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Subcellular localisation of key enzymes that regulate mammalian egg maturationEdgecumbe, Paul Christopher Unknown Date (has links)
Meiosis requires cells to undergo two successive rounds of division without an intermediate synthesis or S-phase. This results in a gamete with half the number of chromosomes that can then combine with those of another gamete to begin the development of a new offspring. Once an oocyte enters meiosis it is arrested in late prophase I and remains in that state until the animal reaches sexual maturity and undergoes ovulation. Upon hormonal stimuli, the oocyte is released from its first meiotic arrest, undergoes oocyte maturation, arrests in metaphase II and is ovulated to await fertilisation. The process of oocyte maturation is governed by the interaction of many different protein kinases, phosphatases and proteases. How these proteins co-localise with each other during oocyte maturation is a central aim of this thesis. One central protein is maturation promoting factor or MPF. MPF is composed of a catalytic subunit, cdc2 and a regulatory subunit, cyclin B1. Throughout oocyte maturation MPF activity is dynamic and changes in MPF activity coordinate the progression of oocyte maturation. The ubiquitin-proteasome pathway (UPP) plays a role in regulating the activity of MPF at specific times during oocyte maturation by targeting and degrading the cyclin B1 subunit. In this thesis the 20S proteasome ‘core’ of the 26S proteasome was localised in bovine and murine oocytes undergoing maturation in vitro. The 20S proteasome was found in the germinal vesicle prior to maturation in both species. Once germinal vesicle breakdown occurred, the proteasome consistently localised around the developing spindle in both species. Cyclin B1, an important target of the UPP was also localised in murine oocytes to see how the UPP interacted with its substrates during oocyte maturation. Cyclin B1 was found to have similar localisation patterns throughout oocyte maturation to the 20S proteasome. This demonstrates that the 26S proteasome moves to the location of its substrates in order to degrade them. To confirm the role of the UPP in cyclin B1 degradation, we inhibited proteasome function using MG132, a known inhibitor of proteasome function. Polar body 1 (PB1) extrusion declined significantly (P<0.05) in a dose dependent manner, confirming that the UPP is essential for the continuation of meiosis. Our localisation data indicated that cyclin B1 interacted with the UPP during the GV stage and around GVBD. Inhibition of the proteasome, again using MG132, prevented oocytes from undergoing GVBD and arrested them in the GV stage. This demonstrated that the UPP was essential for oocyte maturation as well as indicating a potential switch in substrate during this transition. The deubiquitinylating enzyme, Fat Facets in Mouse (FAM also know as USP9x), an antagonist of the UPP, was also localised in murine oocytes undergoing maturation in vitro. FAM was found primarily in the cytoplasm prior to germinal vesicle breakdown however it relocalised to the developing spindle upon germinal vesicle breakdown. FAM is known to rescue a number of specific substrates from ubiquitinylation and degradation, one being I- catenin, a multifunctional protein, not previously implicated in oocyte maturation. Localisation of I-catenin during oocyte maturation revealed distinct staining at specific stages of meiosis. Staining was observed on the plasma membrane and in the GV prior to maturation. During MI and MII it appeared to co-localise with FAM. When homologous chromosomes divided at AI/TI I-catenin distinctly localised to the cleavage furrow indicating its involvement during cytokinesis. To determine if FAM activity was essential for oocyte maturation we inhibited FAM function by microinjecting anti-FAM serum into eggs post GVBD. Inhibition of FAM prevented PB1 extrusion. Some oocytes appeared to attempt PB1 but failed to complete (Hoechst staining of the chromatin revealed these cells remained in MI). This finding strongly indicates that FAM maybe rescuing I-catenin from degradation during MI until it is needed at AI/TI. However further examination of this interaction are needed to clarify the data. This research demonstrates that regulation of turnover of key proteins is essential for progression through oocyte maturation. It also demonstrates that multiple pathways may regulate the same processes. This may add to the complexity of studying the cell, but it also demonstrates the elegance of a highly regulated and sophisticated system.
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The effects of some peptide hormones on osteoblast-like cells : with specific focus on oxytocin and vasopressin /Lagumdzija, Alena, January 2005 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.
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Biochemical analysis of cell division protein complexes in Streptomyces coelicolorKotun, Allen M. January 2007 (has links) (PDF)
Thesis (M.S.)--Duquesne University, 2007. / Title from document title page. Abstract included in electronic submission form. Includes bibliographical references (p. 81-88) and appendix.
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Developmental control of cell division in Streptomyces coelicolor /Grantcharova, Nina, January 2006 (has links)
Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 4 uppsatser.
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Investigating the role of gamma-tubulin in coordinating microtubule plus end behaviour with regulation at the spindle poleCuschieri, Lara Marie. January 1900 (has links)
Thesis (Ph.D.). / Written for the Dept. of Biology. Title from title page of PDF (viewed 2008/01/12). Includes bibliographical references.
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CEACAM1 : a common regulator of fat metabolism and cell proliferationLee, Sang Jun. January 2008 (has links)
Dissertation (Ph.D.)--University of Toledo, 2008. / "In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Sciences." Title from title page of PDF document. Bibliography: p. 74-82, 116-124, 146-192.
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Role of the mitotic spindle in the equal segregation of an extrachromosomal element in Saccharomyces cerevisiaeCui, Hong, January 1900 (has links)
Thesis (Ph. D.)--University of Texas at Austin, 2008. / Vita. Includes bibliographical references.
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The effect of sulfhydryl compounds on the sensitivity of interphase and dividing stages of Tetrahymena pyriformis W. to ultraviolet lightSullivan, William Daniel. January 1959 (has links)
Thesis--Catholic University of America. / Reprinted from Transactions of the American Microscopical Society, v. 78, no. 2-3, 1959.
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Unraveling the tangled skein the functions of redundant transcriptional regulators in cell division, intestinal homeostasis, and stress /Kirienko, Natalia V. January 2009 (has links)
Thesis (Ph.D.)--University of Wyoming, 2009. / Title from PDF title page (viewed on Apr. 16, 2010). Includes bibliographical references (p. 200-225).
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Mitotic microtubule depolymerization and XMAP215 /Shirasu-Hiza, Michele, January 2004 (has links)
Thesis (Ph.D.)--University of California, San Francisco, 2004. / Includes bibliographical references. Also available online.
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