Zhang John, Liu Hui
Reproductive Endocrinology and Infertility, New Hope Fertility Center, New York, NY, USA.
Reproductive Endocrinology and Infertility, New Hope Fertility Center, New York, NY, USA.
Reprod Biomed Online. 2015 Jul;31(1):71-8. doi: 10.1016/j.rbmo.2015.03.012. Epub 2015 Apr 16.
Both the cytoplasmic and nuclear compartments are essential for the acquisition of meiotic competence. This study assessed the role of the cytoplasm in meiosis resumption in meiotically arrested oocytes at the germinal vesicle (GV) stage. Mouse oocytes at GV stage were meiotically arrested with 3-isobutyl-1-methylxanthine (IBMX). GV transfer was performed between IBMX-treated and non-treated (control) mouse oocytes, and between control mouse and human GV oocytes. Extrusion of first polar body (PB) was examined as an indication of nuclear maturation. Meiotic spindle assembly and chromosome alignment were examined by immunostaining. Results indicated that oocytes arrested with IBMX for 24 and 48 h exhibited reduced ability for meiotic maturation and for extruding the first PB when compared with controls (P < 0.01). IBMX-treated oocytes reconstituted with cytoplasm, but not GV, of control oocytes restored the assembly of meiotic spindle and meiotic maturation. Mouse oocytes reconstituted with GV of human oocytes underwent meiosis similar to that observed in mice, but not humans. Additionally, human oocytes reconstituted by mouse GV underwent meiosis similar to that observed in humans, but not mice. These findings suggest that cytoplasm replacement by GV transfer could represent a potential therapeutic option for women who do not produce mature oocytes during IVF.
细胞质和细胞核区室对于获得减数分裂能力均至关重要。本研究评估了细胞质在生发泡(GV)期减数分裂阻滞的卵母细胞减数分裂恢复中的作用。用3 - 异丁基 - 1 - 甲基黄嘌呤(IBMX)使GV期小鼠卵母细胞发生减数分裂阻滞。在经IBMX处理的和未处理的(对照)小鼠卵母细胞之间,以及对照小鼠和人GV卵母细胞之间进行GV转移。检查第一极体(PB)的排出作为核成熟的指标。通过免疫染色检查减数分裂纺锤体组装和染色体排列。结果表明,与对照相比,用IBMX阻滞24小时和48小时的卵母细胞减数分裂成熟和排出第一PB的能力降低(P < 0.01)。用对照卵母细胞的细胞质而非GV重建的经IBMX处理的卵母细胞恢复了减数分裂纺锤体组装和减数分裂成熟。用人GV重建的小鼠卵母细胞经历的减数分裂与在小鼠中观察到的相似,但与在人中观察到的不同。此外,用小鼠GV重建的人卵母细胞经历的减数分裂与在人中观察到的相似,但与在小鼠中观察到的不同。这些发现表明,通过GV转移进行细胞质替代可能是体外受精期间未产生成熟卵母细胞的女性的一种潜在治疗选择。