Improvement of Fertilization Capacity and Developmental Ability of Vitrified Bovine Oocytes by JUNO MRNA Microinjection and Cholesterol-Loaded Methyl-β-Cyclodextrin Treatment

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2023 Jan 8

PMID 36614032

Authors

Xi Xu

Tong Hao

Emma Komba

Baigao Yang

Haisheng Hao

Weihua Du

Huabin Zhu

Hang Zhang

Xueming Zhao

Affiliations

Soon will be listed here.

Abstract

Vitrification of oocytes is crucial for embryo biotechnologies, germplasm cryopreservation of endangered and excellent female animals, and the fertility of humans. However, vitrification significantly impairs the fertilization ability of oocytes, which significantly limits its widely used application. JUNO protein, a receptor for Izumo1, is involved in sperm-oocyte fusion and is an indispensable protein for mammalian fertilization, and its abundance is susceptible to vitrification. However, it is still unclear how vitrification reduces the fertilization capacity of bovine oocytes by affecting JUNO protein. This study was designed to investigate the effect of vitrification on the abundance and post-translational modifications of JUNO protein in bovine oocytes. Our results showed that vitrification did not alter the amino acid sequence of JUNO protein in bovine oocytes. Furthermore, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis results showed that vitrification significantly reduced the number and changed the location of disulfide bonds, and increased the number of both phosphorylation and glycosylation sites of JUNO protein in bovine oocytes. Finally, the fertilization capacity and development ability of vitrified oocytes treated with 200 pg JUNO mRNA microinjection and cholesterol-loaded methyl-β-cyclodextrin (CLC/MβCD) were similar to those of fresh oocytes. In conclusion, our results showed that vitrification of bovine oocytes did not alter the protein sequence of JUNO, but induced post-translational modifications and changed protein abundance. Moreover, the fertilization and development ability of vitrified bovine oocytes were improved by the combination treatment of JUNO mRNA microinjection and CLC/MβCD.

References

Qin J, Guo S, Yang J, Qazi I, Pan B, Lv T . Melatonin Promotes Development of Vitrified-Warmed Mouse GV Oocytes, Potentially by Modulating Phosphorylation of Drp1. Front Vet Sci. 2021; 8:752001. PMC: 8497800. DOI: 10.3389/fvets.2021.752001. View

Pitchayapipatkul J, Somfai T, Matoba S, Parnpai R, Nagai T, Geshi M . Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification. Reprod Fertil Dev. 2017; 29(10):2028-2039. DOI: 10.1071/RD16193. View

Sive H, Grainger R, Harland R . Microinjection of Xenopus oocytes. Cold Spring Harb Protoc. 2010; 2010(12):pdb.prot5536. DOI: 10.1101/pdb.prot5536. View

Zhao X, Ren J, Du W, Hao H, Wang D, Qin T . Effect of vitrification on promoter CpG island methylation patterns and expression levels of DNA methyltransferase 1o, histone acetyltransferase 1, and deacetylase 1 in metaphase II mouse oocytes. Fertil Steril. 2013; 100(1):256-61. DOI: 10.1016/j.fertnstert.2013.03.009. View

Wei Y, Lin W, Wang T, Lee W, Li S, Lin F . Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1. J Reprod Dev. 2022; 68(3):198-208. PMC: 9184822. DOI: 10.1262/jrd.2021-128. View

Inoue N . Novel insights into the molecular mechanism of sperm-egg fusion via IZUMO1. J Plant Res. 2016; 130(3):475-478. DOI: 10.1007/s10265-016-0895-z. View

Sudhakaran S, Uppangala S, Salian S, Honguntikar S, Nair R, Kalthur G . Oocytes recovered after ovarian tissue slow freezing have impaired H2AX phosphorylation and functional competence. Reprod Fertil Dev. 2014; 27(8):1242-8. DOI: 10.1071/RD14048. View

Ma Y, Long C, Liu G, Bai H, Ma L, Bai T . WGBS combined with RNA-seq analysis revealed that Dnmt1 affects the methylation modification and gene expression changes during mouse oocyte vitrification. Theriogenology. 2021; 177:11-21. DOI: 10.1016/j.theriogenology.2021.09.032. View

Bodensteiner K, Clay C, Moeller C, Sawyer H . Molecular cloning of the ovine Growth/Differentiation factor-9 gene and expression of growth/differentiation factor-9 in ovine and bovine ovaries. Biol Reprod. 1999; 60(2):381-6. DOI: 10.1095/biolreprod60.2.381. View

10.

Zhang M, Miao Y, Chen Q, Cai M, Dong W, Dai X . BaP exposure causes oocyte meiotic arrest and fertilization failure to weaken female fertility. FASEB J. 2017; 32(1):342-352. DOI: 10.1096/fj.201700514R. View

11.

Brackett B, Oliphant G . Capacitation of rabbit spermatozoa in vitro. Biol Reprod. 1975; 12(2):260-74. DOI: 10.1095/biolreprod12.2.260. View

12.

Sui L, Yan K, Zhang H, Nie J, Yang X, Xu C . Mogroside V Alleviates Oocyte Meiotic Defects and Quality Deterioration in Benzo(a)pyrene-Exposed Mice. Front Pharmacol. 2021; 12:722779. PMC: 8428525. DOI: 10.3389/fphar.2021.722779. View

13.

Miyado K, Yamada G, Yamada S, Hasuwa H, Nakamura Y, Ryu F . Requirement of CD9 on the egg plasma membrane for fertilization. Science. 2000; 287(5451):321-4. DOI: 10.1126/science.287.5451.321. View

14.

Emelyanov A, Fyodorov D . Thioredoxin-dependent disulfide bond reduction is required for protamine eviction from sperm chromatin. Genes Dev. 2016; 30(24):2651-2656. PMC: 5238724. DOI: 10.1101/gad.290916.116. View

15.

Zhao X, Hao H, Du W, Zhao S, Wang H, Wang N . Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes. J Pineal Res. 2015; 60(2):132-41. DOI: 10.1111/jpi.12290. View

16.

Urner F, Sakkas D . Protein phosphorylation in mammalian spermatozoa. Reproduction. 2003; 125(1):17-26. DOI: 10.1530/rep.0.1250017. View

17.

Hao T, Zhang P, Hao H, Du W, Pang Y, Zhao S . The combination treatment of cholesterol-loaded methyl-β-cyclodextrin and methyl-β-cyclodextrin significantly improves the fertilization capacity of vitrified bovine oocytes by protecting fertilization protein JUNO. Reprod Domest Anim. 2021; 56(3):519-530. DOI: 10.1111/rda.13890. View

18.

Liu M, Sun A, Zhao S, Liu H, Ma S, Li M . Resveratrol improves in vitro maturation of oocytes in aged mice and humans. Fertil Steril. 2018; 109(5):900-907. DOI: 10.1016/j.fertnstert.2018.01.020. View

19.

Horvath G, Seidel Jr G . Vitrification of bovine oocytes after treatment with cholesterol-loaded methyl-beta-cyclodextrin. Theriogenology. 2006; 66(4):1026-33. DOI: 10.1016/j.theriogenology.2006.03.004. View

20.

Su J, Wang Y, Li Y, Li R, Li Q, Wu Y . Oxamflatin significantly improves nuclear reprogramming, blastocyst quality, and in vitro development of bovine SCNT embryos. PLoS One. 2011; 6(8):e23805. PMC: 3166058. DOI: 10.1371/journal.pone.0023805. View