Benammar Achraf, Ziyyat Ahmed, Lefèvre Brigitte, Wolf Jean-Philippe
1 Inserm U1016, CNRS UMR8104, Université Paris Descartes, Paris, France.
2 Service d'Histologie Embryologie Biologie de la Reproduction-CECOS, Hôpital Cochin, AP-HP, Paris, France.
Reprod Sci. 2017 Jul;24(7):1062-1069. doi: 10.1177/1933719116678688. Epub 2016 Nov 21.
Our electron microscopy observations demonstrate for the first time that the number of microvilli on the mice oocyte membrane decreases when meiosis progresses from prophase I to metaphase II (MII) stage, and the morphology of the microvilli also changes. Microvilli are significantly shorter and larger on the ovulated oocyte membrane than at the previous stages. Although clathrin vesicles clearly disappear during oocyte maturation, exosome-like vesicles begin to be secreted at the metaphase I stage, more strongly at the MII stage. Multivesicular bodies are visible only at the MII stage. Since several oocyte tetraspanins are involved in the gamete interaction, Cd9 being congregated on the MII oocyte microvilli, we analyzed the effect of tetraspanin deletion on oocyte membrane morphology. The Cd9 and Cd9 Cd81 deletions are associated with a decreased microvilli density on the MII oocyte surface. Microvilli thickness is significantly increased whatever the deleted tetraspanin gene be. Only Cd9 deletion clearly disturbs the vesicular traffic, increasing the number of clathrin and exosome vesicles. Additional investigations are necessary to elucidate how tetraspanins modulate the microvilli morphology, likely in relation with cytoskeleton. The role of oocyte exosomes in gamete adhesion/fusion remains to be further studied.
我们的电子显微镜观察首次表明,当减数分裂从前期I进展到中期II(MII)阶段时,小鼠卵母细胞膜上的微绒毛数量减少,并且微绒毛的形态也发生变化。排卵的卵母细胞膜上的微绒毛比前几个阶段明显更短且更大。尽管在卵母细胞成熟过程中网格蛋白囊泡明显消失,但外泌体样囊泡在中期I阶段开始分泌,在MII阶段分泌更强烈。多囊泡体仅在MII阶段可见。由于几种卵母细胞四跨膜蛋白参与配子相互作用,Cd9聚集在MII卵母细胞微绒毛上,我们分析了四跨膜蛋白缺失对卵母细胞膜形态的影响。Cd9和Cd9 Cd81缺失与MII卵母细胞表面微绒毛密度降低有关。无论缺失的四跨膜蛋白基因是什么,微绒毛厚度都显著增加。只有Cd9缺失明显扰乱囊泡运输,增加网格蛋白和外泌体囊泡的数量。需要进一步研究以阐明四跨膜蛋白如何调节微绒毛形态,这可能与细胞骨架有关。卵母细胞外泌体在配子粘附/融合中的作用仍有待进一步研究。