Proline-rich tyrosine kinase2 is involved in F-actin organization during in vitro maturation of rat oocyte.
Microfilaments (actin filaments) regulate various dynamic events during meiotic maturation. Relatively, little is known about the regulation of microfilament organization in mammalian oocytes. Proline-rich tyrosine kinase2 (Pyk2), a protein tyrosine kinase related to focal adhesion kinase (FAK) is essential in actin filaments organization. The present study was to examine the expression and localization of Pyk2, and in particular, its function during rat oocyte maturation. For the first time, by using Western blot and confocal laser scanning microscopy, we detected the expression of Pyk2 in rat oocytes and found that Pyk2 and Try402 phospho-Pyk2 were localized uniformly at the cell cortex and surrounded the germinal vesicle (GV) or the condensed chromosomes at the GV stage or after GV breakdown. At the metaphase and the beginning of anaphase, Pyk2 distributed asymmetrically both in the ooplasm and the cortex with a marked staining associated with the chromosomes and the region overlying the meiotic spindle. At telophase, Pyk2 was observed in the cleavage furrows in addition to its cortex and cytoplasm localization. The dynamics of Pyk2 were similar to that of F-actin, and this kinase was found to co-localize with microfilaments in several developmental stages during rat oocyte maturation. Microinjection of Pyk2 antibody demolished the microfilaments assembly and also inhibited the first polar body (PB1) emission. These findings suggest an important role of Pyk2 for rat oocyte maturation by regulating the organization of actin filaments.
Meng
Xiao-Qian
XQ
Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, 88 East Wenhua Road, Jinan 250002, People's Republic of China.
Zheng
Ke-Gang
KG
Yang
Yong
Y
Jiang
Man-Xi
MX
Zhang
Yan-Ling
YL
Sun
Qing-Yuan
QY
Li
Yun-Long
YL
D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000199:Actins / Q000032:analysis / Q000648:ultrastructure*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D051418:Focal Adhesion Kinase 2 / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008845:Microinjections; D018613:Microscopy, Confocal / Q000379:methods; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D009866:Oogenesis / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar
D000199:Actins; D000911:Antibodies, Monoclonal; D051418:Focal Adhesion Kinase 2