Mitochondrial dysfunction in cumulus-oocyte complexes increases cell-free mitochondrial DNA
| Autor: | Kazuki Kansaku, Hisataka Iwata, Nobuhiko Itami, Takehito Kuwayama, Yasuhisa Munakata, Koumei Shirasuna |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Oocyte Mitochondrial DNA Swine Granulosa cell Mitochondrion DNA Mitochondrial Andrology Cell-free DNA 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Lactate dehydrogenase medicine Animals Cumulus Cells Granulosa Cells 030219 obstetrics & reproductive medicine Chemistry Follicular fluid Mitochondria Follicular Fluid Nuclear DNA 030104 developmental biology medicine.anatomical_structure Apoptosis Oocytes Original Article Female Animal Science and Zoology Cell-Free Nucleic Acids |
| Zdroj: | The Journal of Reproduction and Development |
| ISSN: | 1348-4400 0916-8818 |
| DOI: | 10.1262/jrd.2018-012 |
| Popis: | This study examined the concentration of cell-free mitochondrial DNA (cf-mtDNA) in porcine follicular fluid (FF) and explored whether the cfDNA level in the culture medium could reflect mitochondrial dysfunction in cumulus cell-oocyte complexes (COCs). cfDNA concentration was higher in the fluid of small-sized follicles, compared to that in larger follicles. The length of cfDNA ranged from short (152 bp) to long (1,914 bp) mtDNA in FF, detected by polymerase chain reaction (PCR). cfDNA concentration in FF significantly correlated with the mtDNA copy number in FF but not with the number of one-copy gene (nuclear DNA) in FF. When the COCs were treated with the mitochondrial uncoupler, namely carbonyl cyanide m-chlorophenyl hydrazone (CCCP), for 2 h and incubated for 42 h, subsequent real-time PCR detected significantly higher amount of cf-mtDNA, compared to nuclear cfDNA, in the spent culture medium. The mtDNA number and viability of cumulus cells and oocytes remained unchanged. When the oocytes were denuded from the cumulus cells following CCCP treatment, PCR detected very low levels of cfDNA in the spent culture medium of the denuded oocytes. In contrast, CCCP treatment of granulosa cells significantly increased the amount of cf-mtDNA in the spent culture medium, without any effect on other markers, including survival rate, apoptosis of cumulus cells, and lactate dehydrogenase levels. Thus, cf-mtDNA was present in FF in a wide range of length, and mitochondrial dysfunction in COCs increased the active secretion of cf-mtDNA in the cultural milieu. |
| Databáze: | OpenAIRE |
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