Sperm acrosomal marker enzyme

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Isolation of the outer acrosomal membrane from bull sperm.

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Isolation of the outer acrosomal membrane from bull sperm.

In mammals, the success of fertilization largely depends on gamete fertility potential and consequently on what concerns sperm and oocyte quality they are both equally important. Sperm contribution to fertilization is usually estimated through evaluation of semen parameters. A loss of fertility potential associated to manipulation and preservation techniques is usually calculated based on the semen characteristics at collection and on the knowledge of the damages associated with the technique to be implemented. Assessment of sperm quality conventionally relies on microscopic evaluation of sperm parameters including total sperm count, sperm concentration, percentage of motile sperm and percentage of normal sperm morphology.
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THE IMPORTANCE OF HYDROLYTIC ENZYMES TO AN EXOCYTOTIC EVENT, THE MAMMALIAN SPERM ACROSOME REACTION

A procedure is described for subcellular fractionation of bull sperm which allows the isolation of outer acrosomal membrane without the use of detergent. After washing to remove seminal plasma contaminants, the acrosomal membrane is removed by homogenization and separated on a two-step sucrose gradient. The isolated membranes have been characterized by light and electron microscopy and enzyme analysis.
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Mammalian sperms are activated in the oviduct. This process, which involves extensive sperm surface remodelling, is required for fertilization and can be mimicked under in vitro fertilization conditions IVF. Here we demonstrate that such treatments caused stable docking and priming of the acrosome membrane to the apical sperm head surface without the emergence of exocytotic membrane fusion. The interacting membranes could be isolated as bilamellar membrane structures after cell disruption.
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