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Distinctions in Meiotic Spindle Structure and Assembly During In Vitro and In Vivo Maturation of Mouse Oocytes¹

Department of Anatomy and Cellular Biology,³ Tufts University School of Medicine, Boston, Massachusetts 02111 Unidade de Biologia da Reprodução,⁴ Faculdade de Medicina de Lisboa, Lisboa, Portugal Department of Biomedical Sciences,⁵ Tufts University School of Veterinary Medicine, Grafton, Massachusetts 01536

To better understand the differences in cytoskeletal organization between in vivo (IVO) and in vitro (IVM) matured oocytes, we analyzed remodeling of the centrosome-microtubule complex in IVO and IVM mouse oocytes. Fluorescence imaging revealed dramatic differences in meiotic spindle assembly and organization between these two populations. Metaphase spindles at both meiosis I (M-I) and meiosis II (M-II) in IVO oocytes were compact, displayed focused spindle poles with distinct -tubulin foci, and were composed of acetylated microtubules. In contrast, IVM oocytes exhibited barrel-shaped spindles with fewer acetylated microtubules and -tubulin diffusely distributed throughout the spindle proper. With respect to meiotic progression, IVO oocytes were more synchronous in the rate and extent of anaphase to telophase of M-I and first polar body emission than were IVM counterparts. Furthermore, IVO oocytes showed a twofold increase in cytoplasmic microtubule organizing centers (MTOCs), and constitutive MTOC proteins (-tubulin and pericentrin) were excluded from the first polar body. Inclusion of MTOC constitutive proteins in the polar body and diminished number of cytoplasmic MTOCs was observed in IVM oocytes. These findings were corroborated in IVO oocytes obtained from naturally ovulated and spontaneously cycling mice and highlight a fundamental distinction in the spatial and temporal regulation of microtubule dynamics between IVO and IVM oocytes

¹ This work was supported by grants from Fundaçao para a Ciencia e Tecnologia SFRH/BD/2757/2000 (to A.S.), POCTI/ESP/43628/2000 (to C.E.P.), March of Dimes Birth Defects Foundation 01-248 (to D.A.), and USDA-NRI 2001-35203-09966 (to E.O. and D.A.).

² Correspondence: Alexandra Sanfins, Department of Anatomy and Cellular Biology, Tufts University School of Medicine, 136 Harrison Ave., Boston, Massachusetts 02111. FAX: 617 636 6536; alexa_sanfins{at}yahoo.com

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