Study On Induction Of Acrosome Reaction And Regulation Of Ions Transport Of Wapiti(Cervus Elaphus) Sperm Using Progesterone & γ-Aminobutyricacid

In vitro fertilization(IVF) is the third revolution in animal reproduction field following artificial insemination(AI) and embryo transplant, which has wide development foreground and important practical signification. Sperm capacitation in vitro and acrosome reaction(AR) were premise and basic of IVF. So study on sperm capacitation and AR helps to the development of IVF. To choose a set of optimal system of wapiti(cervus elaphus) sperm AR induced by progesterone(P4) & γ-aminobutyricacid(GABA) and clarify primarily the mechanism of wapiti sperm AR, we studied induction of AR and regulation of Ca²⁺, Mg²⁺, Na⁺, K⁺, Cl^- and total F transport of wapiti sperm, using the method of twice incubation. These results showed that:1. P₄ at 0.5μmol/L¹000μmol/L could enhance the ratio of capacitation wapiti sperm AR. The effects of 1μmol/L P₄ and 10μmol/L P₄ were significant(P<0.05). The maximal stimulatory effect was observed with 10μmol/L P₄(74.44±0.71%).2. Incubated for 10²0min by P4 at 10μmol/L, the wapiti sperm achieved higher ratio of AR(upper 72.41%), with a maximal response in incubated for 10min(78.40%).3. GABA at 0.05mmol/L².5 mmol/L could enhance the ratio of capacitation wapiti sperm AR. The maximal stimulatory effect was observed with 0.5mmol/L GABA(77.94±6.53%) (P<0.01).4. Incubated 1³0min by GABA at 0.5mmol/L, the wapiti sperm achieved higher ratio of AR(upper 70%), with a maximal response in incubated for 10min(77.52±2.96%).5. The cooperation of P4 and GABA could accelerate the AR of capacitation wapiti sperm, with the optimal effect in 1μmol/L P₄ +2.5mmol/L GABA.6. AR of capacitation wapiti sperm was accelerated by progesterone and GABA, but the AR of uncapacitation wapiti sperm was riot(P>0.05).7. Progesterone and GABA had no significant effects on wapiti sperm capacitation(P>0.05). But extending the incubating time of capacitation had significant effect on the AR induced b> GABA(P<0.01).8. The concentrations of Ca²⁺, Mg²⁺, Na⁺, K⁺, Cl^- and total P in culture medium showed changes within 15min, during AR of capacitation wapiti sperm induced by progesterone. The concentrations of Ca²⁺, Mg²⁺, Na⁺ and total P changed significantly(P<0.05) or significant(P<0.01). The concentrations of Cl^-and K⁺ fluctuated, but there were not significan(?) changes(P>0.05). Two steps of Ca²⁺ influx appeared: the first was fast and instantaneous in 60s, and the second was slow and durative after 180s.9. The concentrations of Ca²⁺, Mg²⁺, Na⁺, K⁺, Cl^- and total P showed significant (P<0.05)or significant (PO.01) changes in culture medium within 15min, during AR of capacitation wapiti sperm induced by GABA. The Ca2+ indicated efflux in the early stage of AR, but influx later. Mg2+, Na+, K+ and total P indicated influx.