Journal of Andrology
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Published-Ahead-of-Print March 21, 2007, DOI:10.2164/jandrol.106.001958

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Impact of Ca2+ Flux Inhibitors on Acrosome Reaction of Hamster Spermatozoa

Huai L. Feng *, Yi. B. Han , Avner. Hershlag , and Lei. J. Zheng

* To whom correspondence should be addressed. E-mail: hfeng{at}nshs.edu.

Ca2+ plays a predominant role in the regulation of critical functions of spermatozoa, such as capacitation, acrosome reaction (AR) and fertilization. While there is consensus that Ca2+ is essential, researchers have reported conflicting results as to what happens to Ca2+ flux across the sperm membrane during capacitation and AR. The purpose of the present study is to further delineate the function of Ca2+ channels and their role in sperm capacitation and AR. Epididymides were obtained from healthy adult male hamsters. Spermatozoa were washed with modified Tyrode's medium supplemented with 0.3% BSA, adjusted to 4.5 x 107 motile sperm/ml, and incubated in 200µl droplets for 4 hrs at 37°C in the presence of either the trifluoperazine (TFP, calmodulin inhibitor at 25, 50, 100, 150 nM, respectively), or verapamil (VP, Ca2+-channel inhibitor at 25, 50, 100, 150 nM, respectively), or nifedipine (NF, the voltage-dependent Ca2+-channel inhibitor at 50, 100, 200, 400nM, respectively). Spermatozoa were assessed for AR by using Coomassie Brilliant Blue staining techniques. Results indicated that incubation of sperm with Ca2+ channel inhibitors for 4 hours significantly reduced the AR in the study groups (TFP: 88 ±2.3%, 65 ±2.0%, 60 ±2.2%, 54 ± 2.2%, respectively; VP: 45 ±1.3%, 23 ± 1.2%, 12 ±1.0%, 8 ±0.6%, respectively; NF: 11 ± 0.8%, 9 ±0.3%, 7.0 ±0.1%, 6.0 ±0.1%, respectively) compared to control group (95 ± 3.0% AR, P<0.05). However, increasing the concentration of TFP and NF did not result in further suppression of the AR. In summary, Ca2+ channel antagonists suppress the AR. Key Words: Calcium, spermatozoa, acrosome reaction, inhibitor


Key words: Fertilization • Infertility • Sperm




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