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• W2057140147 abstract "Oocytes are complex cells comprising many cellular systems, each of which is essential for proper oocyte function. Methods to assess the diverse and critical cellular systems in oocytes derived from in vitro maturation are badly needed. Oocytes are complex cells comprising many cellular systems, each of which is essential for proper oocyte function. Methods to assess the diverse and critical cellular systems in oocytes derived from in vitro maturation are badly needed. The article by Li et al. (1Li Y. Feng H.-L. Cao Y.-J. Zheng G.-J. Yang Y. Mullen S. et al.Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro.Fertil Steril. 2006; 85: 827-832Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar) presented in this issue of Fertility and Sterility reports on the development of immature oocytes extracted from their native environment and cultured in vitro. The investigators’ findings, that oocytes derived from in vitro maturation (IVM) exhibit more spindle malformations and chromosome misalignments than in vivo–matured oocytes highlight the delicate and labile nature of cellular systems within oocytes and the influence of the follicular environment on the oocyte’s health. During development, oocytes and cumulus cells communicate through trans-zona projections and gap junctions (2Albertini D.F. Sanfins A. Combelles C.M. Origins and manifestations of oocyte maturation competencies.Reprod Biomed Online. 2003; 6: 410-415Abstract Full Text PDF PubMed Scopus (120) Google Scholar), so the developing cumulus–oocyte complex functions as a single unit, until FSH interrupts these communications (3Combelles C.M. Carabatsos M.J. Kumar T.R. Matzuk M.M. Albertini D.F. Hormonal control of somatic cell oocyte interactions during ovarian follicle development.Mol Reprod Dev. 2004; 69: 347-355Crossref PubMed Scopus (84) Google Scholar). In vivo, the intrafollicular environment provides context for the oocyte’s development, and this relationship in turn is regulated by the hormonal milieu (3Combelles C.M. Carabatsos M.J. Kumar T.R. Matzuk M.M. Albertini D.F. Hormonal control of somatic cell oocyte interactions during ovarian follicle development.Mol Reprod Dev. 2004; 69: 347-355Crossref PubMed Scopus (84) Google Scholar). Recapitulating these conditions in vitro presents significant challenges, but advances in culture media that respect the physiology of the cumulus cells and their interactions with oocytes have begun to improve the efficiency of IVM. Cellular systems within oocytes are dynamic and diverse. For example, progression of the nucleus from the germinal vesicle stage through meiosis I and its arrest at metaphase of meiosis II occurs with precise timing (4Shimada M. Terada T. Roles of cAMP in regulation of both MAP kinase and p34(cdc2) kinase activity during meiotic progression, especially beyond the MI stage.Mol Reprod Dev. 2002; 62: 124-131Crossref PubMed Scopus (63) Google Scholar). Disruption in nuclear dynamics presumably underlies the majority of nondisjunction observed in preimplantation embryos and resulting spontaneous abortions (5Hassold T. Hunt P. To err (meiotically) is human the genesis of human aneuploidy.Nat Rev Genet. 2001; 2: 280-291Crossref PubMed Scopus (1715) Google Scholar). Monitoring the dynamics of the nucleus and other cellular systems within oocytes might facilitate improvement of media and the IVM process. Li et al. (1Li Y. Feng H.-L. Cao Y.-J. Zheng G.-J. Yang Y. Mullen S. et al.Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro.Fertil Steril. 2006; 85: 827-832Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar) studied the effects of IVM on the meiotic spindle and on chromosomes. They used immunofluorescence to assess spindle integrity and labeling with propridium iodide to image chromosomes. Their finding that IVM is associated with abnormal spindle integrity and chromosome congression is sobering. Like any important scientific finding, it raises more questions than it answers. Did IVM itself disrupt the spindles and chromosomes, or did it promote the survival of oocytes with abnormal spindles and chromosomes, which would not have developed outside the IVM system? Is the IVM process associated with abnormalities in other key organelles (e.g., Golgi [6Cobbold C. Coventry J. Ponnambalam S. Monaco A.P. Actin and microtubule regulation of trans-Golgi network architecture, and copper-dependent protein transport to the cell surface.Mol Membr Biol. 2004; 21: 59-66Crossref PubMed Scopus (20) Google Scholar]) or molecules (e.g., RNA [7Kloc M. Wilk K. Vargas D. Shirato Y. Bilinski S. Etkin L.D. Potential structural role of non-coding and coding RNAs in the organization of the cytoskeleton at the vegetal cortex of Xenopus oocytes.Development. 2005; 132: 3445-3457Crossref PubMed Scopus (124) Google Scholar])? Tools to assess those cellular systems noninvasively in living oocytes (1Li Y. Feng H.-L. Cao Y.-J. Zheng G.-J. Yang Y. Mullen S. et al.Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro.Fertil Steril. 2006; 85: 827-832Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar) would help answer these questions. Oocytes are complex cells comprising many organelles and molecules, each of which must be in the appropriate “state” for the egg to be competent to sustain subsequent embryo development. Just as the car’s battery must be charged, its tires inflated, and its transmission engaged, similarly the organelles and molecules within the oocyte must be primed for development. The normal egg exhibits [1] chromosomes aligned at the metaphase plate and microtubules arranged into a barrel-shaped anastral spindle (8Keefe D. Liu L. Wang W. Silva C. Imaging meiotic spindles by polarization light microscopy principles and applications to IVF.Reprod Biomed Online. 