Polo-like kinase-1 is a pivotal regulator of microtubule assembly during mouse oocyte meiotic maturation, fertilization, and early embryonic mitosis.
Polo-like kinases (Plks) are a family of serine/threonine protein kinases that have been activated through phosphorylation. The activity of these kinases has been shown to be required for regulating multiple stages of mitotic progression in somatic cells. In this experiment, the changes in Plk1 expression were detected in mouse oocytes through Western blotting. The subcellular localization of Plk1 during oocyte meiotic maturation, fertilization, and early cleavage as well as after antibody microinjection or microtubule assembly disturbance was studied by confocal microscopy. The quantity of Plk1 protein remained stable during meiotic maturation and decreased gradually after fertilization. Plk1 was localized to the spindle poles of both meiotic and mitotic spindles at the early M phase and then translocated to the middle region. At anaphase and telophase, Plk1 was concentrated at the midbody of cytoplasmic cleavages. Plk1 was concentrated between the male and female pronuclei after fertilization. Plk1 disappeared at the spindle region when microtubule formation was inhibited by colchicine or staurosporine, while it was concentrated as several dots in the cytoplasm after taxol treatment. Plk1 antibody injection decreased the germinal vesicle breakdown rate and distorted MI spindle organization. Our results indicate that Plk1 is a pivotal regulator of microtubule organization during mouse oocyte meiosis, fertilization, and cleavage and that its functions may be regulated by other kinases, such as staurosporine-sensitive kinases.
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
Chao
C
Tong
Heng-Yu
HY
Fan
Li
L
Lian
Shi-Wen
SW
Li
Da-Yuan
DY
Chen
Heide
H
Schatten
Qing-Yuan
QY
Sun
D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D015153:Blotting, Western; D029721:Drosophila Proteins; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro; D018858:Germinal Center / Q000502:physiology; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D008845:Microinjections; D018613:Microscopy, Confocal; D008870:Microtubules / Q000502:physiology*; D008938:Mitosis / Q000502:physiology; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D008941:Spindle Apparatus / Q000187:drug effects / Q000502:physiology; D014404:Tubulin / Q000096:biosynthesis
D019138:Antibodies, Blocking; D029721:Drosophila Proteins; D014404:Tubulin; D017346:Protein Serine-Threonine Kinases