The domestic pig as a model for the study of mitochondrial inheritance

  • Dalen Zuidema
  • Peter SutovskyEmail author


Maternal mitochondrial inheritance is a fundamental paradigm within reproductive biology, yet the molecular mechanisms which underlie this process remain poorly understood. The ubiquitin proteasome system (UPS) and branches of the autophagic pathway have been implicated in taking part in the active degradation of sperm mitochondria post-fertilization. Despite this knowledge, there remains much unknown about this process, including the cofactors and substrates involved, as well as the implications of what occurs when these systems of degradation fail. Mitochondrial inheritance research has utilized a variety of animal models. However, one model that is of particular importance, especially when attempting to link mitochondrial inheritance research to humans, is the domestic pig. Pigs offer relatively easy collection of gametes which are similar to those of humans. Furthermore, pigs are physiologically and anatomically more similar to humans than the majority of other model systems available. Porcine in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), and novel cell-free systems are research tools which can be exploited to provide greater insight into the processes behind sperm mitochondrial degradation. In the future studies of mitochondrial inheritance, pigs will likely play a crucial role as an animal model system.


Mitochondria mtDNA Inheritance Ubiquitin Autophagy Proteasome Sperm Oocyte Fertilization 



We thank Professor Rodney Geisert (MU Animal Science) for the image of the porcine female reproductive tract incorporated in Figure 1, and Professor Kathy Timms (MU OBGYN & Women’s Health) for permission to use unpublished data from a joint research project (Figure 2B). We appreciate our colleagues and collaborators, past and present who have been supporting our research on sperm mitophagy. Porcine gametes for our work have been provided reliably by the NIH National Swine Resource and Research Center (NSRRC), University of Missouri.

Funding information

Work in our laboratory, directly pertinent to this manuscript has been funded by Agriculture and Food Research Initiative Competitive Grant no. 2013-67015-20961. Other research in our laboratory relevant to this review has been funded by the National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture (USDA) grant number 2015-67015-23231, grant number 5 R01 HD084353-02 from NIH National Institute of Child and Human Development, and seed funding from the Food for the twenty-first Century Program of the University of Missouri.

Compliance with ethical statements

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent

Not applicable.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.


