Author： Liu De Yi Gordon Baker H.W.
Publisher： Oxford University Press
ISSN： 1460-2350
Source： Human Reproduction, Vol.11, Iss.3, 1996-03, pp. : 551-557
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Abstract
Acrosome reactions induced by the calcium ionophore A23187 and zona pellucid a (ZP) were studied. Sperm samples were obtained from fertile men or men with normal semen analysis and normal sperm-ZP binding. Oocytes were obtained, with the consent of the patients, after the failure of fertilization in vitro. Motile spermatozoa selected by a swim-up technique were incubated with 10 μM A23187 for 1 h, four oocytes for 2 h or solubilized ZP (4 ZP/μl) for 2 h. Spermatozoa bound to the ZP were dislodged and collected in a small volume of phosphate-buffered saline by aspirating the oocytes with a glass pipette with an inner diameter (120 μm) slightly smaller than the diameter of the oocyte. The acrosome status of the spermatozoa was determined using fluorescein-labelled Pisum sativum agglutinin. The proportion of spermatozoa undergoing the acrosome reaction on the ZP at 2 h varied over a wide range (5–99%), but the agreement between results for the same semen sample exposed to different groups of oocytes was good: the standard deviations of the differences being 9%. Pre-incubation of spermatozoa for 2 h did not increase the ZP-induced acrosome reaction. Re-incubation of ZP with the same sperm suspension for 2 h after removing ZP-bound spermatozoa from the first 2 h incubation produced a significantly lower ZP-induced acrosome reaction in the second incubation (22 ± 16%) than in the first incubation (30 ± 14%; P < 0.001,="" n="20)." there="" was="" no="" significant="" difference="" in="" the="" zp-induced="" acrosome="" reaction="" with="" oocytes="" with="" zp="" which="" had="" or="" had="" not="" been="" penetrated="" by="" spermatozoa="" during="" the="" in-vitro="" fertilization="" insemination.="" pre-incubation="" of="" spermatozoa="" with="" solubilized="" zp="" blocked="" sperm-zp="" binding.="" however,="" the="" acrosome="" reaction="" induced="" by="" solubilized="" zp="" (4="" zp/μl)="" was="" significantly="" lower="" than="" the="" acrosome="" reaction="" induced="" by="" intact="" zp="" (10="" ±="" 5="" and="" 30="" ±="" 13%="" respectively,="" n="11," p="">< 0.001),="" but="" there="" was="" a="" high="" correlation="" (spearman="" r="0.822," p="">< 0.01)="" between="" the="" results.="" on="" the="" other="" hand,="" although="" the="" average="" of="" the="" acrosome="" reaction="" was="" similar="" for="" a23187="" (42%)="" and="" for="" zp="" (43%),="" there="" was="" no="" significant="" correlation="" between="" the="" results="" for="" the="" two="" stimuli="" (n="60)." in="" conclusion,="" a="" useful="" method="" for="" assessing="" the="" zp-induced="" acrosome="" reaction="" has="" been="" developed="" using="" oocytes="" which="" failed="" to="" fertilize="" in="" vitro.="" the="" lack="" of="" a="" relationship="" between="" the="" results="" of="" the="" chemical="" (a23187)="" and="" physiological="" (zp)="" stimali="" for="" the="" acrosome="" reaction="" in="" the="" same="" subjects="" questions="" the="" biological="" basis="" of="" using="" a23187="" for="" tests="" of="" sperm="" function.="" solubilized="" human="" zp="" in="" a="" concentration="" that="" blocks="" sperm-zp="" binding="" is="" a="" less="" efficient="" inducer="" of="" the="" acrosome="" reaction="" than="" is="" intact="" zp.="" it="" is="" possible="" that="" the="" three-dimensional="" structure="" of="" the="" zp="" is="" important="" for="" induction="" of="" the="" acrosome="" reaction="" or="" that="" spermatozoa="" which="" bind="" to="" the="" zp="" are="" more="" likely="" to="" acrosome="" react="" assessment="" of="" the="" physiological="" acrosome="" reaction="" for="" diagnosis="" of="" sperm="" defects="" which="" interfere="" with="" the="" fertilization="" process="" should="" be="" concentrated="" on="" the="" spermatozoa="" which="" are="" capable="" of="" binding="" to="" the="" zp.="">
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