Age-related changes in DNA methylation levels at CpG sites in bull spermatozoa and in vitro fertilization-derived blastocyst-stage embryos revealed by combined bisulfite restriction analysis

Autor: Masahiro Kaneda, Kumiko Takeda, Ken Iwao, Hiromichi Adachi, Akira Imai, Shinya Watanabe, Yoichiro Hoshino, Satoshi Akagi, Eiji Kobayashi, Kagetomo Nishino
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Male
Aging
Restriction Mapping
Embryonic Development
Semen
Fertilization in Vitro
Biology
Andrology
Embryo Culture Techniques
03 medical and health sciences
0302 clinical medicine
Human fertilization
Combined bisulfite restriction analysis
medicine
Animals
Combinatorial Chemistry Techniques
Humans
Sulfites
Blastocyst
In vitro fertilization (IVF) embryos
Cells
Cultured
030304 developmental biology
Oligonucleotide Array Sequence Analysis
Combined bisulfite restriction analysis (COBRA)
0303 health sciences
030219 obstetrics & reproductive medicine
Bull spermatozoa
DNA methylation
urogenital system
Age-related changes
Age Factors
Embryo
Methylation
Sequence Analysis
DNA
Embryo
Mammalian
Spermatozoa
medicine.anatomical_structure
CpG site
Animal Science and Zoology
Original Article
Cattle
CpG Islands
Female
Zdroj: The Journal of Reproduction and Development
SC10201910030001
NARO成果DBa
ISSN: 1348-4400
0916-8818
DOI: 10.1262/jrd.2018-146
Popis: Age-associated methylation changes in genomic DNA have been recently reported in spermatozoa, and these changes can contribute to decline in fertility. In a previous study, we analyzed the genome-wide DNA methylation profiles of bull spermatozoa using a human DNA methylation microarray and identified one CpG site (CpG-1) that potentially reflects age-related methylation changes. In the present study, cryopreserved semen samples from a Japanese Black bull were collected at five different ages, which were referred to as JD1-5: 14, 19, 28, 54, and 162 months, respectively, and were used for genome-wide DNA methylation analysis and in vitro fertilization (IVF). Distinct age-related changes in methylation profiles were observed, and 77 CpG sites were found to be differently methylated between young and adult samples (JD1-2 vs. JD4-5). Using combined bisulfite restriction analysis (COBRA), nine CpG sites (including CpG-1) were confirmed to exhibit significant differences in their age-dependent methylation levels. Eight CpG sites showed an age-dependent increase in their methylation levels, whereas only one site showed age-dependent hypomethylation; in particular, these changes in methylation levels occurred rapidly at a young age. COBRA revealed low methylation levels in some CpG regions in the majority of the IVF blastocyst-stage embryos derived from spermatozoa at JD2-5. Interestingly, bulls with different ages did not show differences in their methylation levels. In conclusion, our findings indicated that methylation levels at nine CpG sites in spermatozoa changed with increasing age and that some CpG regions were demethylated after fertilization. Further studies are required to determine whether age-dependent different methylation levels in bull spermatozoa can affect fertility.
Databáze: OpenAIRE
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