Hostname: page-component-848d4c4894-xm8r8 Total loading time: 0 Render date: 2024-06-20T17:24:13.607Z Has data issue: false hasContentIssue false
  • English
  • Français

Mitochondrial cell-free DNA secreted from porcine granulosa cells

Published online by Cambridge University Press:  14 August 2019

Kazuki Kansaku ,
Yasuhisa Munakata ,
Koumei Shirasuna ,
Takehito Kuwayama  and
Hisataka Iwata Open the ORCID record for Hisataka Iwata [Opens in a new window]
Kazuki Kansaku
Affiliation:
Department of Animal Sciences, Tokyo University of Agriculture, Funako 1737, Kanagawa 243-0034, Japan
Yasuhisa Munakata
Affiliation:
Department of Animal Sciences, Tokyo University of Agriculture, Funako 1737, Kanagawa 243-0034, Japan
Koumei Shirasuna
Affiliation:
Department of Animal Sciences, Tokyo University of Agriculture, Funako 1737, Kanagawa 243-0034, Japan
Takehito Kuwayama
Affiliation:
Department of Animal Sciences, Tokyo University of Agriculture, Funako 1737, Kanagawa 243-0034, Japan
Hisataka Iwata*
Affiliation:
Department of Animal Sciences, Tokyo University of Agriculture, Funako 1737, Kanagawa 243-0034, Japan
*
Address for correspondence: Hisataka Iwata. Professor, Department of Animal Science, Tokyo University of Agriculture. Funako 1737, Kanagawa 243–0034, Japan, E-mail: h1iwata@nodai.ac.jp
Get access Rights & Permissions [Opens in a new window]

Summary

Several studies have proposed that cell-free DNA (cfDNA) is a potential biomarker present in follicular fluid (FF) for oocyte quality. Recently we reported that mitochondria-derived cfDNA (mt-cfDNA) closely reflects the amount of cfDNA in FFs. The present study investigated the mechanism regulating mt-cfDNA secretion from porcine granulosa cells. Oocytes and cumulus cell complexes or granulosa cells (GCs) were cultured in maturation medium for 24 or 48 h respectively. Then, nuclear-derived cell-free DNA (n-cfDNA) or mt-cfDNA contents in the spent medium were examined using real-time polymerase chain reaction. When 10 μM of MG132, a proteasome inhibitor, was added to the culture medium, cellular viability of both COCs and GCs decreased and n-cfDNA significantly increased in the culture medium, whereas mt-cfDNA significantly decreased. Supplementation of the culture medium with GW4869, an inhibitor of intracellular vesicle formation, significantly decreased the mt-cfDNA, whereas no effect was observed on n-cfDNA in the medium of both COCs and GCs. Furthermore, the addition of bafilomycin, an inhibitor of autophagy to the culture medium significantly increased mt-cfDNA in the culture medium. After filtration (0.22 μm) and centrifugation (23,000 g), the mt-cfDNA content of the medium decreased significantly. In conclusion, the proteasomal mitochondrial quality control system is upstream of mt-cfDNA secretion and autophagy plays a role in cellular digestion of mitochondrial DNA in the cytoplasm. It is further suggested that dsDNA is enclosed in certain vesicles or associated with small molecules and secreted into the medium.

Keywords

AutophagyCell-free DNAGranulosa cellsMitochondriaOocytes
Type
Research Article
Information
Zygote , Volume 27 , Issue 5 , October 2019 , pp. 272 - 278
Copyright
© Cambridge University Press 2019 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

*

These authors contributed equally.

