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Published on: July 30, 2020
Resveratrol-Loaded PLGA Enhanced Vitrified Oocyte Viability through Rab11fip4/Arf5-Mediated Internalization Route
Yixiao Zhu1, Jun Li2, Guizhen Zhou1
1State Key Laboratory of Animal Biotech Breeding, Key Laboratory of Animal Genetics, Breeding and Reproduction of the MARA, National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
None:
Irreversible injury accompanied by oocyte vitrification is still challenging. Recent advances in nanotechnology offer promising therapeutic potential. However, the molecular mechanisms by which nanomaterials protect vitrified oocytes remain elusive. In this study, resveratrol-loaded poly(lactic-co-glycolic acid) nanoparticles (PLGA-RES) were synthesized, and the successful synthesis was confirmed by the results of Fourier transform infrared spectroscopy (FTIR) characterization. Thereafter, the effect of PLGA-RES on the quality of vitrified oocytes was explored. Our results showed that the synthesized PLGA-RES nanoparticles, with an average size of 163.5 nm, demonstrated time- and dose-dependent internalization in mouse oocytes. Transmission electron microscopy (TEM) analysis revealed their distribution around the membrane and perinuclear region. Treatment with PLGA-RES significantly increased the germinal vesicle breakdown (GVBD) rate and reduced the aneuploidy rate in vitrified oocytes. Transcriptome analysis identified 447 upregulated and 185 downregulated mRNAs, with the endocytosis pathway playing a crucial role in PLGA-RES regulation. Short time-series expression miner analysis (STEM) and weighted gene coexpression network analysis (WGCNA) further identified Rab11fip4 as a core gene in the PLGA-RES internalization, which was also confirmed by a molecular docking study. Rab11fip4 was further found to interact with Arf5 via coimmunoprecipitation (co-IP) analysis. Knockdown of Rab11fip4 significantly impaired oocyte maturation and nanoparticles internalization. Further experiments showed that the internalization of PLGA-RES was decreased by endocytic inhibitors, and the Rab11fip4-mediated recycling endosome pathway could protect PLGA-RES from lysosomal degradation. In summary, PLGA-RES could improve vitrified oocyte viability through the endocytic pathway, highlighting the importance of Rab11fip4 in nanoparticle internalization and functional modulation.
