Action of phorbol myristate acetate (PMA) at fertilization of mouse oocytes in vitro

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 171-177
Author(s):  
Anna Niemierko ◽  
Aldona Komar
Keyword(s):  
Phorbol Ester ◽  
Phorbol Myristate Acetate ◽  
Cytochalasin B ◽  
Fertilization In Vitro ◽  
Myristate Acetate ◽  
Cleavage Division ◽  
Mouse Oocytes ◽  
Maturation Division ◽  
Timing Of Exposure

Phorbol ester (PMA) in concentration 5 and 10ng ml−1 blocks cytokinesis of the second maturation division in mouse oocytes. Karyokinesis is not impaired and digynic triploid oocytes are obtained which undergo first cleavage division. Effectiveness of blocking cytokinesis is dependent on the timing of exposure of oocytes to PMA action. When oocytes are subjected to PMA at the onset of the second maturation division only 14·5% of eggs are triploid. PMA present during fertilization in vitro (about 1 h exposure to PMA) induces triploidy in 40% eggs. Extending the time of exposure of oocytes to 2 h produces 76% tripronucleate eggs. Applicability of PMA is compared with the use of cytochalasin B to induce triploidy in the mouse.

scholarly journals Cytochalasin B-induced triploidy in mouse oocytes fertilized in vitro

Reproduction ◽  
1976 ◽  
Vol 48 (2) ◽  
pp. 279-284 ◽  
Author(s):  
A. Niemierko ◽  
A. Komar
Keyword(s):  
Cytochalasin B ◽  
Mouse Oocytes

Cytochalasin B-induced pseudo-cleavage of mouse oocytes in vitro. II. Studies of the mechanism and morphological consequences of pseudocleavage

1977 ◽  
Vol 26 (1) ◽  
pp. 323-337
Author(s):  
P.M. Wassarman ◽  
T.E. Ukena ◽  
W.J. Josefowicz ◽  
G.E. Letourneau ◽  
M.J. Karnovsky
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cytochalasin B ◽  
Oocyte Size ◽  
Contractile Ring ◽  
Subsequent Formation ◽  
Mouse Oocytes ◽  
Meiotic Competence ◽  
Scanning Electron

Mouse oocytes are induced by cytochalasin B to undergo ‘pseudocleavage’ in vitro into 2 compartments, only one of which possesses microvilli. It has been found that this particular response to cytochalasin B is related to oocyte size and, possibly, to the acquisition of meiotic competence by the oocyte during its growth phase. Certain of the morphological events which characterize pseudocleavage have been determined using transmission and scanning electron microscopy. These events include: (i) an initial withdrawal of microvilli from the surface of the oocyte, together with the concomitant disappearance of microfilaments normally associated with the microvilli; (ii) the subsequent formation of a pseudocleavage furrow and contractile ring; and (iii) the reappearance of microvilli and associated microfilaments in one of the two resulting oocyte compartments. These changes in surface architecture are reflected in the distribution of fluorescein-conjugated lectins bound to the oocyte surface during pseudocleavage.

Preimplantation development of gynogenetic diploid mouse embryos

Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 215-222
Author(s):  
Ewa Borsuk
Keyword(s):  
Cytochalasin B ◽  
Polar Body ◽  
Mouse Embryos ◽  
Fertilization In Vitro ◽  
Body Formation ◽  
Male Pronucleus ◽  
Step Procedure ◽  
Gynogenetic Diploid ◽  
Polar Body Formation

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.

“Autoregulation” of Human Neutrophil Activation In Vitro: Regulation of Phorbol Myristate Acetate-Induced Neutrophil Activation by Cell Density

1990 ◽  
Vol 47 (5) ◽  
pp. 457-474 ◽  
Author(s):  
Stephen P. Peters ◽  
Franklin Cerasoli ◽  
Kurt H. Albertine ◽  
Marlys H. Gee ◽  
David Berd ◽  
...  
Keyword(s):  
Cell Density ◽  
Phorbol Myristate Acetate ◽  
Human Neutrophil ◽  
Neutrophil Activation ◽  
Myristate Acetate

