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Impaired Mitochondrial Stress Response due to CLPP Deletion Is Associated with Altered Mitochondrial Dynamics and Increased Apoptosis in Cumulus Cells

  • E Esencan
  • Z Jiang
  • T Wang
  • M Zhang
  • G Soylemez-Imamoglu
  • E SeliEmail author
Original Article

Abstract

Caseinolytic peptidase P (CLPP) plays a central role in mitochondrial unfolded protein response (mtUPR) and is required for maintaining protein homeostasis in the mitochondria. Global germline Clpp deletion causes female infertility and accelerated follicular depletion. In the current study, we aimed to characterize the role of CLPP in cumulus cell function, gene expression, and mitochondrial ultrastructure. We found that mitochondria in Clpp-deficient cumulus cells have a smaller aspect ratio (length/width) and have a larger coverage area (mitochondrial area/cytoplasmic area) under electron microscopy. These ultrastructural changes were accompanied with diminished expression of mitochondrial dynamics genes. RNA sequencing analysis revealed a significant change in genes related to cellular metabolism in Clpp-deficient cumulus cells compared to wild type. In addition, apoptosis and phagosome pathways were significantly affected. Immunofluorescence assessment confirmed increased apoptotic activity and decreased cell proliferation in cumulus oophorus complexes (COCs) of Clpp-deficient mice. Our findings demonstrate that deletion of CLPP results in significant structural and functional changes in cumulus cells and suggests that mtUPR is required for cumulus cell function.

Keywords

CLPP Cumulus cell Mitochondria Mitochondrial unfolded protein response 

Notes

Funding Information

E.S. is a consultant for and receives research funding from the Foundation for Embryonic Competence (FEC).

