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Fig. 5. Rsk is necessary for preventing parthenogenetic activation after meiosis
I in starfish oocytes. (A,B) Emission of only one polar body
from oocytes lacking Rsk activity, followed by parthenogenesis. Immature
oocytes were injected with 3 ng of control IgG or the neutralizing anti-Rsk
antibody 2, and then treated with 1-MeAde. These oocytes were either fixed at
2.5 hours after 1-MeAde addition, followed by DNA staining with DAPI (A), or
further incubated in the absence of insemination (B). The numbers of eggs
examined are indicated in parentheses. Arrowheads indicate polar bodies and
arrows indicate nucleus (A). Photographs of blastomere and gastrula were taken
7 and 22 hours, respectively, after 1-MeAde addition (B). (C) Dynamics
of H1 kinase activity in oocytes lacking Rsk activity. Extracts were prepared
at 15-minute intervals after 1-MeAde addition from oocytes that had been
injected with 3 ng of control IgG or the neutralizing anti-Rsk antibody 2.
Histone H1 kinase activity was detected on an autoradiogram (upper), which was
then quantified (lower). (D) Dynamics of phosphorylation states of
meiotic cell cycle regulators in oocytes lacking Rsk activity. Lysates from
oocytes, that had been injected with control IgG or the neutralizing anti-Rsk
antibody 2, were prepared at 15-minute intervals after 1-MeAde addition, and
immunoblotted with anti-Rsk serum 2, anti-Myt1, anti-Cdc25, anti-MAPK,
anti-active MAPK, anti-phospho Tyr-Cdc2 antibodies.
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