Early mouse development - lecture 11 Flashcards Preview

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1

What are the first four stages of a mouse development

- blastomeres , blastula , morula , blastocyst

2

what is rotational cleavage?

Can be an odd number of blastomeres at certain times

3

When does hatching occur?

At the blastocyst stage , secretes enzymes which break down the zona pellucida

4

What is the blastocyst made up of?

Inner cell mass surrounded by trophoblast cells and contains a fluid filled cavity called a blastocoel

5

What happens prior to implantation, to inner cell mass?

Formation of two layers; the epiblast and the hypoblast (primitive endoderm)

6

What is the role of the hypoblast (primitive endoderm)?

lines the blastocoel forming the yolk sac.

7

What is the role of the epiblast?

forms the amniotic cavity

8

What is the role of the trophoblast?

continue to divide to invade the uterus forming the syncytiotrophoblast

9

What does the posterior node of the epiblast signal?

signals for a primitive streak to form which then forms a groove

10

What happens to the epiblast cells during gastrulation?

Migrate through the primitive groove , invading endoderm cells.

11

What happens after this process of ingression?

formation of 3 germ layers, ectoderm, mesoder and endoderm. The cells which have moved through the node , will form the notochord

12

What is a notochord?

a cartilaginous skeletal rod supporting the body

13

What does the ectoderm develop into?

- epidermis
- brain , CNS

14

What does the mesoderm develop into?

- muscle , connective tissues , bone
- kidney and gonads
- tissues of the heart

15

What does the endoderm develop into?

- epithelium lining of the gastrointestinal tract
- epithelium lining of respiratory tract
- liver, lungs , pancreas

16

How to add a gene into a mouse

- injection of DNA into pronuclei by micropipette after fertilisation
- DNA integrates into the chromosome
- implant egg in to surrogate oviduct
- determine if gene is taken up via GFP. or screened by PCR reaction

17

What is the procedure to make a targeted knockout mouse?

- obtain ES cells
- Make a targeting construct
- Genetically modify ES cells , by inserting the targeting vector
- inject new ES cells into blastocyst
- implant recombinant embryo into surrogate
- breed pups

18

Who won the Nobel prize for targeted mouse knockouts?

- Sir Martin Evans
- Mario Capecchi
- Oliver Smithies

19

What happens when you knock out the Pdx1 gene?

no pancreas development

20

How do you make a targeting construct?

- Linearized DNA construct is added to ES cell with an electric current.
- DNA construct moves into the nucleus where homologous recombination can take place.
- Recombination can occur and gene of interest is knocked out