  1. Al Rawi S, Louvet-Vallee S, Djeddi A, Sachse M, Culetto E, Hajjar C, Boyd L, Legouis R, Galy V (2011) Postfertilization autophagy of sperm organelles prevents paternal mitochondrial DNA transmission. Science 334:1144–1147CrossRefGoogle Scholar
  2. Albertini DF (2011) On the matter of Krogh’s principle. J Assist Reprod Genet 28:1–2CrossRefGoogle Scholar
  3. Antelman J, Manandhar G, Yi YJ, Li R, Whitworth KM, Sutovsky M, Agca C, Prather RS, Sutovsky P (2008) Expression of mitochondrial transcription factor a (TFAM) during porcine gametogenesis and preimplantation embryo development. J Cell Physiol 217:529–543CrossRefGoogle Scholar
  4. Archibald AL, Bolund L, Churcher C, Fredholm M, Groenen MA, Harlizius B, Lee KT, Milan D, Rogers J, Rothschild MF et al (2010) Pig genome sequence--analysis and publication strategy. BMC Genomics 11:438CrossRefGoogle Scholar
  5. Aw WC, Garvin MR, Ballard JWO (2019) Exogenous factors may differentially influence the selective costs of mtDNA mutations. Adv Anat Embryol Cell BiolGoogle Scholar
  6. Bianchi E, Doe B, Goulding D, Wright GJ (2014) Juno is the egg Izumo receptor and is essential for mammalian fertilization. Nature 508:483–487CrossRefGoogle Scholar
  7. Bianchi E, Wright GJ (2015) Cross-species fertilization: the hamster egg receptor, Juno, binds the human sperm ligand, Izumo1. Philos Trans R Soc Lond Ser B Biol Sci 370:20140101CrossRefGoogle Scholar
  8. Braude P, Bolton V, Moore S (1988) Human gene expression first occurs between the four- and eight-cell stages of preimplantation development. Nature 332:459–461CrossRefGoogle Scholar
  9. Cummins JM, Woodall PF (1985) On mammalian sperm dimensions. J Reprod Fertil 75:153–175CrossRefGoogle Scholar
  10. Day BN (2000) Reproductive biotechnologies: current status in porcine reproduction. Anim Reprod Sci 60-61:161–172CrossRefGoogle Scholar
  11. Flach G, Johnson MH, Braude PR, Taylor RA, Bolton VN (1982) The transition from maternal to embryonic control in the 2-cell mouse embryo. EMBO J 1:681–686CrossRefGoogle Scholar
  12. Galen (1586). Galeni Librorum Quarta Classis Apud luntas, Venetijs [Venice] Google Scholar
  13. Giuffra E, Kijas JM, Amarger V, Carlborg O, Jeon JT, Andersson L (2000) The origin of the domestic pig: independent domestication and subsequent introgression. Genetics 154:1785–1791Google Scholar
  14. Glotzer M, Murray AW, Kirschner MW (1991) Cyclin is degraded by the ubiquitin pathway. Nature 349:132–138CrossRefGoogle Scholar
  15. Hao Y, Mathialagan N, Walters E, Mao J, Lai L, Becker D, Li W, Critser J, Prather RS (2006) Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes. Biol Reprod 75:726–733CrossRefGoogle Scholar
  16. Herbert M, Turnbull D (2018) Progress in mitochondrial replacement therapies. Nat Rev Mol Cell Biol 19:71–72CrossRefGoogle Scholar
  17. Jansen RP (2000) Germline passage of mitochondria: quantitative considerations and possible embryological sequelae. Hum Reprod 15(Suppl 2):112–128CrossRefGoogle Scholar
  18. Jensen TW, Mazur MJ, Pettigew JE, Perez-Mendoza VG, Zachary J, Schook LB (2010) A cloned pig model for examining atherosclerosis induced by high fat, high cholesterol diets. Anim Biotechnol 21:179–187CrossRefGoogle Scholar
  19. Jonas E, de Koning DJ (2015) Genomic selection needs to be carefully assessed to meet specific requirements in livestock breeding programs. Front Genet 6:49CrossRefGoogle Scholar
  20. Kaneda H, Hayashi J, Takahama S, Taya C, Lindahl KF, Yonekawa H (1995) Elimination of paternal mitochondrial DNA in intraspecific crosses during early mouse embryogenesis. Proc Natl Acad Sci U S A 92:4542–4546CrossRefGoogle Scholar
  21. Katayama M, Rieke A, Cantley T, Murphy C, Dowell L, Sutovsky P, Day BN (2007) Improved fertilization and embryo development resulting in birth of live piglets after intracytoplasmic sperm injection and in vitro culture in a cysteine-supplemented medium. Theriogenology 67:835–847CrossRefGoogle Scholar
