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Hum. Reprod. Advance Access originally published online on March 20, 2006
Human Reproduction 2006 21(7):1765-1770; doi:10.1093/humrep/del074
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Meiotic spindle configuration is differentially influenced by FSH and epidermal growth factor during in vitro maturation of mouse oocytes

G. Rossi 1 , G. Macchiarelli 2 , M.G. Palmerini 2 , R. Canipari 3 and S. Cecconi 1 , 4

1 Department of Biomedical Sciences and Technologies 2 Department of Experimental Medicine, University of L’Aquila, L’Aquila 3 Department of Histology and Medical Embryology, University of Roma, ‘La Sapienza’, Roma, Italy

4 To whom correspondence should be addressed at: Department of Biomedical Sciences and Technologies, University of L’Aquila, Via Vetoio, Loc. Coppito, 67010 Coppito, L’Aquila, Italy. E-mail: cecconi{at}univaq.it

BACKGROUND: To ascertain whether different hormonal treatment protocols could affect metaphase II (MII) spindle morphology, meiotic spindle organization was detected in prepubertal mouse oocytes matured under conditions allowing spontaneous, FSH- or epidermal growth factor (EGF)-dependent meiotic maturation. METHODS: Oocyte-cumulus complexes (OCCs) were matured either spontaneously (control; n = 270) or in the presence of hypoxanthine (Hx) plus FSH (n = 400) or EGF (n = 370). Spindles were detected by immunofluorescence analysis. In vivo ovulated (IVO) oocytes were processed similarly. RESULTS: IVO oocytes displayed spindles underlying the oolemma and with focused poles marked by spots of {gamma}-tubulin, whereas the majority (89%) of control oocytes had barrel-shaped spindles, positioned away from the oolemma, and with {gamma}-tubulin distributed along microtubules. Similar configuration/localization was found in 85% of the oocytes matured in vitro in the presence of Hx and FSH. In the presence of Hx-EGF, 35% of the oocytes showed spindles with an IVO-like configuration, although {gamma}-tubulin was homogeneously distributed throughout microtubules. Independently of spindle shape, 52% of EGF-stimulated oocytes had spindles positioned near the oolemma, in comparison to just 24% of FSH-treated and 13% of control oocytes. CONCLUSIONS: These results indicate that FSH and EGF can differently affect meiotic spindle morphology, and that EGF might be a stronger contributor than FSH to the acquisition of oocyte competence.

Key words: EGF/FSH/ in vitro maturation/oocyte meiotic spindle


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