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Hui Zhu,
Yugui Cui, Jin Xie,
Ling Chen,
Xiangxiang Chen,
Xuejiang Guo,
Yefei Zhu,
Xinghai Wang,
Jiansun Tong,
Zuomin Zhou,
Yue Jia,
Yan-He Lue,
Amiya Sinha Hikim,
Christina Wang,
Ronald S Swerdloff,
Jiahao Sha
PROTEOMICS - CLINICAL APPLICATIONS 04/2011; 5(3-4):191. · 1.81 Impact Factor
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Minyue Ma,
Lin Zhou,
Xuejiang Guo,
Zhuo Lv,
Yang Yu,
Chenhui Ding,
Ping Zhang,
Ye Bi, Jin Xie,
Liu Wang,
Min Lin,
Zuomin Zhou,
Ran Huo,
Jiahao Sha,
Qi Zhou
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ABSTRACT: Compaction, occurring at the eight-cell stage of mouse development, is the process of cell flattening and polarization by which cellular asymmetry is first established. During this process many molecules and organelles undergo polarized distribution, but the cytoskeletal basis for these distribution specifications remains to be explored. The present study focused on cofilin1, an actin-binding protein that depolymerizes actin filaments. We showed that cofilin1 expression decreased at the compaction stage, and that down-regulation of cofilin1 expression by siRNA microinjection accelerated compaction. Continuous observation using time-lapse video miscroscopy confirmed these findings. That is, the embryonic cells microinjected with anti-cofilin1 antibody exhibit earlier adherence properties compared to uninjected cells. Pronuclear microinjection of a site-directed mutated cofilin1 plasmid, in which cofilin1 is sustained in its active form produced embryos with blastomeres that did not adhere, suggesting that inactivation of cofilin1 is critical for cell flattening and adherence. Fluorescein-phalloidin staining indicated that decreased cofilin1 expression promoted the formation of the apical pole, which is a marker for polarity. Scanning electron microscopy results demonstrated the appearance of microvilli on the outer face of blastomeres in cofilin1 knockdown embryos. Our results suggest that cofilin1 plays an important role in cortical cytoplasmic organization during embryo compaction.
Biochimica et Biophysica Acta 09/2009; 1793(12):1804-10. · 4.66 Impact Factor
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Ningling Wang,
Ping Zhang,
Xuejiang Guo, Jin Xie,
Ran Huo,
Fuqiang Wang,
Lin Chen,
Jian Shen,
Zuomin Zhou,
Qinghua Shi,
Baige Zhao,
Jiahao Sha
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ABSTRACT: The assembly of primordial follicles early in ovarian development and subsequent transition to primary follicles are critical processes in ovarian biology. Inappropriate coordination of these processes contributes to ovarian pathologies such as premature ovarian failure and infertility. To better understand the molecular mechanisms involved in primordial follicle assembly and development, 2-D PAGE and MALDI-TOF/TOF technologies were used to construct a comparative proteome profile of the immature rat ovary at specific time-points (0, 24, 48, and 72 h postpartum). A total of 154 differential protein spots corresponding to 134 different proteins were definitively identified between any two time-points. Further cluster analysis showed four expression patterns, and each pattern correlated with specific cell processes that occur during early ovarian development. Seven proteins were randomly selected to verify expression patterns using Western blotting, and subsequently immunohistochemistry was performed to further investigate their cellular localization. Additionally, detailed functional analyses of these differentially expressed proteins were performed. Elucidation of how these changes in protein expression level coordinate primordial follicles assembly and development is intended to provide a better understanding of these critical biological processes early in ovarian development and will provide potential therapeutic molecular targets to regulate ovarian function and treat ovarian disease.
Proteomics 06/2009; 9(13):3425 - 3434. · 4.43 Impact Factor
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Xiao-Yan Huang,
Xue-Jiang Guo,
Jian Shen,
Yu-Feng Wang,
Lin Chen, Jin Xie,
Ning-Ling Wang,
Fu-Qiang Wang,
Chun Zhao,
Ran Huo,
Min Lin,
Xinru Wang,
Zuo-Min Zhou,
Jia-Hao Sha
[show abstract]
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ABSTRACT: Spermatogenesis is a complex process of terminal differentiation wherein mature sperm are produced. In the first wave of mouse spermatogenesis, different spermatogenic cells appear at specific time points, and their appearance is expected to be accompanied by changes in specific protein expression patterns. In this study, we used 2D-PAGE and MALDI-TOF/TOF technology to construct a comparative proteome profile for mouse testis at specific time points (days 0, 7, 14, 21, 28, and 60 postpartum). We identified 362 differential protein spots corresponding to 257 different proteins. Further cluster analysis revealed 6 expression patterns, and bioinformatics analysis revealed that each pattern was related to many specific cell processes. Among them, 28 novel proteins with unknown functions neither in somatic cells nor germ cells were identified, 8 of which were found to be uniquely or highly expressed in mouse testes via comparison with the GNF SymAtlas database. Further, we randomly selected 7 protein spots and the above 8 novel proteins to verify the expression pattern via Western blotting and RT-PCR, and 6 proteins with little information in testis were further investigated to explore their cellular localization during spermatogenesis by performing immunohistochemistry for the mouse testis tissue. Taken together, the above results reveal an important proteome profile that is functional during the first wave of mouse spermatogenesis, and they provide a strong basis for further research.
Journal of Proteome Research 07/2008; 7(8):3435-46. · 5.11 Impact Factor