  22. Kerns K, Zigo M, Drobnis EZ, Sutovsky M, Sutovsky P (2018) Zinc ion flux during mammalian sperm capacitation. Nat Commun 9:2061CrossRefGoogle Scholar
  23. Kong BY, Duncan FE, Que EL, Kim AM, O'Halloran TV, Woodruff TK (2014) Maternally-derived zinc transporters ZIP6 and ZIP10 drive the mammalian oocyte-to-egg transition. Mol Hum Reprod 20:1077–1089CrossRefGoogle Scholar
  24. Kramer P, Bressan P (2018) Our (mother’s) mitochondria and our mind. Perspect Psychol Sci 13:88–100CrossRefGoogle Scholar
  25. Kramer P, Bressan P (2019) Mitochondria inspire a lifestyle. Adv Anat Embryol Cell Biol 231Google Scholar
  26. Kuzmuk KN, Shook LB (2011) Pigs as a model for biomedical sciences. The Genetics of the Pig:426–444Google Scholar
  27. Lee CN, Handrow RR, Lenz RW, Ax RL (1985) Interactions of seminal plasma and glycosaminoglycans on acrosome reactions in bovine spermatozoa in vitro. Gamete Res 12:345–355CrossRefGoogle Scholar
  28. Losano JDA, Padin JF, Mendez-Lopez I, Angrimani DSR, Garcia AG, Barnabe VH, Nichi M (2017) The stimulated glycolytic pathway is able to maintain ATP levels and kinetic patterns of bovine epididymal sperm subjected to mitochondrial uncoupling. Oxidative Med Cell Longev 2017:1682393CrossRefGoogle Scholar
  29. Luo S, Valencia CA, Zhang J, Lee NC, Slone J, Gui B, Wang X, Li Z, Dell S, Brown J et al (2018) Biparental inheritance of mitochondrial DNA in humans. Proc Natl Acad Sci U S A 115:13039–13044CrossRefGoogle Scholar
  30. Luo SM, Ge ZJ, Wang ZW, Jiang ZZ, Wang ZB, Ouyang YC, Hou Y, Schatten H, Sun QY (2013) Unique insights into maternal mitochondrial inheritance in mice. Proc Natl Acad Sci U S A 110:13038–13043CrossRefGoogle Scholar
  31. Maddox-Hyttel P, Dinnyes A, Laurincik J, Rath D, Niemann H, Rosenkranz C, Wilmut I (2001) Gene expression during pre- and peri-implantation embryonic development in pigs. Reprod Suppl 58:175–189Google Scholar
  32. Mao J, O’Gorman C, Sutovsky M, Zigo M, Wells KD, Sutovsky P (2018) Ubiquitin A-52 residue ribosomal protein fusion product 1 (Uba52) is essential for preimplantation embryo development. Biol Open 7Google Scholar
  33. Merlet J, Rubio-Pena K, Al Rawi S, and Galy V (2019) Autophagosomal sperm organelle clearance and mtDNA inheritance in C. elegans. Adv Anat Embryol Cell Biol, 1–24Google Scholar
  34. Mio Y, Iwata K, Yumoto K, Kai Y, Sargant HC, Mizoguchi C, Ueda M, Tsuchie Y, Imajo A, Iba Y et al (2012) Possible mechanism of polyspermy block in human oocytes observed by time-lapse cinematography. J Assist Reprod Genet 29:951–956CrossRefGoogle Scholar
  35. Mtango NR, Sutovsky M, Susor A, Zhong Z, Latham KE, Sutovsky P (2012) Essential role of maternal UCHL1 and UCHL3 in fertilization and preimplantation embryo development. J Cell Physiol 227:1592–1603CrossRefGoogle Scholar
  36. Okamura N, Tajima Y, Soejima A, Masuda H, Sugita Y (1985) Sodium bicarbonate in seminal plasma stimulates the motility of mammalian spermatozoa through direct activation of adenylate cyclase. J Biol Chem 260:9699–9705Google Scholar
  37. Peng L, Wen M, Liu Q, Peng J, Tang S, Hong Y, Liu S, Xiao Y (2018) Persistence and transcription of paternal mtDNA dependent on the delivery strategy rather than mitochondria source in fish embryos. Cell Physiol Biochem 47:1898–1908CrossRefGoogle Scholar
  38. Politi Y, Gal L, Kalifa Y, Ravid L, Elazar Z, Arama E (2014) Paternal mitochondrial destruction after fertilization is mediated by a common endocytic and autophagic pathway in Drosophila. Dev Cell 29:305–320CrossRefGoogle Scholar
  39. Prather RS, Day BN (1998) Practical considerations for the in vitro production of pig embryos. Theriogenology 49:23–32CrossRefGoogle Scholar
  40. Pyle A, Hudson G, Wilson IJ, Coxhead J, Smertenko T, Herbert M, Santibanez-Koref M, Chinnery PF (2015) Extreme-depth re-sequencing of mitochondrial DNA finds no evidence of paternal transmission in humans. PLoS Genet 11:e1005040CrossRefGoogle Scholar
  41. Que EL, Bleher R, Duncan FE, Kong BY, Gleber SC, Vogt S, Chen S, Garwin SA, Bayer AR, Dravid VP et al (2015) Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks. Nat Chem 7:130–139CrossRefGoogle Scholar
