چکیده (انگلیسی):

Objective: To determine the effect of different concentrations of heparin, caffeine or
calcium ionophore and incubation time on motility, hyperactivity (HA) and acrosome
reaction (AR) of ram sperm in vitro.
Methods: Semen samples were collected by artificial vagina from three mature ram.
Split fractions (0.1 mL) of the pooled semen were layered under 1 mL of S-TALP medium
supplemented with either heparin (10, 25, 50, 75 and 100 mg/mL), caffeine (0.5,
0.75, 1, 2 mg/mL), or calcium ionophore A23187 (0.5, 0.75, 1, 1.55 mM/mL). Individual
motility, hyperactivity percentage and acrosomal status were recorded at 0, 1, 2, 3 and 4 h
post-incubation for all treatments and control. Moreover, they were examined for ability
to fertilize sheep oocyte in vitro.
Results: Heparin, caffeine and calcium ionophore A23187 at a concentration of 75 mg/
mL, 1 mg/mL, and 1.55 mM/mL respectively can be used as a protocol to provide the
best results for in vitro cap citation and acrosome reaction in ram. The penetration rates of
rate of oocytes inseminated with spermatozoa treated with calcium ionophore A23187,
and heparin were higher as compared with caffeine. Moreover, heparin achieved higher
fertilization rates but without significant difference with others.
Conclusion: The best concentration of heparin, caffeine and ionophore A23187 are
75 mg/mL, 1 mg/mL, 1.55 mM/mL for 3, 1, 4 h incubation respectively and can be used
for in vitro fertilization in sheep.