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Developmental Biology: LS 311/ 411 External Fertilization
Developmental Biology: LS 311/ 411 External Fertilization
Lampbrush chromosomes
(ref. gilbert images)
They have very active chromosomes that synthesize large amounts of RNA (mRNA, tRNA, rRNA).
Meroistic Oogenesis
A type of oogenesis where cytoplasmic connection remains between the cells produced by the oogonium. Unlike amphibians, the oocyte of insects dont pass through transcriptionally active stage. Rather RNA synthesis is confined to other supporting cells and transported to the oocyte.
In Drosophila melanogaster, the oogonia are called cytoblasts, and they undergo an unusual specialization They undergo multiple mitotic divisions, but fail to undergo cytokinesis (cell division). Thus, they all remain connected to the original cell as cytocytes One of the cytocysts becomes the oocyte
The other 15 become nurse cells. They make large amounts of RNA and nutrients but they send it all to the oocyte. This allows the oocyte to accumulate massive amounts of cytoplasm to support development (15 nuclei instead of 1).
Gilberts Book
Structure of Spermatozoon
Head Acrosome Nucleus Middle Piece -Mitochondria Flagellum Axoneme
One of the most fundamental questions of external fertilization is how do sperm locate eggs? Chemo-attractant source mediates attraction to the egg How can sperm be prevented from trying to fertilize eggs of another species? Species-specific recognition
Sperm Attraction
Species specific sperm attraction has been documented in some invertebrates In Echinoderms a small peptide, Resact, secreted from the egg jelly attracts the sperms
Increase in cGMP and Ca2+ activates mitochondrial ATP generation and dynein ATPase stimulating flagellar movement in sperm.
1. Acrosome Reaction
Acrosome Reaction
Acrosomal reaction is initiated by the specific sugars present in the egg jelly.
Acrosome reaction has two components: 1. Exocytosis of the acrosomal contents 2. Extension of acrosomal process
Elevation of intracellular Ca2+ a. Triggers fusion of acrosomal membrane with adjacent sperm membrane b. Activates acrosomal process formation by activating Rho B that stimulates polymerization of globular actin to actin filaments
Acrosomal reaction also involves the extension of acrosomal process formed by the polymerization of the globular actin.
Species-Specific Recognition
Sperm attraction is species specific Acrosomal reaction is species specific Acrosomal protein like Bindin, mediates the species specific interaction of sperm with the vitelline envelope.
Species-specific bindin isolated from the acrosomes of Strongylocentrotus purpuratus binds to its own dejellied eggs, but not to those of Arbacia
punctulata.
Summary
1. Egg secrete soluble molecules for chemoattraction of sperms. 2. The exocytosis of the acrosomal vesicle to release its enzymes 3. Binding of sperm to the external envelope of the egg 4. Passage of sperm through the extracellular envelope 5. Fusion of egg and sperm cell membranes
Early responses:
Fast block initiated by influx of Na+ Slow block initiated by intracellular release of Ca 2+ leading to cortical granule reaction Ca 2+ release activates a whole of series of metabolic reactions a. Affecting lipid biosynthesis (new membranes are formed) b. Leads to respiratory burst crosslinking of fertilization envelope (peroxidase) c. Antioxidants like glutathione and ovothiols are generated that prevent damage to DNA
Late Responses:
Activation of DNA & protein synthesis. Protein synthesis is dependent on the stored messages in the egg and mediated probably by release of inhibitors from the mRNAs. In sea urchin egg translational initiation factor eIF4E in inhibited Post fertilization, the inhibitor is degraded and mRNAs are freed for translation. Cyclin B is one the cell cycle regulator is synthesized, creates Mitosis promoting factor (MPF), required to initiated cell division.
Ref. gilbert.
Mammalian fertilization
(Internal Fertilization)
Capacitation
Cholesterol efflux from the membrane Particular proteins or carbohydrates that block the recognition sites for sperm and zona interaction are removed Membrane potential of the sperm changes probably facilitating the acrosomal reaction Protein phosphorylation events that mediate sperm and zona pellucida interaction Changes in outer acrosomal membrane that prepares the fusion of sperm to egg.
Proposed model for the involvement of tyrosine-phosphorylated molecular chaperones in the acquisition of mammalian sperm fertilizing ability
Intial tethering of sperm and egg accomplished SED1 sperm cell surface adhesion protein binds to zona protein complex GalT on sperm surface recognizes Nacetylglucosamine moieties on ZP3
Gamete Fusion
Unlike the sea urchins, mammalian sperm contacts the egg at it sides and not at the tip It is believed that CD9 (in the egg) and immunoglobilin like protein Izumo (in sperm) mediate fusion of sperm to egg membrane.
Prevention of Polyspermy
Polyspermy is not a problem when the fertilization is internal In mammals the cortical granule reaction doesnt produce a fertilization envelope, but releases enzymes (like Nacetylglucosaminidase) that modify the Zona pellucida so that they no longer bind sperm.
Summary of events leading to the fusion of egg and sperm plasma membranes
External Fertilization Internal Fertilization
A. punctulata eggs release resact (CVTGAPGCVGGGRL), Speract (GFDLNGGGVG) is a related SAP that is released by eggs of another sea urchin, Strongylocentrotus purpuratus
Cleavage is a series of extremely rapid mitotic divisions wherein the enormous volume of zygote cytoplasm is divided into numerous smaller cells. These cells are called blastomeres, and by the end of cleavage, they generally form a sphere known as a blastula.
2. After the rate of mitotic division has slowed down, the blastomeres undergo dramatic movements wherein they change their positions relative to one another. This series of extensive cell rearrangements is called gastrulation, and the embryo is said to be in the gastrula stage. 3. As a result of gastrulation, the embryo contains three germ layers: the ectoderm, the endoderm, and the mesoderm.