TY - JOUR
T1 - The involvement of Fyn kinase in resumption of the first meiotic division in mouse oocytes
AU - Levi, Mattan
AU - Maro, Bernard
AU - Shalgi, Ruth
N1 - Funding Information:
This work was supported by a grant from the Israel Science Foundation to R. Shalgi and by the Eshkol scholarship granted by the Israel Ministry of Science, Culture and Sport to M. Levi. We thank Dr. J.Z. Kubiak for critical reading of the paper. We thank Dr. M.H. Verlhac, Pierre and Marie Curie University, Paris, for the Histone-H2B-RFP and β-tubulin-GFP plasmids and for the anti-ERK1/2 antibody. We also thank R. Kaplan-Kraicer for her help in preparing the manuscript. This work is a partial fulfillment of the requirements for the Ph.D. degree of M. Levi at the Sackler Faculty of Medicine, Tel-Aviv University.
PY - 2010/4/15
Y1 - 2010/4/15
N2 - The process of resumption of the first meiotic division (RMI) in mammalian oocytes includes germinal vesicle breakdown (GVBD), spindle formation during first metaphase (MI), segregation of homologous chromosomes, extrusion of the first polar body (pBI) and an arrest at metaphase of the second meiotic division (MII). previous studies suggest a role for Fyn, a non-receptor Src family tyrosine kinase, in the exit from MII arrest. In the current study we characterized the involvement of Fyn in RMI. Western blot analysis demonstrated a significant, proteasome independent, degradation of Fyn during GVBD. Immunostaining of fixed oocytes and confocal imaging of live oocytes microinjected with Fyn complementary RNA (cRNA) demonstrated Fyn localization to the oocyte cortex and to the spindle poles. Fyn was recruited during telophase to the cortical area surrounding the midzone of the spindle and was then translocated to the contractile ring during extrusion of pBI. GVBD, exit from MI and pBI extrusion were inhibited in oocytes exposed to the chemical inhibitor SU6656 or microinjected with dominant negative Fyn cRNA. None of the microinjected oocytes showed misaligned or lagging chromosomes during chromosomes segregation and the spindle migration and anchoring were not affected. However, the extruded pBI was of large size. Altogether, a role for Fyn in regulating several key pathways during the first meiotic division in mammalian oocytes is suggested, particularly at the GV and metaphase checkpoints and in signaling the ingression of the cleavage furrow.
AB - The process of resumption of the first meiotic division (RMI) in mammalian oocytes includes germinal vesicle breakdown (GVBD), spindle formation during first metaphase (MI), segregation of homologous chromosomes, extrusion of the first polar body (pBI) and an arrest at metaphase of the second meiotic division (MII). previous studies suggest a role for Fyn, a non-receptor Src family tyrosine kinase, in the exit from MII arrest. In the current study we characterized the involvement of Fyn in RMI. Western blot analysis demonstrated a significant, proteasome independent, degradation of Fyn during GVBD. Immunostaining of fixed oocytes and confocal imaging of live oocytes microinjected with Fyn complementary RNA (cRNA) demonstrated Fyn localization to the oocyte cortex and to the spindle poles. Fyn was recruited during telophase to the cortical area surrounding the midzone of the spindle and was then translocated to the contractile ring during extrusion of pBI. GVBD, exit from MI and pBI extrusion were inhibited in oocytes exposed to the chemical inhibitor SU6656 or microinjected with dominant negative Fyn cRNA. None of the microinjected oocytes showed misaligned or lagging chromosomes during chromosomes segregation and the spindle migration and anchoring were not affected. However, the extruded pBI was of large size. Altogether, a role for Fyn in regulating several key pathways during the first meiotic division in mammalian oocytes is suggested, particularly at the GV and metaphase checkpoints and in signaling the ingression of the cleavage furrow.
KW - Exit from metaphase
KW - Fyn
KW - GVBD
KW - Meiosis
KW - Oocyte
KW - Polar body
UR - http://www.scopus.com/inward/record.url?scp=77953547292&partnerID=8YFLogxK
U2 - 10.4161/cc.9.8.11299
DO - 10.4161/cc.9.8.11299
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C2 - 20372074
AN - SCOPUS:77953547292
SN - 1538-4101
VL - 9
SP - 1577
EP - 1589
JO - Cell Cycle
JF - Cell Cycle
IS - 8
ER -