Final Exam - Mouse Development Flashcards
(31 cards)
Fertilization
- the fusion of sperm and egg to form the zygote
- occurs in the oviduct
Cleavage
- a series of slow (in mouse every 12 hours) cell divisions without cell growth that occurs after fertilization and divides the embryo up into a number of small cells called blastomeres
- two-cell, four-cell, eight-cell, compacted morula, blastocyst
Blastomere compaction
- difference from frogs
- happens at the 8-cell stage
- asymmetry within embryo is established
- cells become polarized
- cells have smooth membranes inside the embryo
- outer membranes become covered with microvilli to increase surface area of cell-cell contact
Morula
- the very early stage in a mammalian embryo when cleavage has resulted in a solid ball of cells
- forms at the 8-cell stage (characterized by blastomere compaction)
- has 32 cells with two distinct groups: trophectoderm and inner cell mass (ICM)
Trophectoderm
- a group of outer cells of the morula
- the outer layer of cells (20 cells) of the early mammalian embryo
- it gives rise to extra-embryonic structures and tissues such as the placenta
Inner cell mass (ICM)
- a group of inner cells of the morula
- a discrete mass of cells in the blastocyst of the early mammalian embryo which is derived from the inner cells of the morula, and which will give rise to the embryo proper and some extra-embryonic membranes
Blastocyst
- the stage of a mammalian embryo that corresponds in form to the blastula stage of other animal embryos , and is the stage at which the embryo implants in the uterine wall
- is formed as trophectoderm pumps in fluid to create a blastocoel (inner cell mass becomes created on one end)
Primitive endoderm
-in mammalian embryos, that part of the inner cell mass that contributes to extra-embryonic membranes
Primitive endoderm
- in mammalian embryos, that part of the inner cell mass that contributes to extra-embryonic membranes
- later becomes parietal endoderm, a sticky layer of protective cells
Epiblast/primitive ectoderm
- in mouse and chick embryos, a group of cells within the blastocyst or blastoderm, respectively, that gives rise to the embryo proper
- in the mouse, it develops from cells of the inner cell mass
Placenta
- a structure that forms in the uterine wall at which the blood systems of mother and embryo form an interface with each other
- mammalian embryos (with the exception of the monotremes like egg-laying platypus and echidnas) are nourished by the mother by the passage of nutrients through the placenta
- formed by trophectoderm (its normal fate), which consists in part of trophoblast giant cells
Parietal endoderm
- starts out as primitive endoderm which then becomes the parietal endoderm
- is a sticky layer of protective cells
Visceral endoderm
- tissue derived from the primitive endoderm that develops on the surface of the egg cylinder in the mammalian blastocyst
- Lies next to epiblast (embryo)
- Has an early developmental function
Trophoblast giant cells
- found in placenta
- Are specialized cells that endoreduplicate (many copies of its genome in a single nucleus, make more copies of DNA to make more genes) = production machines
- Invade the uterine wall and mediate the interface between vascular systems of the mother and embryo
Primitive streak
- gastrulation initiates with the formation of the primitive streak
- the site of gastrulation in avian and mammalian embryos and the forerunner of the antero-posterior axis
- it is a strip of ingressing cells that extends into the epiblast from the posterior margin
- epiblast cells move through the streak into the interior of the embryo to form mesoderm and endoderm
Mouse (Mus musculus) embryos
- Embryos are very small (approx. 1/10 of a mm)
- Implantation, placenta development
Embryonic stem cells (ES cells)
- pluripotent cells derived from the inner cell mass of a mammalian embryo, most commonly mouse embryo, that can be indefinitely maintained in culture
- when injected into another blastocyst, they combine with the inner cell mass and can potentially contribute to all the tissues of the embryo
- can stay in the stem cell fate, grow and replicate
- can be used to genetically manipulate transgenic (chimeric) animals
- The modified ES cells can be injected back into the blastocyst and can contribute to the inner cell mass
- This creates chimeric embryos (a mix of donor and host tissue)
- If these cells contribute to the germ line they will be inherited by the next generation
- These can be called clones
Transgenic/chimeric animals
- an organism whose genetic make-up is the result of the deliberate introduction of new DNA
- ex) new genes, genes with specific mutations, DNAs that cause the inactivation of specific genes
- term is also used to describe the various genetic engineering techniques that can be used to accomplish this
- becomes a clone
- Can be manipulated (such that their genomes can be modified) so new genes are introduced=transgenes
- Genes could be manipulated such that genes could be knocked out
- chimeric embryo=a mix of donor and host tissue, a clone
Transgenes
- trans=coming from an external source
- a gene, sometimes from another organism, that has been introduced into a cell or organism by genetic engineering techniques
Genetic “knock-outs”
- the complete and permanent inactivation or deletion of a particular gene in an organism by means of genetic manipulation
- genes can be deleted and/or removed
What are the stages undergone by the early embryo?
1) cleavage
2) blastomere compaction
3) morula (8-cell stage)
4) morula (32-cell stage with trophectoderm and inner cell mass)
5) blastocyst
Drug-resistance selection
-Drug-resistance selection is used for selection of ES (embryonic stem) cells that have been modified
Cre recombinase/loxP
- Cre recombination through loxP sites
- Genes that are to be deleted are flanked by loxP sites
- These are acted upon by cre-recombinase which expressed under specific enhancers/promoters
- This was used to knockout genes
- Promoters can be tissue and time specific such that gene knockout can be targeted to specific time in development
Crisper/Cas9
- Cas 9 is a nuclease (endonuclease) derived from bacteria
- These nucleases can cleave DNA
- Guide RNA encoded by a Crisper array
- These target the nuclease to specific sites
- Cleavage is then repaired by the cell’s normal DNA repair mechanisms
- This is used by researchers to incorporate custom DNA sequences into the genome