Hum. Reprod. Advance Access published online on September 23, 2004
Human Reproduction, doi:10.1093/humrep/deh528
© 2004 by European Society of Human Reproduction and Embryology
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1 Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, MA 02111, USA; Instituto de Medicina Molecular, Unidade de Biologia da Reprodução, Faculdade de Medicina de Lisboa, 1649-028 Lisboa, Portugal
* To whom correspondence should be addressed. E-mail: dalbertini{at}kumc.edu.
BACKGROUND: This work addresses the hypothesis that events occurring within the follicle soon after the LH surge are essential for coordinating morphogenesis of the spindle and cytoplasm in mouse oocytes matured in vivo (IVO); we further tested whether in vitro maturation (IVM) fails to support these events. METHODS: Oocytes collected at 1, 2, 3, 4 and 5 h post-hCG or after IVM were analyzed for chromatin, nuclear lamina, microtubules (MTs) and centrosomal proteins by conventional fluorescence and confocal microscopy. In addition, these parameters were monitored in oocytes maintained in 50 µM roscovitine, followed by IVM, or in oocytes retrieved at 1.5 and 5 h post-hCG in vivo and cultured up to 16 h. RESULTS: A G2/M delay was observed in IVO oocytes based upon persistence of cytoplasmic MTs, nuclear lamina and centrosomes at the cortex; rapid meiotic progression in IVM oocytes was related to loss of these markers, indicating that a global activation of MPF occurred in culture. Also, maturating-promoting factor (MPF) inactivation resulted in cultured oocytes that exhibited IVO characteristics after drug removal. IVO-like characteristics were also exhibited by oocytes retrieved at 5 but not at 1.5 h after hCG treatment, even though these oocytes were subsequently cultured. CONCLUSIONS: The results emphasize the importance of coupling MT remodeling and cell cycle components during oocyte maturation to achieve a balanced coordination of nuclear and cytoplasmic maturation that under physiological conditions occurs within the first 5 h of LH stimulation. 4Present address: Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160-7401, USA 5Present address: Department of Biology and Biotechnology, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA 01609-2280, USA
Accepted August 26, 2004
Article
Meiotic spindle morphogenesis in in vivo and in vitro matured mouse oocytes: insights into the relationship between nuclear and cytoplasmic quality
2 Instituto de Medicina Molecular, Unidade de Biologia da Reprodução, Faculdade de Medicina de Lisboa, 1649-028 Lisboa, Portugal
3 Department of Biomedical Sciences, Tufts University School of Veterinary Medicine, Grafton, 01536 MA, USA
4 Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, MA 02111, USA
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