Supplementary material

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References

  1. 1.
    Eppig JJ, Downs SM. Chemical signals that regulate mammalian oocyte maturation. Biol Reprod. 1984;30(1):1–11.PubMedCrossRefGoogle Scholar
  2. 2.
    Eppig JJ. Maintenance of meiotic arrest and the induction of oocyte maturation in mouse oocyte-granulosa cell complexes developed in vitro from preantral follicles. Biol Reprod. 1991;45(6):824–30.PubMedCrossRefGoogle Scholar
  3. 3.
    Eppig JJ. Mouse oocyte development in vitro with various culture systems. Dev Biol. 1977;60(2):371–88.PubMedCrossRefGoogle Scholar
  4. 4.
    Huang ZW, Wells D. The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod. 2010;16(10):715–25.PubMedCrossRefGoogle Scholar
  5. 5.
    Albertini DF, Combelles CMH, Benecchi E, Carabatsos MJ. Cellular basis for paracrine regulation of ovarian follicle development. Reproduction. 2001;121(5):647–53.PubMedCrossRefGoogle Scholar
  6. 6.
    Dong JW, Albertini DF, Nishimori K, Kumar TR, Lu NF, Matzuk MM. Growth differentiation factor-9 is required during early ovarian folliculogenesis. Nature. 1996;383(6600):531–5.PubMedCrossRefGoogle Scholar
  7. 7.
    Sutton ML, Gilchrist RB, Thompson JG. Effects of in-vivo and in-vitro environments on the metabolism of the cumulus-oocyte complex and its influence on oocyte developmental capacity. Hum Reprod Update. 2003;9(1):35–48.PubMedCrossRefGoogle Scholar
  8. 8.
    Shimada M. Regulation of oocyte meiotic maturation by somatic cells. Reprod Med Biol. 2012;11(4):177–84.PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Hill S, Van Remmen H. Mitochondrial stress signaling in longevity: a new role for mitochondrial function in aging. Redox Biol. 2014;2:936–44.PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Seli E. Mitochondrial DNA as a biomarker for in-vitro fertilization outcome. Curr Opin Obstet Gynecol. 2016;28(3):158–63.PubMedCrossRefPubMedCentralGoogle Scholar
  11. 11.
    Wang T, Zhang M, Jiang Z, Seli E. Mitochondrial dysfunction and ovarian aging. Am J Rep Immunol. 2017;77(5).CrossRefGoogle Scholar
  12. 12.
    Seli E, Wang T, Horvath TL. Mitochondrial unfolded protein response: a stress response with implications for fertility and reproductive aging. Fertil Steril. 2019;111(2):197–204.PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Schinzel R, Dillin A. Endocrine aspects of organelle stress-cell non-autonomous signaling of mitochondria and the ER. Curr Opin Cell Biol. 2015;33:102–10.PubMedCrossRefPubMedCentralGoogle Scholar
  14. 14.
    Zhao Q, Wang J, Levichkin IV, Stasinopoulos S, Ryan MT, Hoogenraad NJ. A mitochondrial specific stress response in mammalian cells. EMBO J. 2002;21(17):4411–9.PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Benedetti C, Haynes CM, Yang Y, Harding HP, Ron D. Ubiquitin-like protein 5 positively regulates chaperone gene expression in the mitochondrial unfolded protein response. Genetics. 2006;174(1):229–39.PubMedPubMedCentralCrossRefGoogle Scholar
  16. 16.
    Haynes CM, Petrova K, Benedetti C, Yang Y, Ron D. ClpP mediates activation of a mitochondrial unfolded protein response in C. elegans. Dev Cell. 2007;13(4):467–80.PubMedCrossRefGoogle Scholar
  17. 17.
    Jensen MB, Jasper H. Mitochondrial proteostasis in the control of aging and longevity. Cell Metab. 2014;20(2):214–25.PubMedPubMedCentralCrossRefGoogle Scholar
  18. 18.
    Aldridge JE, Horibe T, Hoogenraad NJ. Discovery of genes activated by the mitochondrial unfolded protein response (mtUPR) and cognate promoter elements. PLoS One. 2007;2(9):e874.PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Wang T, Babayev E, Jiang Z, et al. Mitochondrial unfolded protein response gene Clpp is required to maintain ovarian follicular reserve during aging, for oocyte competence, and development of pre-implantation embryos. Aging Cell. 2018;17:e12784.PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Durand F, Hoogenraad N. Assessing mitochondrial unfolded protein response in mammalian cells. Methods Mol Biol. 2017;1567:363–78.PubMedCrossRefGoogle Scholar
  21. 21.
    Haynes CM, Yang Y, Blais SP, Neubert TA, Ron D. The matrix peptide exporter HAF-1 signals a mitochondrial UPR by activating the transcription factor ZC376.7 in C. elegans. Mol Cell. 2010;37(4):529–40.PubMedPubMedCentralCrossRefGoogle Scholar
  22. 22.
    Schulz AM, Haynes CM. UPR(mt)-mediated cytoprotection and organismal aging. Biochim Biophys Acta. 2015;1847(11):1448–56.PubMedPubMedCentralCrossRefGoogle Scholar
  23. 23.
