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首页> 外文期刊>Andrologia >Correlation between tyrosine phosphorylation intensity of a 107 kDa protein band and A23187-induced acrosome reaction in human spermatozoa.
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Correlation between tyrosine phosphorylation intensity of a 107 kDa protein band and A23187-induced acrosome reaction in human spermatozoa.

机译：107 kDa蛋白带的酪氨酸磷酸化强度与人精子中A23187诱导的顶体反应之间的相关性。

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Summary. This study, performed using semen samples from 10 men, investigated the relationship between sperm protein tyrosine phosphorylation and acrosomal status in conditions supporting in vitro capacitation. Percoll-selected spermatozoa (cells from the 95% fraction) were incubated for 3 h at 37 degrees C under an atmosphere of 5% CO(2) in air, in a polyvinyl alcohol (1 mg ml(-1)) containing Biggers-Whitten-Whittingham's medium, nonsupplemented or supplemented with either bovine serum albumin (BSA; fatty acid free, 3 mg ml(-1)) or 2-hydroxy-propyl-beta-cyclodextrin (2-OH-p-beta-CD; 0.5, 1, 2 mmol l(-1)). Sperm suspension in each medium was split into two aliquots. The first was used to evaluate the acrosomal status by staining with the fluorescein isothiocyanate Pisum sativum agglutinin after induction of the acrosome reaction (AR) for 45 min with 10 mumol l(-1) of A23187 calcium ionophore. The second aliquot was used for sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting, followed by a densitometric analysis. Compared with the nonsupplemented medium, BSA- or 2-OH-p-beta-CD-supplementation induced an increase in both the percentage of live acrosome-reacted sperm and the tyrosine phosphorylation intensity of the main phosphorylated 107 kDa protein. A correlation between the percentage of live acrosome-reacted sperm and the 107-kDa protein phosphotyrosine intensity was observed. Therefore, the 107 kDa protein-phosphotyrosine level measurement would bring additional information to conventional semen parameters in the assessment of the human sperm functionality.
机译：概要。这项研究是使用10名男性的精液样本进行的，研究了在支持体外获能的条件下精子蛋白酪氨酸磷酸化与顶体状态之间的关系。将Percoll选择的精子（来自95％馏分的细胞）在含Biggers-的聚乙烯醇（1 mg ml（-1））中，在空气中5％CO（2）的气氛中，于37°C孵育3小时。 Whitten-Whittingham培养基，未补充或补充有牛血清白蛋白（BSA;无脂肪酸，3 mg ml（-1））或2-羟基-丙基-β-环糊精（2-OH-p-β-CD; 0.5 ，1、2 mmol l（-1））。将每种培养基中的精子悬浮液分成两等份。第一个用于通过用10摩尔I（-1）A23187钙离子载体诱导顶体反应（AR）45分钟后，通过异硫氰酸荧光素Pisum sativum凝集素染色来评估顶体状态。第二等分试样用于十二烷基硫酸钠聚丙烯酰胺凝胶电泳和免疫印迹，然后进行光密度分析。与未补充的培养基相比，BSA或2-OH-p-β-CD补充导致活的顶体反应的精子百分比和主要磷酸化的107 kDa蛋白的酪氨酸磷酸化强度增加。观察到了顶体反应的活精子百分比与107-kDa蛋白磷酸酪氨酸强度之间的相关性。因此，在评估人类精子功能时，107 kDa蛋白磷酸酪氨酸水平的测定将为常规精液参数带来更多信息。

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