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Superovulation treatment of the female with hormones so that more eggs are ovulated than normal
Superovulation alter normal level of FSH Follicle Stimulating HormoneTypical results are ovulation of 10 or more eggs
Superovulation recipient animals must be closely synchronized with the donor cow. Must be at the same stage in their heat cycle
EmbryoTransfer provides opportunity to introduce genetic material into populations of livestock while greatly reducing risk of transmission of diseases
ET Process super ovulate donor cow synchronize recipientsbreed donor cow (A.I.) when in heat
ET Process flush - remove fertilized eggs from donor before 7 days gestation plant embryo in the recipient animal
Transfer to recip. Non surgical commondeposit embryo in uterus through cervix with AI straw and gun
non surgical usually 70-80% success rate
Surgical incision in recip through flanktip of uterine horn is exposed cut open and embryo is deposited
Surgical recip is sewn shut less than 60-65% success rate.
ET now allows for shipping of 500 registered holsteins in four sealed flasks in one seat on an airplane
Splitting Embryos remove embryo from cow 7 days after conceptioncollect unfertilized egg and cut into zona pellucida
Splitting Embryos remove contents of unfertilized egg leaving the zona intact similarly, cut into zona around embryo
Splitting Embryos divide cells in embryo in half using a micro surgical blade remove 1/2 of the cells and transfer them into empty zona
Splitting Embryos seal zona with microjolteach 1/2 put into recipient animals
Cloning 6 days after conception, 32 cell embryo is removed from donor cow recipient egg cell is taken from a slaughterhouse
Cloning recipient cells DNA is removedone of the 32 cells is removed from the embryo
Cloning the single cell is inserted into recipient eggrecipient cell is given a microjolt - fuses grafted DNA to plasma membrane
Cloning desired embryo will replace cells that are removed providing a continuing supply of DNA to be inserted into more recipient cells
Cloning resulting offspring are cloned identical siblings
Sexing Embryos usually done by determining if embryo is a malemales have Y chromosome H-Y antigen can be detected in male cells
Detection of Y break open cells and stain chromosomes to identify presence of Y chromosome Karyotyping must examine at least 10 cells
Detection of Y 70% accurate use Y specific probe for IDY specific probe attaches to Y chromosome
Detection of Y can be detected by radioactivity or special light85% accurate
H-Y Antigen make an antibody by injecting H-Y antigen into miceattach dye onto the antibody for easy detection of male embryo
H-Y Antigen add antibody to embryo and if embryo is male it will glowif it is female it will remain dark
Advantages does not kill cell no damage to embryofast - can be few hours or less than one hour old
Disadvantages need high skilled technician 80% accurate
Freezing Embryos cooling embryos -5C or 40 Fcan be stored several days freezing -196 C or -320 F must cool before freezing
Freezing Embryos slowly cooled to -25C at a rate of .5C per minute and then plunged into liquid Nitrogen
Freezing cryoprotectants are used to lower the freezing point so removal of water can be increased
Freezing Cryoprotectant used in cattle - glycerolalso help prevent cell membrane from becoming brittle dilutes salt in cell
Thawing 500 C per minute complete is about 20 secondsglycerol is removed and embryo is placed in successive solutions
Thawing glycerol is removed and embryo is placed in successive solutions to re hydrate
Thawing if done too slowly embryo will burst because large ice crystals will form 70-80% success rate
Animals Genetic Transformation two tools embryo micromanipultionrecombinant DNA technology
Genetic Trans. Occurs when new genetic info is introduced into animals genome
Methods ES - Embryonic Stem EC - Embryonic Carcinoma cells
ES or EC cells harboring the foreign gene are incorporated into blastocyst resulting in a mosaic animal
EC potential for tumor formation
Retrovirus infection retro. Carrying desired gene is used to transformhost close to 100% efficient
Microinjection of several hundred copies of DNA are injected into male pronucleus of one or two cell embryos
Microinjection inefficient mice - 10-15% are bornonly 25% express new trait
Screening is it transgenic? Incorporation of gene expression of genegermline incorporation
Agricultural Use rabbit sheep pig cow
The End !! :)
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