Morphological and molecular markers are correlated with maturation-competence of human oocytes

STUDY QUESTION Does the position of the germinal vesicle (GV) in human oocytes correlate with molecular and morphological parameters as well as with maturation-competence? SUMMARY ANSWER The position of GV in human oocytes correlates with density of microtubule (MT) filaments, concentration of Fyn,...

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Veröffentlicht in:Human reproduction (Oxford) 2013-09, Vol.28 (9), p.2482-2489
Hauptverfasser: Levi, M., Ghetler, Y., Shulman, A., Shalgi, R.
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Sprache:eng
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Zusammenfassung:STUDY QUESTION Does the position of the germinal vesicle (GV) in human oocytes correlate with molecular and morphological parameters as well as with maturation-competence? SUMMARY ANSWER The position of GV in human oocytes correlates with density of microtubule (MT) filaments, concentration of Fyn, nucleolus localization and the ability of the oocytes to complete maturation following GV breakdown (GVBD). WHAT IS KNOWN ALREADY Our knowledge is confined to oocytes of young mice where maturation-competence is correlated with a central GV and regulated by MTs and the presence of a chromatin ring. Fyn kinase is localized at the spindle and cortex of mouse oocytes and plays a role in both maturation and MT stabilization. STUDY DESIGN, SIZE, DURATION Spatial localization of the GV and nucleolus (central or peripheral), the presence of a chromatin ring, the localization of Fyn, MT density and oocyte maturation were assessed in 153 human oocytes, 335 oocytes from young mice (2-month-old) and 146 oocytes from old mice (12-month-old). PARTICIPANTS/MATERIALS, SETTING, METHODS GV human oocytes were donated by consenting female patients (n = 57), 21–45-year-old undergoing IVF/ICSI. As a control, GV mouse oocytes were collected from female mice after injection of pregnant mares' serum gonadotrophin. Human and mouse GV oocytes allocated for immunocytochemistry were fixed on day of retrieval, stained with specific antibodies and imaged using a confocal laser-scanning microscope. Human and mouse oocytes allocated for maturation were incubated for 48 and 24 h, respectively. GVBD and extrusion of the first polar body (PBI) were assessed using differential interference contrast optics. MAIN RESULTS AND THE ROLE OF CHANCE GV location was peripheral and independent of age in 69.9% of the human oocytes, but GV location did vary with age in mice oocytes; it was central in 89.9% of the oocytes retrieved from young-mice and peripheral in 52.1% of the oocytes retrieved from old mice (P < 0.05). A central GV, whether in human or mouse oocytes, was highly correlated with a central nucleolus, absence of Fyn at the GV and a dense MT network (P < 0.05), whereas a peripheral GV correlated with peripheral nucleolus, presence of Fyn at the GV and a flimsy MT network. After 48 h in culture, no degeneration was observed in human central-GV oocytes, however, 12/95 (12.6%) of the peripheral-GV oocytes degenerated (P < 0.05). No correlation was observed between GV position and presence of a chrom
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/det261