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1.
Biol Reprod ; 102(1): 92-101, 2020 02 12.
Article in English | MEDLINE | ID: mdl-31504198

ABSTRACT

A crucial function of the epididymis is providing a surface glycocalyx that is important for sperm maturation and capacitation. Defensins are antimicrobial peptides expressed in the epididymis. In the macaque epididymis, defensin beta 126 (DEFB126) is important for sperm motility, however, it is not known whether this is the case in humans. The objectives were to determine: (1) if DEFB126 on human ejaculated sperm was correlated with sperm motility in fertile and infertile men, (2) that recombinant DEFB126 could induce immature sperm motility in vitro. Immunofluorescence staining indicated that the proportion of DEFB126-positive sperm was significantly higher in motile sperm. Furthermore, the proportion of DEFB126-labeled sperm was positively correlated with sperm motility and normal morphology. Additional studies indicated that the proportion of DEFB126-positive spermatozoa in fertile volunteers was significantly higher than in volunteers with varicocele, and in infertile volunteers with semen deficiencies. To determine the role of DEFB126 on sperm motility, the DEFB126 gene was cloned and used to generate recombinant DEFB126 in H9C2 cells (rat embryonic heart myoblast cells). Deletion mutations were created into two regions of the protein, which have been linked to male infertility. Immotile testicular spermatozoa were incubated with cells expressing the different forms of DEFB126. Full-length DEFB126 significantly increased motility of co-cultured spermatozoa. However, no increase in sperm motility was observed with the mutated forms of DEFB126. In conclusion, these results support the notion that DEFB126 is important in human sperm maturation and the potential use of DEFB126 for in vitro sperm maturation.


Subject(s)
Fertility/physiology , Infertility, Male/metabolism , Sperm Motility/physiology , beta-Defensins/metabolism , Adult , Epididymis/metabolism , Humans , Infertility, Male/genetics , Male , Middle Aged , Sperm Maturation/physiology , Spermatozoa/metabolism , Young Adult , beta-Defensins/genetics
2.
Immunol Invest ; 43(7): 617-26, 2014.
Article in English | MEDLINE | ID: mdl-24927491

ABSTRACT

Seminal plasma and follicular fluid (FF) cytokine analysis are valuable tools for diagnoses and validation of therapeutic approaches for improving the chance of conception. Despite the initial discovery over a decade ago, the IL-17 family has not received much attention in the case of infertility. In this study, we analyzed the level of IL-17A in seminal plasma, follicular fluid and blood serum of infertile patients with different clinical diagnoses by Enzyme Linked Immunosorbent Assay (ELISA). The results showed that the level of IL-17A was higher in seminal plasma and blood serum of varicocele patients than the control group. The level of this cytokine was higher in follicular fluid of endometriosis, polycystic ovary syndrome (PCOS) and tubal factor patients than the control group. A similar elevation in IL-17A level was observed in blood serum of these patients. Furthermore, there was a correlation between the numbers of meiosis I (MI) oocytes and the level of blood serum and follicular fluid IL-17A in PCOS patients. Our data suggest a putative role of IL-17A in mediating these conditions and may have possible applications in the development of more effective diagnostic tools and therapeutic treatments for human reproductive disorders.


Subject(s)
Follicular Fluid/metabolism , Infertility, Female/metabolism , Infertility, Male/metabolism , Interleukin-17/metabolism , Semen/metabolism , Adult , Endometriosis/metabolism , Female , Humans , Interleukin-17/blood , Male , Polycystic Ovary Syndrome/metabolism , Varicocele/metabolism , Young Adult
3.
Iran J Reprod Med ; 10(4): 343-8, 2012 Jul.
Article in English | MEDLINE | ID: mdl-25246896

ABSTRACT

BACKGROUND: In vitro maturation (IVM) of human oocytes is an emerging procedure quickly incorporated into the world of assisted reproductive technologies. As an effective method of in vitro maturation, several studies have reported the critical role of differentions on activating the complex process involved in both gamete maturation and fertilization. OBJECTIVE: In this study, we supplemented a chemically defined medium with different combinations of selenium, calcium and calcium ionophore concentrations to obtain the best rate of human oocytes maturation, survival, and fertilization. MATERIALS AND METHODS: As an experimental study, Three combinations of [selenium (5 µg/ml), calcium (5 µg/ml) and calcium ionophore (1 µg/ml)], [selenium (10 µg/ml), calcium (7 µg/ml) and calcium ionophore (2 µg/ml)] and [selenium (15 µg/ml), calcium (10 µg/ml) and calcium ionophore (5 µg/ml)] added to the chemically defined medium and the morphology of oocytes assessed after 22-24 hours in vitro maturation of the oocytes. RESULTS: The highest percentage of MII (meiosis II) oocytes (68%), developing beyond the morula (20.1%) and the blastocyst formation (11.1%) observed in oocytes treated with 15µg/ml selenium, 10µg/ml calcium and 5µg/ml calcium ionophore. Moreover, we showed the significant rate of survival in each three combinations after 36, 72 and 96 hours. CONCLUSION: Maturation and activation of oocytes may be triggered by changes in intracellular ion concentrations as second messengers in signal transduction pathways. Here, we received the highest percentage of in vitro maturation and fertilization among three combinations of selenium, calcium and calcium ionophore treatments. Using this combination of ions beside other factors might be useful for the enrichment of the human oocytes IVM medium.

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