Acrosome reaction
Encyclopedia
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.
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.
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.
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
of the fallopian tube
(site of fertilization) when the sperm penetrates the secondary oocyte. A few events precede the actual acrosome reaction. The sperm cell acquires a "hyperactive motility pattern" by which its flagellum produces vigorous whip-like movements that propel the sperm through the cervical canal and uterine cavity, until it reaches the isthmus of the fallopian tube. The sperm approaches the ovum in the ampulla of the fallopian tube with the help of various mechanisms, including chemotaxis
. Glycoproteins on the outer surface of the sperm then bind with glycoproteins on the zona pellucida
of the ovum.
The first stage is the penetration of corona radiata, by releasing hyaluronidase from the acrosome to digest cumulus cells surrounding the oocyte and exposing acrosin attached to the inner membrane of the sperm. The cumulus cells are embedded in a gel-like substance made primarily of hyaluronic acid, and developed in the ovary with the egg and support it as it grows. After reaching the Zona Pellucida
the actual acrosome reaction begins.
Acrosin digests the zona pellucida and membrane of the oocyte. Part of the sperm's cell membrane then fuses with the egg cell's membrane, and the contents of the head sink into the egg. In the mouse it has been demonstarated that ZP3, one of the proteins that make up the zona pellucida, binds to a partner molecule (to the β1,4-galactosyl transferase receptors) on the sperm. This lock-and-key type mechanism is species-specific and prevents the sperm and egg of different species from fusing. There is some evidence that this binding is what triggers the acrosome to release the enzymes that allow the sperm to fuse with the egg. It is likely that a similar mechanism occurs in other mammals, but the diversity of zona proteins across species means that the relevant protein and receptor may differ.
Upon penetration, if all is occurring normally, the process of egg-activation occurs and the oocyte is said to have become activated. This is thought to be induced by a specific protein phospholipase c zeta. It undergoes its secondary meiotic division, and the two haploid nuclei (paternal and maternal) fuse to form a zygote
. In order to prevent polyspermy
and minimise the possibility of producing a triploid zygote, several changes to the egg's cell membranes renders them impenetrable shortly after the first sperm enters the egg.
The aforementioned process describes the physiologically relevant events. One should however bear in mind that a certain percentage of sperm cells will undergo a spontaneous acrosome reaction without the presence of the ovum. Those cells are not able to fertilise the egg, even if they do reach it later. Other cells will spontaneously shed their acrosome during the process of apoptosis/necrosis.
(ICSI) for IVF, the implantation rate is higher in oocytes injected with spermatozoa that have undergone acrosome reaction (~40%) vs. those injected with nonreacted spermatozoa (~10%). The implantation rate is ~25% in when injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle follows the same trend.
The acrosome reaction can can be stimulated in vitro by substances a sperm cell may encounter naturally such as progesterone
or follicular fluid
, as well as the more commonly used calcium ionophore
A23187.
microscopy, flow cytometry
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. 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.
Sperm
The term sperm is derived from the Greek word sperma and refers to the male reproductive cells. In the types of sexual reproduction known as anisogamy and oogamy, there is a marked difference in the size of the gametes with the smaller one being termed the "male" or sperm cell...
must first fuse with the plasma membrane and then penetrate the female egg
Ovum
An ovum is a haploid female reproductive cell or gamete. Both animals and embryophytes have ova. The term ovule is used for the young ovum of an animal, as well as the plant structure that carries the female gametophyte and egg cell and develops into a seed after fertilization...
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
Acrosome
The acrosome is an organelle that develops over the anterior half of the head in the spermatozoa of many animals. It is a cap-like structure derived from the Golgi apparatus. Acrosome formation is completed during testicular maturation. In Eutherian mammals the acrosome contains digestive enzymes...
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
Zona pellucida
The zona pellucida is a glycoprotein membrane surrounding the plasma membrane of an oocyte. It is a vital constitutive part of the latter, external but of essential importance to it...
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.
Variations among species
There are considerable species variations in the morphology and consequences of the acrosome reaction. In several species the trigger for the acrosome reaction has been identified in a layer that surrounds the egg.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.
Mammals
In mammals the acrosome reaction releases hyaluronidaseHyaluronidase
The hyaluronidases are a family of enzymes that degrade hyaluronic acid.In humans, there are six associated genes, including HYAL1, HYAL2, HYAL3, and PH-20/SPAM1.-Use as a drug:...
and acrosin
Acrosin
Acrosin is a digestive enzyme that acts as a protease. In humans, acrosin is encoded by the ACR gene. Acrosin is released from the acrosome of spermatozoa as a consequence of the acrosome reaction. It aids in the penetration of the Zona Pellucida....
; 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.
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
The process
The acrosomal reaction normally takes place in the ampullaAmpulla of uterine tube
The ampulla is the second portion of the uterine tube. It is an intermediate dilated portion, which curves over the ovary.It is the most common site of human fertilization.The word ampulla is from Latin for flask....
of the fallopian tube
Fallopian tube
The Fallopian tubes, also known as oviducts, uterine tubes, and salpinges are two very fine tubes lined with ciliated epithelia, leading from the ovaries of female mammals into the uterus, via the utero-tubal junction...