2003; 7: 24-29Abstract Full Text PDF PubMed Scopus (95) Google Scholar), [2] mitochondria with condensed cristae localized peripherally (9Au H.K. Yeh T.S. Kao S.H. Tzeng C.R. Hsieh R.H. Abnormal mitochondrial structure in human unfertilized oocytes and arrested embryos.Ann N Y Acad Sci. 2005; 1042: 177-185Crossref PubMed Scopus (54) Google Scholar), [3] cortical granules docked at the plasma membrane (10Sun Q.Y. Cellular and molecular mechanisms leading to cortical reaction and polyspermy block in mammalian eggs.Microsc Res Tech. 2003; 61: 342-348Crossref PubMed Scopus (85) Google Scholar), [4] Golgi apparatus distributed in a punctate pattern throughout the cytoplasm (11Moreno R.D. Schatten G. Ramalho-Santos J. Golgi apparatus dynamics during mouse oocyte in vitro maturation effect of the membrane trafficking inhibitor brefeldin A.Biol Reprod. 2002; 66: 1259-1266Crossref PubMed Scopus (38) Google Scholar), and [5] messenger RNA stored in appropriate quantity and location (12Bermudez M.G. Wells D. Malter H. Munne S. Cohen J. Steuerwald N.M. Expression profiles of individual human oocytes using microarray technology.Reprod Biomed Online. 2004; 8: 325-337Abstract Full Text PDF PubMed Scopus (68) Google Scholar). Which of these factors determines the developmental capacity of the oocyte? Just as the defective car will not drive or, more seriously, will result in a collision, so dysfunction involving the oocyte’s various components can result in failed fertilization, arrested cleavage, failed implantation, or worse, development of a handicapped offspring. As with the components of a car, not every system of an egg requires individual assessment. Rather, we need metrics to evaluate the “state” of the key cellular systems within the oocyte. For example, oxygen consumption or metabolite use by oocytes could reflect mitochondrial function (13Trimarchi J.R. Liu L. Porterfield D.M. Smith P.J. Keefe D.L. Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos.Biol Reprod. 2000; 62: 1866-1874Crossref PubMed Scopus (196) Google Scholar, 14Gardner D.K. Lane M. Stevens J. Schoolcraft W.B. Noninvasive assessment of human embryo nutrient consumption as a measure of developmental potential.Fertil Steril. 2001; 76: 1175-1180Abstract Full Text Full Text PDF PubMed Scopus (244) Google Scholar). By understanding which cellular systems drive development, we can “move the lamppost” to key areas that provide accurate assessments of oocyte quality and developmental competence. Two aspects of the study by Li et al. (1Li Y. Feng H.-L. Cao Y.-J. Zheng G.-J. Yang Y. Mullen S. et al.Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro.Fertil Steril. 2006; 85: 827-832Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar) should be more thoroughly considered before we can fully accept their findings: the methodology of IVM and the investigators’ approach to assessing spindle integrity and chromosome alignment. Li et al.’s findings that oocytes derived from IVM exhibited disrupted spindles and misaligned chromosomes might not be generalizable to all IVM techniques. Theoretically, some IVM media might better emulate the natural intrafollicular environment and therefore produce oocytes with cellular systems in the same state as those developed in vivo. Indeed, tissue engineering technology might improve the efficiency of IVM and enhance oocyte quality over that which can be obtained in vivo (15Combelles C.M. Fissore R.A. Albertini D.F. Racowsky C. In vitro maturation of human oocytes and cumulus cells using a co-culture three-dimensional collagen gel system.Hum Reprod. 2005; 20: 1349-1358Crossref PubMed Scopus (84) Google Scholar, 16Nogueira D. Cortvrindt R. Everaerdt B. Smitz J. Effects of long-term in vitro exposure to phosphodiesterase type-3 inhibitors on follicle and oocyte development.Reproduction. 2005; 130: 177-186Crossref PubMed Scopus (28) Google Scholar). The observations by Li et al. highlight the importance of understanding the effects of IVM on development, so we can improve the IVM technique. Heisenberg articulated that the act of taking a measurement from a system can impact that system, thereby limiting the accuracy of the measurement. Such is the case with the study of living cells, and in particular the study of spindle integrity. Spindles are exquisitely labile polymers of tubulin that rapidly disassemble during fixation and processing for immunostaining (8Keefe D. Liu L. Wang W. Silva C. Imaging meiotic spindles by polarization light microscopy principles and applications to IVF.Reprod Biomed Online. 2003; 7: 24-29Abstract Full Text PDF PubMed Scopus (95) Google Scholar, 17Sanfins A. Plancha C.E. Overstrom E.W. Albertini D.F. Meiotic spindle morphogenesis in in vivo and in vitro matured mouse oocytes insights into the relationship between nuclear and cytoplasmic quality.Hum Reprod. 2004; 19: 2889-2899Crossref PubMed Scopus (64) Google Scholar). It is likely that some of the abnormal spindles identified by Li et al. arose from artifacts of fixation and staining. Application of alternative methods to label and image spindle, such as fluorescent labeling of tubulin and polarized light microscopy, would help confirm their important findings. In summary, we know that IVM as a standard protocol for bovine assisted reproduction delivers excellent results, and therefore likely eventually will be perfected to the point that it will produce normal human oocytes. To expedite development of effective IVM culture approaches, we desperately need more studies, such as that published in this issue of Fertility and Sterility. Moreover, we need better methods to quantitatively assess the state of cellular systems in oocytes." @default.
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• W2057140147 title "Assessing the quality of oocytes derived from in vitro maturation: are we looking under the lamppost?" @default.
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