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Acrosome reaction on a Sea Urchin cell During fertilization, a sperm must first fuse with the plasma membrane and then penetrate the female egg in order to fertilize it. Fusing to the egg usually causes little problem, whereas penetrating through the egg's hard shell can present more of a problem to the sperm. Therefore sperm cells go through a process known as the acrosome reaction which is the reaction that occurs in the acrosome of the sperm as it approaches the egg. The acrosome is a cap-like structure over the anterior half of the sperm's head. As the sperm approaches the zona pellucida of the egg, which is necessary for initiating the acrosome reaction, the membrane surrounding the acrosome fuses with the plasma membrane of the sperm, exposing the contents of the acrosome. The contents include surface antigens and numerous enzymes which are responsible for breaking through the egg's tough coating and allowing fertilization to occur.
Contents
1 Variations among species o 1.1 Echinoderms o 1.2 Mammals 2 The process 3 In in vitro fertilization o 3.1 Assessment 4 See also
[edit] Echinoderms
In some lower animal species a protuberance (the acrosomal process) forms at the apex of the sperm head, supported by a core of actin microfilaments. The membrane at the tip of the acrosomal process fuses with the egg's plasma membrane. In some echinoderms, including starfish and sea urchins, a major portion of the exposed acrosomal content contains a protein that temporarily holds the sperm on the egg's surface.
[edit] Mammals
In mammals the acrosome reaction releases hyaluronidase and acrosin; their role in fertilization is not yet clear. The acrosomal reaction does not begin until the sperm comes into contact with the oocyte's jelly layer. After the jelly layer is penetrated, the acrosomal enzymes begin to dissolve and the actin filament comes into contact with the zona pellucida. Once the two meet, a calcium influx occurs, causing a signaling cascade. The cortical granules inside the oocyte then fuse to the outer membrane and a transient fast block reaction occurs. It also alters a patch of pre-existing sperm plasma membrane so that it can fuse with the egg plasma membrane. A sperm penetration assay includes an acrosome reaction test that assesses how well a sperm is able to perform during the fertilization process. Sperm that are unable to properly go through the acrosome reaction will not be able to fertilize an egg. However, this problem only occurs in about 5% of men that have the test done. This test is rather expensive and provides limited information on a man's fertility.[1] In other cases, such as in the wood mouse Apodemus sylvaticus, premature acrosome reactions have been found to cause increased motility in aggregates of spermatozoa promoting fertilization [2]
[edit] Assessment
Birefringence microscopy,[3] flow cytometry [4] or fluorescence microscopy can be used for assessing the shedding of the acrosome or "acrosome reaction" of a sperm sample. Flow cytometry and fluorescence microscopy are usually done after staining with a fluoresceinated lectin such as FITC-PNA, FITC-PSA, FITC-ConA, or fluoresceinated antibody such as FITC-CD46.[5] The antibodies/lectins have a high specificity for different parts of the acrosomal region, and will only bind to a specific site (acrosomal content/ inner/outer membrane). If bound to a fluorescent molecule, regions where these probes have bound can be visualised. Sperm cells with artificially induced acrosome reactions may serve as positive controls. For fluorescence microscopy a smear of washed sperm cells are made, airdried, permealized and then stained. Such a slide is then viewed under light of a wavelength that will cause the probe to fluoresce if it is bound to the acrosomal region. At least 200 cells are viewed in an arbitrary fashion and classified as either acrosome intact (fluorescing bright green) or acrosome reacted (no probe present, or only on the equatorial region). It is then expressed as a percentage of the counted cells. For assessment with flow cytometry the washed cells are incubated with the chosen probe, (possibly washed again) and then sampled in a flow cytometer. After gating the cell population according to forward- and side-scatter the resulting data can be analysed (E.g. mean fluorescences compared). With this technique a probe for viability, like propidium iodide (PI) could also be included in order to exclude dead cells from the acrosome assessment, since many sperm cells will spontaneously lose their acrosome when they die.