RESUMO
Phospholipase C zeta (PLCζ) is an important inducer of Ca2+ oscillations in mammalian sperm. To explore the influence of PLCζ on early embryonic Ca2+ fluctuations during sperm-egg binding, this study used PLCζ from sheep sperm to construct an early embryonic Ca2+ fluctuation model. First, sheep MII oocytes were cultivated and screened using microinjection technology. Then, a pEGFP-N1-PLCζ plasmid was constructed to activate oocytes in the test group. Ionomycin combined with 6-Dimethylaminopurine (6-DMAP) was used for the control group to explore the effects on early embryonic development and regulation of Ca2+ fluctuations during development. The results demonstrated that both the PLCζ and ionomycin combined with 6-DMAP activation methods induced sheep oocyte parthenogenetic activation and development in early embryos. In comparisons, the cleavage rate of ionomycin combined with 6-DMAP activation was significantly higher than that of PLCζ (60.9% ± 19.4% vs 76.1% ± 0.7%, respectively; p < 0.001), and the blastocyst rates were 16.2% ± 0.62% and 21.1% ± 0.92%, respectively (p < 0.05). Additionally, when comparing the distribution of Ca2+ in early embryos at different stages, Ca2+ in both treatment groups was mainly distributed in the cytoplasm, but the temporal pattern of Ca2+ fluctuations differed. PLCζ resulted in Ca2+ peaks that appeared at the cleavage and morula stages of early embryos, and Ca2+ returned to normal levels at the morula stage. However, the Ca2+ concentration after ionomycin combined with 6-DMAP activation was always much higher than that with PLCζ, and its single peak appeared later than in the PLCζ group. In summary, the PLCζ gene promoted stable regulatory effects on Ca2+ fluctuations at different stages during early embryonic development.
Assuntos
Sêmen , Fosfolipases Tipo C , Animais , Embrião de Mamíferos/fisiologia , Feminino , Ionomicina/farmacologia , Masculino , Mamíferos , Oócitos/fisiologia , Gravidez , Ovinos , Espermatozoides , Fosfolipases Tipo C/metabolismoRESUMO
Infertility is a serious public health problem worldwide. Molybdenum (Mo) plays an important role in maintaining normal metabolism. To explore the therapeutic efficacy of molybdenum (Mo) on male infertility, 90 mice were randomly divided into control, busulfan and busulfan + Mo groups. The male mice in the busulfan and busulfan + Mo groups were exposed to busulfan (20 mg/kg body weight) with a single intraperitoneal injection to establish the infertility model. The sterile mice were successfully obtained 30 days after busulfan exposure. Then, the male mice in the busulfan + Mo group were given drinking water containing 20 mg/L Mo continuously for 42 days. At 72 Day after treatment, 30 mice in the three groups were tested for various indices, and 60 mice were mated with females in spontaneous estrus. Mo significantly reversed the thinner seminiferous tubules and disappeared tubule and germ cells. Mo also normalized previously abnormal levels of testosterone, estradiol, luteinizing hormone, superoxide dismutase, lactate dehydrogenase, malondialdehyde. Furthermore, expression levels expression of Bax, Bcl-2, caspase-3 and caspase-9 returned to control levels; and finally, Mo-treated sterile mice obtained offspring with normal number and gender ratio. These results suggested that Mo at 20 mg/L had a significant therapeutic effect on reproductive dysfunction in sterile mice. Its mechanism could via repair of damaged testicular structures, regulation of abnormal reproductive hormone levels, decreased oxidative stress or and resistance to cell apoptosis. Mo may be a new candidate medicine for treatment of male infertility.