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Meiotic Spindle (meiotic + spindle)
Selected AbstractsArabidopsis thaliana protein, ATK1, is a minus-end directed kinesin that exhibits non-processive movementCYTOSKELETON, Issue 3 2002Adam I. Marcus Abstract The microtubule cytoskeleton forms the scaffolding of the meiotic spindle. Kinesins, which bind to microtubules and generate force via ATP hydrolysis, are also thought to play a critical role in spindle assembly, maintenance, and function. The A. thaliana protein, ATK1 (formerly known as KATA), is a member of the kinesin family based on sequence similarity and is implicated in spindle assembly and/or maintenance. Thus, we want to determine if ATK1 behaves as a kinesin in vitro, and if so, determine the directionality of the motor activity and processivity character (the relationship between molecular "steps" and microtubule association). The results show that ATK1 supports microtubule movement in an ATP-dependent manner and has a minus-end directed polarity. Furthermore, ATK1 exhibits non-processive movement along the microtubule and likely requires at least four ATK1 motors bound to the microtubule to support movement. Based on these results and previous data, we conclude that ATK1 is a non-processive, minus-end directed kinesin that likely plays a role in generating forces in the spindle during meiosis. Cell Motil. Cytoskeleton 52:144,150, 2002. © 2002 Wiley-Liss, Inc. [source] Transitions in the evolution of meiosisJOURNAL OF EVOLUTIONARY BIOLOGY, Issue 3 2000Hurst Meiosis may have evolved gradually within the eukaryotes with the earliest forms having a one-step meiosis. It has been speculated that the putative transition from a one-step meiosis without recombination to one with recombination may have been stimulated by the invasion of Killer alleles. These imaginary selfish elements are considered to act prior to recombination. They prime for destruction (which occurs after cell division) the half of the cell on the opposite side of the meiotic spindle. Likewise the transition from one-step to two-step meiosis might have been stimulated by a subtly different sort of imaginary distorter allele, a SisterKiller. These are proposed to act after recombination. It has yet to be established that the presence of such distorter alleles could induce the transitions in question. To investigate these issues we have analysed the dynamics of a modifier (1) of recombination and (2) of the number of steps of meiosis, as they enter a population with one-step meiosis. For the modifier of recombination, we find that invasion conditions are very broad and that persistence of Killer and modifier is likely through most parameter space, even when the recombination rate is low. However, if we allow a Killer element to mutate into one that is self-tolerant, the modifier and the nonself-tolerant alleles are typically both lost from the population. The modifier of the number of steps can invade if the SisterKiller acts at meiosis II. However, a SisterKiller acting at meiosis I, far from promoting the modifier's spread, actually impedes it. In the former case the invasion is easiest if there is no recombination. The SisterKiller hypothesis therefore fails to provide a reasonable account of the evolution of two-step meiosis with recombination. As before, the evolution of self-tolerance on the part of the selfish element destroys the process. We conclude that the conditions under which SisterKillers promote the evolution of two-step meiosis are very much more limited than originally considered. We also conclude that there is no universal agreement between ESS and modifier analyses of the same transitions. [source] Role of AMPK throughout meiotic maturation in the mouse oocyte: Evidence for promotion of polar body formation and suppression of premature activationMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 10 2010Stephen M. Downs Abstract This study was conducted to assess the role of AMPK in regulating meiosis in mouse oocytes from the germinal vesicle stage to metaphase II. Exposure of mouse cumulus cell-enclosed oocytes (CEO) and denuded oocytes (DO) during spontaneous maturation in vitro to AMPK-activating agents resulted in augmentation of the rate and frequency of polar body formation. Inhibitors of AMPK had an opposite, inhibitory effect. In addition, the AMPK inhibitor, compound C (Cmpd C) increased the frequency of oocyte activation. The stimulatory action of the AMPK-activating agent, AICAR, and the inhibitory action of Cmpd C were diminished if exposure was delayed, indicating an early action of AMPK on polar body formation. The frequency of spontaneous and Cmpd C-induced activation in CEO was reduced as the period of hormonal priming was increased, and AMPK stimulation eliminated the activation response. Immunostaining of oocytes with antibody to active AMPK revealed an association of active kinase with chromatin, spindle poles, and midbody during maturation. Immunolocalization of the ,1 catalytic subunit of AMPK showed an association with condensed chromatin and the meiotic spindle but not in the spindle poles or midbody; ,2 stained only diffusely throughout the oocyte. These data suggest that AMPK is involved in a regulatory capacity throughout maturation and helps promote the completion of meiosis while suppressing premature activation. Mol. Reprod. Dev. 77:888,899, 2010. © 2010 Wiley-Liss, Inc. [source] Eomesodermin is expressed in mouse oocytes and pre-implantation embryosMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 4 2005Josie McConnell Abstract T-box genes are a highly conserved family of genes encoding transcription factors, which share a conserved DNA binding domain (the T-box). Appropriate temporal and spatial expression of this gene family is critical for gastrulation and organogenesis in a number of species. The T-box containing gene Eomesodermin was first identified in Xenopus, where it plays a critical role in mesoderm formation. In situ analyses in mice have described the expression patterns of the mouse ortholog of this gene mEomesodermin (mEomes) at the time of implantation and during fetal development. Additional studies involving the disruption of the mEomes gene, have demonstrated an additional role for mEomes in trophoblast formation. However, these analyses did not address the possibility that maternally encoded or pre-blastocyst zygotic transcription of mEomes may also