Cryopreservation did not affect sperm DNA methylation levels of genes related to fertilization and embryonic development of cynomolgus macaque (Macaca fascicularis)

Bingbing Chen; Zhenwei Lan; Wendell Q Sun; Qinghua Cui; Wei Si

doi:10.1016/j.cryobiol.2022.07.004

Cryopreservation did not affect sperm DNA methylation levels of genes related to fertilization and embryonic development of cynomolgus macaque (Macaca fascicularis)

Cryobiology. 2022 Oct:108:51-56. doi: 10.1016/j.cryobiol.2022.07.004. Epub 2022 Aug 1.

Authors

Bingbing Chen¹, Zhenwei Lan², Wendell Q Sun³, Qinghua Cui⁴, Wei Si⁵

Affiliations

¹ Institute of Biothermal Science and Technology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China; State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China.
² Laboratory of Biochemistry and Molecular Biology, School of Life Sciences, Yunnan University, Yunnan, 650091, China.
³ Institute of Biothermal Science and Technology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China.
⁴ Laboratory of Biochemistry and Molecular Biology, School of Life Sciences, Yunnan University, Yunnan, 650091, China. Electronic address: cuiqinghua@ynu.edu.cn.
⁵ State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China. Electronic address: siw@lpbr.cn.

PMID: 35926569
DOI: 10.1016/j.cryobiol.2022.07.004

Abstract

DNA methylation alters gene expression in numerous biological processes, including embryonic development. It is little known about the effect of cryopreservation on sperm DNA methylation. The present study has investigated whether cryopreservation causes abnormal DNA methylation in cynomolgus macaque sperm for five critical genes that includes the maternally imprinted gene (SNRPN), genes associated with male infertility (HSPA1L, MTHFR) and genes involved in embryonic development (TET3, LZTR1). Our results showed that sperm motility, the percentage of acrosomal integrity, DNA integrity and mitochondrial membrane potential were decreased after cryopreservation either being frozen with penetrating cryoprotectant, glycerol (Gly) or ethylene glycol (EG), compared to fresh sperm (p = 0.000), but the methylation patterns of the five target genes from cynomolgus macaque sperm samples were not affected after cryopreservation as evaluated by the Bisulfite Sequencing PCR (BSP) method. The data indicates that the current protocol for sperm cryopreservation of cynomolgus macaque is safe in terms of DNA methylation levels in these genes related to critical sperm functions.

Keywords: Cynomolgus macaque; DNA methylation; Sperm cryopreservation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cryopreservation* / methods
DNA Methylation
Embryonic Development
Ethylene Glycol
Female
Fertilization
Glycerol
Macaca fascicularis / genetics
Male
Pregnancy
Semen
Semen Preservation* / methods
Semen Preservation* / veterinary
Sperm Motility / genetics
Spermatozoa
snRNP Core Proteins

Substances

snRNP Core Proteins
Ethylene Glycol
Glycerol