that the levels of β-actin fluctuate significantly during spermatogenesis,peaking immediately after spermiation.

High-magnification observation of seminiferous tubules through the tunica albuginea by two-photon laser scanning microscopy

Testicular sperm extraction is widely used in the treatment of male infertility in cases of non-obstructive azoospermia. Identifying spermatogenetic foci within the testes is critical for testicular sperm extraction. Two-photon laser scanning microscopy （TPLSM） is an autofluorescence-based microscopy technique that allows observation at a cellular level in the depth of fresh living tissues and does not require any histological processing （fixation or staining）. The wavelengths previously used have shown no phototoxicity on sperm. We used TPLSM to detect spermatogenetic foci in fresh mouse testicular parenchyma without disrupting the tunica albuginea. Fresh surgically retrieved testes were observed using TPLSM within 1 h after extraction. Contralateral testes for each animal were observed using standard histology. Using TPLSM we were able to observe and measure the diameter of seminiferous tubules through the tunica albuginea, similar to the histological control. Structures within epithelial tubules were also observed, although their nature has yet to be identified. TPLSM is a real-time microscopy technique that could detect spermatogenetic foci.

The peptide fraction of Bothrops jararaca snake venom induces toxicological effects on the male reproductive system after local envenomation in mice

Bothrops envenomation is complex and provokes prominent local tissue damage and systemic disturbances,but little is known about their effects on the male reproductive system.After intratesticular injection,the bioactive peptide fraction(Bj-PF)obtained from Bothrops jararaca snake venom changes the structure of different stages of the seminiferous epithelium cycle in adult mice.For the first time,we investigated whether local envenomation of Bj-PF induces toxicological effects on the male reproductive system,particularly on the seminiferous epithelium and Sertoli cells.Male adult mice were treated with 0.24 mg.kg^(-1) by intramuscular(i.m.)injection for 24 h.The testes samples were collected for morphological and morphometric evaluation.The toxicological effects of Bj-PF were also analyzed on mitochondrial metabolism and nitrite(NO2)production in 15P-1 Sertoli cell culture.Bj-PF changed the structure and function of the seminiferous epithelium,particularly the disruption of the epithelium and the presence of degenerated germ cells in the adluminal compartment,but there were no alterations in the basal compartment.Bj-PF increased the thickness of the seminiferous epithelium and decreased the lumen diameter of the tubule.Semiquantitative histological assessment of the degree of tubule degeneration revealed that Bj-PF also increased the number of hypospermatogenic tubules compared to control.Bj-PF reduced NO2 levels in 15P-1 Sertoli cells without changing the mitochondrial metabolism.Overall,the fact that Bj-PF alters the structure and function of the seminiferous epithelium suggests that bioactive peptides found in B.jararaca snake venom can have toxicological effects on the reproductive systems of affected male mice,providing new insight into the biological characteristics of snake venom and therapeutic strategies for envenomation inflammation.

Lepidium meyenii (Maca) reduces spermatogenic damage induced by a single dose of malathion in mice

Aim: To observe the effect of the aqueous extract of hypocotyls of the plant Lepidium meyenii (Maca) on spermato-genic damage induced by the organophosphate insecticide malathion in mice. Methods: Mice were treated with 80 mg kg^(-1) of malathion in the presence or absence of an aqueous extract of Maca, which was orally administered 7, 14 or 21 days after injection of the malathion. Stages of the seminiferous epithelium were assessed by transillumination on days 0, 7, 14 and 21. Results: The administration of Maca increased significantly the length of stage 鈪?on days 7, 14 and 21 of treatment compared with the controls. An increase in the length of stage 鈪?occurred on day 14 of treatment. Malathion affected spermatogenesis by reducing the lengths of stage 鈪?on day 7, stages 鈪?and 鈪?鈪?on day 14 and a recovery of stages 鈪?鈪?on day 21. The magnitude of alteration in the length of stage 鈪?produced by malathion was significantly reduced by Maca on days 7 and 14. The length of stage 鈪?was increased when Maca was administered to mice treated with malathion. Assessment of the relative length of stages of the seminiferous epithelium showed that Maca treatment resulted in rapid recovery of the effect of malathion. Conclusion: Maca enhances spermatogenesis following spermatogenic damage caused by the organophosphorous pesticide.

mpact of antioxidants on seminal vesicles function ind fertilizing potential in diabetic rats

Diabetes mellitus significantly affects the male reproduction and sexual function. In the present study, we investigated the diabetes-induced dysfunction of seminal vesicles （SVs） in the diabetes-rat model and the role of antioxidants. Streptozotocin-induced diabetes after 4 weeks caused smaller size of the organs, hypercontractility, histological abnormalities, increased concentrations of malondialdehyde in the serum and tissue, overexpression of oxidative stress markers, and cleaved caspase-3 as identified by immunohistochemistry in the SVs. In addition, diabetes resulted in deceased levels of serum testosterone and no newborns after the mating studies. Antioxidants significantly normalized all the above parameters, except for the severely decreased serum testosterone levels and the negative outcome of the mating studies. The present study gives evidence for the important role of diabetes-induced oxidative stress in the function and structure of these androgen-dependent organs. Antioxidants may be a promising supplementary therapy for diabetic male patients to alleviate ejaculatory disorders but alone is not efficient treatment for the mitigation of infertility.