contribute to embryonic development. We show here that mEomes mRNA is present prior to blastocyst formation, and that the protein product of mEomes is associated with nuclear DNA during oocyte development and persistently localizes within all nuclei of the preimplantation embryo until the early blastocyst stage. mEomes protein is associated with the meiotic spindle in the unfertilized egg and with the mitotic spindle at each cell division. Our results are consistent with mEomesodermin having a role in early preimplantation development and inner cell mass formation in addition to its function in the trophoblast lineage. Mol. Reprod. Dev. © 2005 Wiley-Liss, Inc. [source] Cytoskeletal Changes in Oocytes and Early Embryos During in vitro Fertilization Process in MiceANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 1 2010E. Gumus Summary The cytoskeleton plays crucial roles in the development and fertilization of germ cells and in the early embryo development. The growth, maturation and fertilization of oocytes require an active movement and a correct localization of cellular organelles. This is performed by the re-organization of microtubules and actin filaments. Therefore, the aim of the present study was to determine the changes in cytoskeleton during in vitro fertilization process using appropriate immunofluorescence techniques. While the chromatin content was found to be scattered throughout the nucleus during the oocyte maturation period, it was seen only around nucleolus following the completion of the maturation. Microtubules, during oocyte maturation, were regularly distributed throughout the ooplasm which was then localized in the subcortical region of oocytes. Similarly microfilaments were scattered throughout the ooplasm during the oocyte maturation period whereas they were seen in the subcortical region around the polar body and above the meiotic spindle throughout the late developmental stages. In conclusion, those changes occurred in microtubules and microfilaments might be closely related to the re-organization of the genetic material during the oocyte maturation and early embryo development. [source] Glutamylated tubulin: Diversity of expression and distribution of isoformsCYTOSKELETON, Issue 1 2003Marie-Louise Kann Abstract Glutamylation of , and , tubulin isotypes is a major posttranslational modification giving rise to diversified isoforms occurring mainly in neurotubules, centrioles, and axonemes. Monoglutamylated tubulin isoforms can be differentially recognized by two mAbs, B3 and GT335, which both recognize either polyglutamylated isoforms. In the present study, immunoelectron microscopy and immunofluorescence analyses were performed with these two mAbs to determine the expression and distribution of glutamylated tubulin isoforms in selected biological models whose tubulin isotypes are characterized. In mouse spermatozoa, microtubules of the flagellum contain polyglutamylated isoforms except in the tip where only monoglutamylated isoforms are detected. In spermatids, only a subset of manchette microtubules contain monoglutamylated tubulin isoforms. Cytoplasmic microtubules of Sertoli cells are monoglutamylated. Mitotic and meiotic spindles of germ cells are monoglutamylated whereas the HeLa cell mitotic spindle is polyglutamylated. Three models of axonemes are demonstrated as a function of the degree and extent of tubulin glutamylation. In lung ciliated cells, axonemes are uniformly polyglutamylated. In sea urchin sperm and Chlamydomonas, flagellar microtubules are polyglutamylated in their proximal part and monoglutamylated in their distal part. In Paramecium, cilia are bi- or monoglutamylated only at their base. In all cells, centrioles or basal bodies are polyglutamylated. These new data emphasize the importance of glutamylation in all types of microtubules and strengthen the hypothesis of its role in the regulation of the intracellular traffic and flagellar motility. Cell Motil. Cytoskeleton 55:14,25, 2003. © 2003 Wiley-Liss, Inc. [source] Alterations and reversibility in the chromatin, cytoskeleton and development of pig oocytes treated with roscovitineMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 4 2003Jyh-Cherng Ju Abstract Germinal vesicle (GV) breakdown in mammalian oocytes is regulated by the activation of maturation promoting factor (MPF). We investigated a specific cdc2 kinase inhibitor, roscovitine, to maintain pig oocytes in the GV stage. Cumulus-oocyte complexes (COCs) were aspirated from slaughterhouse ovaries and cultured for 44 hr in NCSU#23 medium containing different levels of roscovitine (0, 10, 20, 30, 40, 50 ,M in Experiment 1 and 0, 40, 60, 80, 100, 120 ,M in Experiment 2). The COCs were cultured for another 44 hr after removal of the chemical. Twenty oocytes in each group were fixed at 44 hr for immunocytochemical labeling of the cytoskeleton and the rest (,20/group) were fixed at the end of 88 hr after culture. Results showed that the inhibition of the oocyte in the GV stage was not effective when 10,50 ,M (Experiment 1) of roscovitine were used (19,34%). When oocytes were released from the inhibitor, similar proportions (70,83%) of oocytes were observed in the MII or advanced stages among treatments. However, when higher concentrations of roscovitine were used (Experiment 2), significantly greater inhibitory effect was observed at the levels of 80,120 ,M with 83,91% oocytes being blocked in the GV stage when compared to the control (9%) and the 40,60 ,M (27,43%) groups (P,<,0.05). Although 15,21% of the oocytes showed abnormal MII morphology with aberrant meiotic spindles and/or formation of cytoplasmic microtubules, a substantial number of oocytes resumed meiosis and reached MII stage at 44 hr after removal of this chemical. In Experiment 3, different concentrations of roscovitine (0, 20, 40, and 80 ,M) were tested to examine the length of intervals (0, 11, 22, 33, and 44 hr) for an effective inhibition. Results showed that the inhibitory effect was significantly more prominent at 22 hr than that at 33 and 44 hr after roscovitine treatment in all treatment groups (P,<,0.05). This study demonstrated that roscovitine-treated oocytes resumed meiosis after removal of the inhibitor. This could provide flexibility for studying porcine oocyte development and embryo cloning and may have application in other species. Mol. Reprod. Dev. 64: 482,491, 2003. © 2003 Wiley-Liss, Inc. [source] | ||||||||||