Second polar body formation in rat ovarian oocytes matured in vitro

Author(s):  
G.H. Zeilmaker ◽  
C.M.P.M. Verhamme
Development ◽  
1974 ◽  
Vol 31 (2) ◽  
pp. 497-512
Author(s):  
C. F. Graham ◽  
Z. A. Deussen

Unfertilized mouse eggs were activated in vitro with hyaluronidase. Subsequently they were exposed to culture medium at different osmolarities. In full strength White's culture medium they tended to form one pronucleus and a second polar body. The majority of these eggs were haploid. In 4/5 and 3/5 dilutions of this medium, the second polar body formation was suppressed and eggs tended to form with one or two pronuclei. Those with one pronucleus were diploid and those with two pronuclei could either form a diploid or form a haploid mosaic. Old eggs tended to immediately cleave and form haploid mosaics. DNA synthesis was studied in activated eggs using tritiated thymidine and autoradiography. DNA synthesis occurred at a similar time in fertilized and activated eggs.


Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 215-222
Author(s):  
Ewa Borsuk

Diploid gynogenetic mouse embryos were produced in a three-step procedure: fertilization in vitro, suppression of the 2nd polar body formation by Cytochalasin B, and microsurgical removal of the male pronucleus. The operated eggs were transplanted to the oviduct of recipient females for 72 or 96 h. The overall recovery rate was 73%, but compacted morulae and blastocysts constituted only 28·6% of transplanted eggs. After 72 h blastocysts were rare (3·5%) but 24 h later their incidence increased to 21·2%. In eggs homozygous for T6 chromosome it was possible to prove karyologically that the male pronucleus was effectively removed and that the diploid genome was of purely maternal origin.


1935 ◽  
Vol s2-77 (308) ◽  
pp. 585-604
Author(s):  
MARGOT E. METEALFE

1. The somatic cells in both sexes of Phytophaga destructor Say contain four pairs of V-shaped chromosomes, the sex-group being indistinguishable in size or form. 2. The germ-cells in both sexes contain eight pairs of chromosomes. 3. The maturation of the egg follows the normal course of development, eight bivalents being formed. After polar body formation the female pronucleus has eight chromosomes. The polar bodies are never extruded from the egg. 4. Spermatogenesis is a complicated process, the details of which have not been satisfactorily determined. The growth stage appears to take place before the last spermatogonial division. No pairing of chromosome has been observed, and apparently no metaphase plate is formed at meiosis. Eeduction is effected by the expulsion of two buds each containing four chromosomes. Thus only one sperm is produced from each spermatocyte. 5. One or more sperms may enter the egg at fertilization. 6. The germ-line is differentiated from the soma at the eightcell stage. 7. At the fifth cleavage the somatic nuclei eliminate half their number of chromosomes, and are left with eight chromosomes. 8. Migration of the germ nuclei takes place at the sixteencell stage. 9. The relation of the chromosome numbers in the somatic and germ lines is discussed.


Development ◽  
1974 ◽  
Vol 31 (2) ◽  
pp. 513-526
Author(s):  
M. H. Kaufman ◽  
M. A. H. Surani

Eggs from (C57B1 × A2G)F1 mice were activated by treatment with hyaluronidase, which removed the follicle cells, and cultured in vitro. Observations were made 6–8 h after hyaluronidase treatment to determine the frequency of activation and the types of parthenogenones induced. Cumulus-free eggs resulting from hyaluronidase treatment were incubated for 2¼ h in culture media of various osmolarities. The frequency of activation was found to be dependent on the postovulatory age of oocytes, while the types of parthenogenones induced were dependent on the osmolarity of the in vitro culture medium and their postovulatory age. Culture in low osmolar medium suppressed the extrusion of the second polar body (2PB). This decreased the incidence of haploid eggs with a single pronucleus and 2PB and immediately cleaved eggs from 97·5% to 42·3% of the activated population. Where 2PB extrusion had been suppressed, 97·4% of parthenogenones contained two haploid pronuclei. Very few were observed with a single and presumably diploid pronucleus. Serial observations from 11 to 18 h after hyaluronidase treatment were made on populations of activated eggs as they entered the first cleavage mitosis after 2¼ h incubation in medium either of normal (0·287 osmol) or low (0·168 osmol) osmolarity. A delay in the time of entry into the first cleavage mitosis similar to the duration of incubation in low osmolar medium was observed. Further, eggs were incubated in control and low osmolar culture media containing uniformly labelled [U-14C]amino acid mixture to examine the extent of protein synthesis in recently activated eggs subjected to these culture conditions. An hypothesis is presented to explain the effect of incubation in low osmolar culture medium in delaying the first cleavage mitosis.


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