    Jenkinson EM, Rehman AU, Walsh T, Clayton-Smith J, Lee K, Morell RJ, et al. Perrault syndrome is caused by recessive mutations in CLPP, encoding a mitochondrial ATP-dependent chambered protease. Am J Hum Genet. 2013;92(4):605–13.PubMedPubMedCentralCrossRefGoogle Scholar
  24. 24.
    Gispert S, Parganlija D, Klinkenberg M, Dröse S, Wittig I, Mittelbronn M, et al. Loss of mitochondrial peptidase Clpp leads to infertility, hearing loss plus growth retardation via accumulation of CLPX, mtDNA and inflammatory factors. Hum Mol Genet. 2013;22(24):4871–87.PubMedCrossRefGoogle Scholar
  25. 25.
    Seli E, Lalioti MD, Flaherty SM, Sakkas D, Terzi N, Steitz JA. An embryonic poly(a)-binding protein (ePAB) is expressed in mouse oocytes and early preimplantation embryos. P Natl Acad Sci USA. 2005;102(2):367–72.CrossRefGoogle Scholar
  26. 26.
    Jiang ZL, Sun JW, Dong H, et al. Transcriptional profiles of bovine in vivo pre-implantation development. BMC Genomics. 2014;15:756.PubMedPubMedCentralCrossRefGoogle Scholar
  27. 27.
    Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and cufflinks. Nat Protoc. 2012;7(3):562–78.PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Trapnell C, Pachter L, Salzberg SL. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics. 2009;25(9):1105–11.PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Takahashi T, Takahashi E, Igarashi H, Tezuka N, Kurachi H. Impact of oxidative stress in aged mouse oocytes on calcium oscillations at fertilization. Mol Reprod Dev. 2003;66(2):143–52.PubMedCrossRefGoogle Scholar
  30. 30.
    Babayev E, Seli E. Oocyte mitochondrial function and reproduction. Curr Opin Obstet Gynecol. 2015;27(3):175–81.PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Biggers JD, Whittingham DG, Donahue RP. The pattern of energy metabolism in the mouse oocyte and zygote. Proc Natl Acad Sci U S A. 1967;58(2):560–7.PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Cipolat S, Martins de Brito O, Dal Zilio B, Scorrano L. OPA1 requires mitofusin 1 to promote mitochondrial fusion. Proc Natl Acad Sci U S A. 2004;101(45):15927–32.PubMedPubMedCentralCrossRefGoogle Scholar
  33. 33.
    Quiros PM, Ramsay AJ, Sala D, et al. Loss of mitochondrial protease OMA1 alters processing of the GTPase OPA1 and causes obesity and defective thermogenesis in mice. EMBO J. 2012;31(9):2117–33.PubMedPubMedCentralCrossRefGoogle Scholar
  34. 34.
    Ito M, Muraki M, Takahashi Y, Imai M, Tsukui T, Yamakawa N, et al. Glutathione S-transferase theta 1 expressed in granulosa cells as a biomarker for oocyte quality in age-related infertility. Fertil Steril. 2008;90(4):1026–35.PubMedCrossRefPubMedCentralGoogle Scholar
  35. 35.
    Host E, Gabrielsen A, Lindenberg S, Smidt-Jensen S. Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injection. Fertil Steril. 2002;77(3):511–5.PubMedCrossRefPubMedCentralGoogle Scholar
  36. 36.
    Corn CM, Hauser-Kronberger C, Moser M, Tews G, Ebner T. Predictive value of cumulus cell apoptosis with regard to blastocyst development of corresponding gametes. Fertil Steril. 2005;84(3):627–33.PubMedCrossRefPubMedCentralGoogle Scholar
  37. 37.
    Babayev E, Wang T, Szigeti-Buck K, Lowther K, Taylor HS, Horvath T, et al. Reproductive aging is associated with changes in oocyte mitochondrial dynamics, function, and mtDNA quantity. Maturitas. 2016;93:121–30.PubMedPubMedCentralCrossRefGoogle Scholar
  38. 38.
    Boczonadi V, Horvath R. Mitochondria: impaired mitochondrial translation in human disease. Int J Biochem Cell Biol. 2014;48:77–84.PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Yang YJ, Zhang YJ, Li YA. Ultrastructure of human oocytes of different maturity stages and the alteration during in vitro maturation. Fertil Steril. 2009;92(1):396.PubMedPubMedCentralGoogle Scholar
  40. 40.
    Soares TR, Reis SD, Pinho BR, Duchen MR, Oliveira JMA. Targeting the proteostasis network in Huntington's disease. Ageing Res Rev. 2019;49:92–103.PubMedPubMedCentralCrossRefGoogle Scholar
  41. 41.
    Chae YC, Caino MC, Lisanti S, Ghosh JC, Dohi T, Danial NN, et al. Control of tumor bioenergetics and survival stress signaling by mitochondrial HSP90s. Cancer Cell. 2012;22(3):331–44.PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Society for Reproductive Investigation 2020

Authors and Affiliations

  • E Esencan
    • 1
  • Z Jiang
    • 1
    • 2
  • T Wang
    • 1
    • 3
  • M Zhang
    • 1
  • G Soylemez-Imamoglu
    • 1
  • E Seli
    • 1
    Email author
  1. 1.Department of Obstetrics, Gynecology and Reproductive SciencesYale School of MedicineNew HavenUSA
  2. 2.AgCenter, School of Animal SciencesLouisiana State UniversityBaton RougeUSA
  3. 3.Foundation for Embryonic CompetenceBasking RidgeUSA

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