Suppressive action of cobalt on exocytosis and respiratory burst in neutrophils

1989 ◽  
Vol 257 (5) ◽  
pp. C859-C864 ◽  
Author(s):  
J. G. Elferink ◽  
M. Deierkauf
Keyword(s):  
Plasma Membrane ◽  
Phorbol Myristate Acetate ◽  
Respiratory Burst ◽  
Cytochalasin B ◽  
Superoxide Production ◽  
Co2 Activation ◽  
Chemotactic Peptide ◽  
Myristate Acetate ◽  
Intracellular Target ◽  
Suppressive Action

Activation of exocytosis and respiratory burst in rabbit neutrophils by the chemotactic peptide fMet-Leu-Phe is inhibited by Co2+. Inhibition is antagonized by extracellular Ca2+ and is dependent on the time of preincubation of cells with Co2+ before addition of activator. Co2+ inhibits the enhancement of 45Ca association that occurs during activation with fMet-Leu-Phe. Interference with Ca2+ translocation across the plasma membrane by Co2+ is probably not the cause of inhibition of neutrophil activation, because activation in the absence of extracellular Ca2+ is inhibited by Co2+. Activation of neutrophils by phorbol myristate acetate is inhibited at higher Co2+ concentrations than activation by fMet-Leu-Phe. Inhibition of the superoxide production by Co2+ occurs both in the presence or in the absence of cytochalasin B. Fluorescence of neutrophils loaded with quin2 is diminished by Co2+, indicating that Co2+ had entered into the cytoplasm. The results are compatible with the view that Co2+ inhibits exocytosis and respiratory burst in neutrophils by an interaction with a Ca2+-dependent intracellular target.

scholarly journals Involvement of the Ca2+-dependent phosphorylation of a 20 kDa protein in the proliferative effect of high-density lipoproteins (subclass 3) on the adenocarcinoma cell line A549

1995 ◽  
Vol 307 (2) ◽  
pp. 557-561 ◽  
Author(s):  
K A Tazi ◽  
M Bonnafous ◽  
G Favre ◽  
G Soula ◽  
F Le Gaillard
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Ester ◽  
Phorbol Myristate Acetate ◽  
High Density Lipoproteins ◽  
Myristate Acetate ◽  
Adenocarcinoma Cell Line ◽  
Adenocarcinoma Cell ◽  
Proliferative Effect ◽  
Kinase C

Previous studies from our laboratory demonstrated that high-density lipoproteins (subclass 3; HDL3) bind to sites specific for apolipoprotein AI on the human adenocarcinoma cell line A549 and that HDL3 binding promotes a mitogenic effect [Favre, Tazi, Le Gaillard, Bennis, Hachem and Soula (1993) J. Lipid Res. 34, 1093-1106]. In the present study, we have examined the cell proteins that showed modified phosphorylation after binding of HDL3 to specific sites, and the roles of Ca2+ and protein kinase C. Native HDL3 (but not tetranitromethane-modified HDL3) and Ca2+ ionophore A23187 strongly enhanced the phosphorylation of a 20 kDa protein (x 3.6) and, to a lower extent, the phosphorylation of 24 and 28 kDa proteins (x 2.2 and 2.6 respectively). The two effectors were equally able to stimulate cell growth. Down-regulation of protein kinase C by a 24 h incubation of cells with phorbol myristate acetate prevented the effects of HDL3 on the phosphorylation of 24 and 28 kDa proteins. However, the extent of phosphorylation of the 20 kDa protein was not affected. In contrast, activation of protein kinase C by a short incubation with phorbol myristate acetate resulted in inhibition of proliferation and an increase in 24 and 28 kDa (but not 20 kDa) protein phosphorylation. These results suggest that HDL3 putative receptors exert their proliferative effect on A549 cells through activation of a Ca(2+)-dependent protein kinase. This kinase activity is not modulated by phorbol ester and thus may be a calmodulin kinase or an isoenzyme of protein kinase C that is independent of phorbol ester. It allows a subsequent 20 kDa protein to be phosphorylated.

Fertilization in vitro of frozen mouse oocytes

Cryobiology ◽  
1978 ◽  
Vol 15 (6) ◽  
pp. 689 ◽  
Author(s):  
M. Kasai ◽  
K. Niwa ◽  
A. Iritani
Keyword(s):  
Fertilization In Vitro ◽  
Mouse Oocytes
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