Changes in XXXX immunocytochemical localization associated with capacitation and acrosomal exocytosis of ram spermatozoa.

@entity137 @entity563 @entity2667 @entity3282 @entity13609 @entity5993 @entity3576 @entity7914 @entity460 @entity3365 @entity141 @entity669 @entity1889 @entity6106
The aim of this study was to determine the localization of @entity13609 ( @entity13609 ) in ram sperm and the possible changes during in vitro capacitation (CA) and the ionophore-induced @entity137 ( @entity137 ). Likewise, changes in intracellular @entity460 levels ([ @entity141 ](i)) were also analysed by using flow cytometry. CA was induced in vitro in a medium containing BSA, @entity3282 , @entity5993 , and @entity137 by the addition of the @entity460 ionophore @entity3365 . The acrosomal status was assessed by the @entity7914 -fluorescence (CTC) assay. Flow cytometry (FC) analyses were performed by loading samples with @entity3576 AM, that emits fluorescence at a high [ @entity141 ](i), combined with @entity2667 (PI) that allowed us to discriminate sperm with/without an integral plasma membrane both with high/low [ @entity141 ](i). Immunocytochemistry localized @entity13609 to the flagellum, and some sperm also contained @entity13609 in the head (equatorial and post-acrosomal regions). CA and @entity137 resulted in a slight increase in the post-acrosomal labelling. The treatment of sperm with increasing concentrations of two @entity13609 antagonists, W7 and @entity6106 (CZ), accounted for an increase in capacitated and acrosome-reacted CTC-sperm patterns. CZ induced a significant reduction in the content of three protein @entity669 -phosphorylated bands of approximately of 30, 40 and 45kDa. However, W7 showed no significant effect at any of the studied concentrations. Neither of them significantly influenced protein @entity563 and @entity1889 phosphorylation. FC analysis revealed that the main subpopulation in the control samples contained 70% of the total sperm with integral plasma membrane and a medium [ @entity141 ](i). After CA, 67.1% of the sperm preserved an integral membrane with a higher [ @entity141 ](i). After @entity137 , only 7.2% of the total sperm preserved intact membranes with a very high [ @entity141 ](i). These results imply that @entity13609 appears to be involved in ram sperm capacitation, and both treatments increased its localization in the post-acrosomal region.