(site of fertilization) when the sperm penetrates the secondary oocyte. A few events precede the actual acrosome reaction. The sperm cell acquires a "hyperactive motility pattern" by which its flagellum produces vigorous whip-like movements that propel the sperm through the cervical canal and uterine cavity, until it reaches the isthmus of the fallopian tube. The sperm approaches the ovum in the ampulla of the fallopian tube with the help of various mechanisms, including chemotaxis
Chemotaxis
Chemotaxis is the phenomenon in which somatic cells, bacteria, and other single-cell or multicellular organisms direct their movements according to certain chemicals in their environment. This is important for bacteria to find food by swimming towards the highest concentration of food molecules,...
. Glycoproteins on the outer surface of the sperm then bind with glycoproteins on the zona pellucida
Zona pellucida
The zona pellucida is a glycoprotein membrane surrounding the plasma membrane of an oocyte. It is a vital constitutive part of the latter, external but of essential importance to it...
of the ovum.
The first stage is the penetration of corona radiata, by releasing hyaluronidase from the acrosome to digest cumulus cells surrounding the oocyte and exposing acrosin attached to the inner membrane of the sperm. The cumulus cells are embedded in a gel-like substance made primarily of hyaluronic acid, and developed in the ovary with the egg and support it as it grows. After reaching the Zona Pellucida
Zona pellucida
The zona pellucida is a glycoprotein membrane surrounding the plasma membrane of an oocyte. It is a vital constitutive part of the latter, external but of essential importance to it...
the actual acrosome reaction begins.
Acrosin digests the zona pellucida and membrane of the oocyte. Part of the sperm's cell membrane then fuses with the egg cell's membrane, and the contents of the head sink into the egg. In the mouse it has been demonstarated that ZP3, one of the proteins that make up the zona pellucida, binds to a partner molecule (to the β1,4-galactosyl transferase receptors) on the sperm. This lock-and-key type mechanism is species-specific and prevents the sperm and egg of different species from fusing. There is some evidence that this binding is what triggers the acrosome to release the enzymes that allow the sperm to fuse with the egg. It is likely that a similar mechanism occurs in other mammals, but the diversity of zona proteins across species means that the relevant protein and receptor may differ.
Upon penetration, if all is occurring normally, the process of egg-activation occurs and the oocyte is said to have become activated. This is thought to be induced by a specific protein phospholipase c zeta. It undergoes its secondary meiotic division, and the two haploid nuclei (paternal and maternal) fuse to form a zygote
Zygote
A zygote , or zygocyte, is the initial cell formed when two gamete cells are joined by means of sexual reproduction. In multicellular organisms, it is the earliest developmental stage of the embryo...
. In order to prevent polyspermy
Polyspermy
In biology, polyspermy describes an egg that has been fertilized by more than one sperm. Diploid organisms normally contain two copies of each chromosome, one from each parent. The cell resulting from polyspermy, on the other hand, contains three or more copies of each chromosome -- one from the...
and minimise the possibility of producing a triploid zygote, several changes to the egg's cell membranes renders them impenetrable shortly after the first sperm enters the egg.
The aforementioned process describes the physiologically relevant events. One should however bear in mind that a certain percentage of sperm cells will undergo a spontaneous acrosome reaction without the presence of the ovum. Those cells are not able to fertilise the egg, even if they do reach it later. Other cells will spontaneously shed their acrosome during the process of apoptosis/necrosis.
In in vitro fertilization
When using intracytoplasmic sperm injectionIntracytoplasmic sperm injection
Intracytoplasmic sperm injection is an in vitro fertilization procedure in which a single sperm is injected directly into an egg.-Indications:...
(ICSI) for IVF, the implantation rate is higher in oocytes injected with spermatozoa that have undergone acrosome reaction (~40%) vs. those injected with nonreacted spermatozoa (~10%). The implantation rate is ~25% in when injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle follows the same trend.
The acrosome reaction can can be stimulated in vitro by substances a sperm cell may encounter naturally such as progesterone
Progesterone
Progesterone also known as P4 is a C-21 steroid hormone involved in the female menstrual cycle, pregnancy and embryogenesis of humans and other species...
or follicular fluid
Follicular fluid
Follicular fluid is a liquid which fills the follicular antrum and surrounds the ovum in an ovarian follicle. This fluid is rich in hyaluronic acid.-External links:* *...
, as well as the more commonly used calcium ionophore
Ionophore
An ionophore is a lipid-soluble molecule usually synthesized by microorganisms to transport ions across the lipid bilayer of the cell membrane...
A23187.
Assessment
BirefringenceBirefringence
Birefringence, or double refraction, is the decomposition of a ray of light into two rays when it passes through certain anisotropic materials, such as crystals of calcite or boron nitride. The effect was first described by the Danish scientist Rasmus Bartholin in 1669, who saw it in calcite...
microscopy, flow cytometry
Flow cytometry
Flow cytometry is a technique for counting and examining microscopic particles, such as cells and chromosomes, by suspending them in a stream of fluid and passing them by an electronic detection apparatus. It allows simultaneous multiparametric analysis of the physical and/or chemical...
or fluorescence microscopy
Fluorescence microscope
A fluorescence microscope is an optical microscope used to study properties of organic or inorganic substances using the phenomena of fluorescence and phosphorescence instead of, or in addition to, reflection and absorption...
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
Lectin
Lectins are sugar-binding proteins that are highly specific for their sugar moieties. They play a role in biological recognition phenomena involving cells and proteins. For example, some viruses use lectins to attach themselves to the cells of the host organism during infection...
such as FITC-PNA, FITC-PSA, FITC-ConA, or fluoresceinated antibody such as FITC-CD46. 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.