References

Baixauli, F, López-Otín, C and Mittelbrunn, M (2014) Exosomes and autophagy: coordinated mechanisms for the maintenance of cellular fitness. Front Immunol 5, 403.CrossRefGoogle ScholarPubMed
Bronkhorst, AJ, Wentzel, JF, Aucamp, J, van Dyk, E, du Plessis, L and Pretorius, PJ (2016) Characterization of the cell-free DNA released by cultured cancer cells. Biochim Biophys Acta 1863, 157–65.CrossRefGoogle ScholarPubMed
Chan, NC, Salazar, AM, Pham, AH, Sweredoski, MJ, Kolawa, NJ, Graham, RL, Hess, S and Chan, DC (2011) Broad activation of the ubiquitin-proteasome system by Parkin is critical for mitophagy. Hum Mol Genet 20, 1726–37.CrossRefGoogle ScholarPubMed
Da Broi, MG, Giorgi, VS, Wang, F, Keefe, DL, Albertini, D and Navarro, PA (2018) Influence of follicular fluid and cumulus cells on oocyte quality: clinical implications. J Assist Reprod Genet 35, 735–51.CrossRefGoogle ScholarPubMed
Guan, Y, Zhang, W, Wang, X, Cai, P, Jia, Q and Zhao, W (2017) Cell-free DNA induced apoptosis of granulosa cells by oxidative stress. Clin Chim Acta 473, 213–7.CrossRefGoogle ScholarPubMed
Hayakawa, K, Esposito, E, Wang, X, Terasaki, Y, Liu, Y, Xing, C, Ji, X and Lo, EH (2016) Transfer of mitochondria from astrocytes to neurons after stroke. Nature 535, 551–5.CrossRefGoogle ScholarPubMed
Hennet, ML and Combelles, CM (2012) The antral follicle: a microenvironment for oocyte differentiation. Int J Dev Biol 56, 819–31CrossRefGoogle ScholarPubMed
Kansaku, K, Munakata, Y, Itami, N, Shirasuna, K, Kuwayama, T and Iwata, H (2018) Mitochondrial dysfunction in cumulus–oocyte complexes increases cell-free mitochondrial DNA. J Reprod Dev 64, 261–6.CrossRefGoogle ScholarPubMed
Kosaka, N, Iguchi, H, Yoshioka, Y, Takeshita, F, Matsuki, Y and Ochiya, T (2010) Secretory mechanisms and intercellular transfer of microRNAs in living cells. J Biol Chem 285, 17442–52.CrossRefGoogle ScholarPubMed
Kwon, D, Parkm, E, Sesaki, H and Kang, SJ (2017) Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) suppresses STING-mediated DNA sensing pathway through inducing mitochondrial fission. Biochem Biophys Res Commun 493, 737–43.CrossRefGoogle ScholarPubMed
Mao, Y, Luo, W, Zhang, L, Wu, W, Yuan, L, Xu, H, Song, J, Fujiwara, K, Abe, JI, LeMaire, SA, Wang, XL and Shen, YH (2017) STING-IRF3 triggers endothelial inflammation in response to free fatty acid-induced mitochondrial damage in diet-induced obesity. Arterioscler Thromb Vasc Biol 37, 920–9.CrossRefGoogle ScholarPubMed
Munakata, Y, Ueda, M, Kawahara-Miki, R, Kansaku, K, Itami, N, Shirasuna, K, Kuwayama, T and Iwata, H (2018) Follicular factors determining granulosa cell number and developmental competence of porcine oocytes. J Assist Reprod Genet 35, 1809–19.CrossRefGoogle ScholarPubMed
Munakata, Y, Shirasuna, K, Kuwayama, T and Iwata, H (2019) Cell-free DNA in medium is associated with the maturation ability of in vitro cultured oocytes. J Reprod Dev 65, 171–5.CrossRefGoogle ScholarPubMed
Sato, D, Itami, N, Tasaki, H, Takeo, S, Kuwayama, T and Iwata, H (2014) Relationship between mitochondrial DNA copy number and SIRT1 expression in porcine oocytes. PLoS One 9, e94488.CrossRefGoogle ScholarPubMed
Stigliani, S, Anserini, P, Venturini, PL and Scaruffi, P (2013) Mitochondrial DNA content in embryo culture medium is significantly associated with human embryo fragmentation. Hum Reprod 28, 2652–60.CrossRefGoogle ScholarPubMed
Tanaka, A, Cleland, MM, Xu, S, Narendra, DP, Suen, DF, Karbowski, M and Youle, RJ (2010) Proteasome and p97 mediate mitophagy and degradation of mitofusins induced by Parkin. J Cell Biol 191, 1367–80.CrossRefGoogle ScholarPubMed
Traver, S, Scalici, E, Mullet, T, Molinari, N, Vincens, C, Anahory, T and Hamamah, S (2015) Cell-free DNA in human follicular microenvironment: new prognostic biomarker to predict in vitro fertilization outcomes. PLoS One 10, e0136172.CrossRefGoogle ScholarPubMed
Wang, W, Kong, P, Ma, G, Li, L, Zhu, J, Xia, T, Xie, H, Zhou, W and Wang, S (2017) Characterization of the release and biological significance of cell-free DNA from breast cancer cell lines. Oncotarget 8, 43180–91.Google ScholarPubMed
Wassermann, R, Gulen, MF, Sala, C, Perin, SG, Lou, Y, Rybniker, J, Schmid-Burgk, JL, Schmidt, T, Hornung, V, Cole, ST and Ablasser, A (2015) Mycobacterium tuberculosis differentially activates cGAS- and inflammasome-dependent intracellular immune responses through ESX-1. Cell Host Microbe 17, 799–810.CrossRefGoogle ScholarPubMed
Watson, RO, Bell, SL, MacDuff, DA, Kimmey, JM, Diner, EJ, Olivas, J, Vance, RE, Stallings, CL, Virgin, HW and Cox, JS (2015) The cytosolic sensor cGAS detects Mycobacterium tuberculosis DNA to induce type I interferons and activate autophagy. Cell Host 17, 811–9.CrossRefGoogle ScholarPubMed
Wydooghe, E, Vandaele, L, Heras, S, De Sutter, P, Deforce, D, Peelman, L, De Schauwer, C and Van Soom, A (2017) Autocrine embryotropins revisited: how do embryos communicate with each other in vitro when cultured in groups? Biol Rev Camb Philos Soc 92, 505–20.CrossRefGoogle ScholarPubMed
Yoshii, SR, Kishi, C, Ishihara, N and Mizushima, N (2011) Parkin mediates proteasome-dependent protein degradation and rupture of the outer mitochondrial membrane. J Biol Chem 22, 19630–40.CrossRefGoogle Scholar
Yoshioka, K, Suzuki, C and Onishi, A (2008) Defined system for in vitro production of porcine embryos using a single basic medium. J Reprod Dev 54, 208–13.CrossRefGoogle ScholarPubMed
Zheng, M, Tong, J, Li, WP, Chen, ZJ and Zhang, C (2018) Melatonin concentration in follicular fluid is correlated with antral follicle count (AFC) and in vitro fertilization (IVF) outcomes in women undergoing assisted reproductive technology (ART) procedures. Gynecol Endocrinol 34, 446–50.CrossRefGoogle ScholarPubMed