Shao-Chen Sun, Ph. D. , Wei-Wei Gao, M. S. , Yong-Nan Xu, Ph. D

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Presentation transcript:

Degradation of actin nucleators affects cortical polarity of aged mouse oocytes Shao-Chen Sun, Ph.D., Wei-Wei Gao, M.S., Yong-Nan Xu, Ph.D., Yong-Xun Jin, Ph.D., Qing-Ling Wang, Ph.D., Xi-Jun Yin, Ph.D., Xiang-Shun Cui, Ph.D., Nam-Hyung Kim, Ph.D. Fertility and Sterility Volume 97, Issue 4, Pages 984-990 (April 2012) DOI: 10.1016/j.fertnstert.2012.01.101 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 In vitro aging of MII oocytes for 30 hours after hCG injection causes loss of cortical polarity. (A) Expression of actin and CGs distribution and spindle morphology in fresh and aged MII oocytes. The actin cap and CG-free domain were formed in fresh MII oocytes with a normal spindle morphology. In aged MII oocytes the actin cap was weaker, and CGs were distributed throughout the entire cortex, with an abnormal morphology of the spindle. Bar = 20 μm. (B) Three-dimensional analysis of the fluorescence intensity of actin and CGs in fresh and aged MII oocytes. Fertility and Sterility 2012 97, 984-990DOI: (10.1016/j.fertnstert.2012.01.101) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Expression and localization of actin nucleators in aged oocytes. (A) Subcellular localization of ARP2 in MII oocytes as revealed by ARP antibody staining. In aged oocytes the expression of ARP2 was significantly weaker in the cytoplasm and the cortex. Green, ARP2; red, actin; blue, chromatin. (B) Fluorescence intensity of ARP2 in aged MII oocytes. (C) Abnormal rate of ARP2 expression in aged oocytes. (D) Subcellular localization of JMY in MII oocytes as revealed by JMY antibody staining. In aged oocytes the spindle localization of JMY was also significantly weaker. Green, JMY; red, actin; blue, chromatin. (E) Fluorescence intensity of JMY in aged MII oocytes. (F) Abnormal rate of JMY expression in aged oocytes. (G) Subcellular localization of WAVE2 in MII oocytes as revealed by WAVE2 antibody staining. In aged oocytes no enrichment of WAVE2 was observed near the cortex. Green, WAVE2; blue, chromatin. (H) Fluorescence intensity of WAVE2 in aged MII oocytes. (I) Rate of abnormal WAVE2 expression in aged oocytes. Fertility and Sterility 2012 97, 984-990DOI: (10.1016/j.fertnstert.2012.01.101) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Expression of ARP2, JMY, and WAVE2 in caffeine-rescue aged oocytes. Each sample contained 30 oocytes treated with 10 mM caffeine to study the expression of ARP2, JMY, and WAVE2. The expression of actin nucleation factors was normal in the oocytes treated with caffeine. Fertility and Sterility 2012 97, 984-990DOI: (10.1016/j.fertnstert.2012.01.101) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Levels of Arpc2, Arpc3, Jmy, and Wave2 mRNA in aged oocytes. mRNA expression was determined by real-time RT-PCR analysis. Each sample contained 30 oocytes. Arpc2, Arpc3, Jmy, and Wave2 mRNA expression was significantly lower in aged oocytes than in fresh oocytes. Fertility and Sterility 2012 97, 984-990DOI: (10.1016/j.fertnstert.2012.01.101) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions