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ROLE OF 14-3-3 ETA AND EPSILON IN GAMETOGENESIS

Eisa, Alaa Abdulaziz
http://orcid.org/0000-0002-8973-5948
http://rave.ohiolink.edu/etdc/view?acc_num=kent1574096506246273

Abstract Details

2019, PHD, Kent State University, College of Arts and Sciences / School of Biomedical Sciences.
Gametogenesis involves the interplay of a number of signaling processes and regulatory proteins. YWHA or 14-3-3 proteins are key regulatory proteins found in eukaryotic cells that, among other functions, are central in regulating the cell cycle. The 14-3-3 proteins form homodimers or heterodimers and act as adaptor proteins that affect protein localization, confirmation, stability, or activity. The family of 14-3-3 proteins, encoded by seven different genes, includes YWHAB, YWHAE, YWHAG, YWHAH, YWHAQ, YWHAZ, and SFN). Transgenic mice containing LoxP sites to remove exon 2 from the 14-3-3 eta (YWHAH) and exons 3 and 4 from 14-3-3 epsilon (YWHAE) mice were used in this study to define roles for these two isoforms in germ cells during spermatogenesis and oogenesis. The transgenic mice were bred with ACTB Cre mice for global knockout, Stra8 Cre mice for the male germ cell-specific knockout, or Zp3 Cre mice for the oocyte-specific knockout. Confirmation of the transgenic genotypes was accomplished through PCR amplification using specific primers targeted to the coding region. The absence of the protein in sperm and oocytes was confirmed by using both western blot and immunohistochemical staining techniques. In vivo breeding tests and in vitro fertilization indicate that, in the absence of 14-3-3 epsilon, males are infertile. However, mice lacking 14-3-3 eta were normal and fertile. Females lacking either of the two 14-3-3 isoforms or both does not appear to alter oogenesis, oocyte maturation or fertility. Low sperm count with higher abnormal sperm was seen in 14-3-3 epsilon knockout mice. Using the Computer Assisted Semen Analysis (CASA) system, the motility of 14-3-3 epsilon knockout sperm was seen to be significantly lower compared to the control sperm. A decrease in the phosphorylation of both glycogen synthase kinase 3 (GSK3) and Protein Phosphatase1γ2 (PP1γ2), the signal enzymes essential for male fertility, were seen in sperm from 14-3-3 epsilon knockout mice, suggesting that the absence of 14-3-3 epsilon may alter signaling pathways known to regulate spermatogenesis, sperm motility, and fertility. 14-3-3 proteins often act interchangeable in interacting with target proteins or in forming heterodimers. Here we find that 14-3-3 epsilon alone may be required for normal sperm function. We suspect that other 14-3-3 isoforms function critically in oogenesis.
Douglas Kline, PhD (Advisor)
Srinivasan Vijayaraghavan, PhD (Advisor)
Gail Fraizer, PhD (Committee Member)
Kristy Welshhans, PhD (Committee Member)
Soumitra Basu, PhD (Other)
192 p.
Biology; Biomedical Research; Developmental Biology; Molecular Biology; Physiology
14-3-3, YWHA, Spermatogenesis, Spermiogenesis, Sperm motility, Oogenesis, Oocyte maturation

Recommended Citations

Citations

  • Eisa, A. A. (2019). ROLE OF 14-3-3 ETA AND EPSILON IN GAMETOGENESIS [Doctoral dissertation, Kent State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=kent1574096506246273

    APA Style (7th edition)

  • Eisa, Alaa. ROLE OF 14-3-3 ETA AND EPSILON IN GAMETOGENESIS. 2019. Kent State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=kent1574096506246273.

    MLA Style (8th edition)

  • Eisa, Alaa. "ROLE OF 14-3-3 ETA AND EPSILON IN GAMETOGENESIS." Doctoral dissertation, Kent State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=kent1574096506246273

    Chicago Manual of Style (17th edition)

kent1574096506246273
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This open access ETD is published by Kent State University and OhioLINK.