  42. Renner S, Fehlings C, Herbach N, Hofmann A, von Waldthausen DC, Kessler B, Ulrichs K, Chodnevskaja I, Moskalenko V, Amselgruber W et al (2010) Glucose intolerance and reduced proliferation of pancreatic beta-cells in transgenic pigs with impaired glucose-dependent insulinotropic polypeptide function. Diabetes 59:1228–1238CrossRefGoogle Scholar
  43. Rogers CS, Hao Y, Rokhlina T, Samuel M, Stoltz DA, Li Y, Petroff E, Vermeer DW, Kabel AC, Yan Z et al (2008) Production of CFTR-null and CFTR-DeltaF508 heterozygous pigs by adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer. J Clin Invest 118:1571–1577CrossRefGoogle Scholar
  44. Rojansky R, Cha MY, Chan DC (2016) Elimination of paternal mitochondria in mouse embryos occurs through autophagic degradation dependent on PARKIN and MUL1. Elife 5Google Scholar
  45. Ross JW, Fernandez de Castro JP, Zhao J, Samuel M, Walters E, Rios C, Bray-Ward P, Jones BW, Marc RE, Wang W et al (2012) Generation of an inbred miniature pig model of retinitis pigmentosa. Invest Ophthalmol Vis Sci 53:501–507CrossRefGoogle Scholar
  46. Sato M, Sato K (2011) Degradation of paternal mitochondria by fertilization-triggered autophagy in C. elegans embryos. Science 334:1141–1144CrossRefGoogle Scholar
  47. Schook LB, Beattie C, Beever J, Donovan S, Jamison R, Zuckermann F, Niemi S, Rothschild M, Rutherford M, Smith D (2005a) Swine in biomedical research: creating the building blocks of animal models. Anim Biotechnol 16:183–190CrossRefGoogle Scholar
  48. Schook LB, Beever JE, Rogers J, Humphray S, Archibald A, Chardon P, Milan D, Rohrer G, Eversole K (2005b) Swine genome sequencing consortium (SGSC): a strategic roadmap for sequencing the pig genome. Comp Funct Genomics 6:251–255CrossRefGoogle Scholar
  49. Schwartz M, Vissing J (2002) Paternal inheritance of mitochondrial DNA. N Engl J Med 347:576–580CrossRefGoogle Scholar
  50. Sharpley MS, Marciniak C, Eckel-Mahan K, McManus M, Crimi M, Waymire K, Lin CS, Masubuchi S, Friend N, Koike M et al (2012) Heteroplasmy of mouse mtDNA is genetically unstable and results in altered behavior and cognition. Cell 151:333–343CrossRefGoogle Scholar
  51. Shitara H, Kaneda H, Sato A, Inoue K, Ogura A, Yonekawa H, Hayashi JI (2000) Selective and continuous elimination of mitochondria microinjected into mouse eggs from spermatids, but not from liver cells, occurs throughout embryogenesis. Genetics 156:1277–1284Google Scholar
  52. Shitara H, Kaneda H, Sato A, Iwasaki K, Hayashi J, Taya C, Yonekawa H (2001) Non-invasive visualization of sperm mitochondria behavior in transgenic mice with introduced green fluorescent protein (GFP). FEBS Lett 500:7–11CrossRefGoogle Scholar
  53. Song WH, Sutovsky P (2018) Porcine cell-free system to study mammalian sperm mitophagy. Methods Mol BiolGoogle Scholar
  54. Song WH, Yi YJ, Sutovsky M, Meyers S, Sutovsky P (2016a) The ART and science of sperm mitophagy. Autophagy 12:2510–2511CrossRefGoogle Scholar
  55. Song WH, Yi YJ, Sutovsky M, Meyers S, Sutovsky P (2016b) Autophagy and ubiquitin-proteasome system contribute to sperm mitophagy after mammalian fertilization. Proc Natl Acad Sci U S A 113:E5261–E5270CrossRefGoogle Scholar
  56. Srirattana K, St John JC (2019) Transmission of dysfunctional mitochondrial DNA and its implications for mammalian reproduction. Adv Anat Embryol Cell Biol 231:75–104CrossRefGoogle Scholar
  57. St John JC, Srirattana K, Tsai TS, Sun X (2017) The mitochondrial genome: how it drives fertility. Reprod Fertil Dev 30:118–139CrossRefGoogle Scholar
  58. St John JC, Tsai TS (2018) The association of mitochondrial DNA haplotypes and phenotypic traits in pigs. BMC Genet 19:41CrossRefGoogle Scholar
  59. St John JC, Tsai TS, Cagnone GL (2016) Mitochondrial DNA supplementation as an enhancer of female reproductive capacity. Curr Opin Obstet Gynecol 28:211–216CrossRefGoogle Scholar
  60. Stricker-Krongrad A, Shoemake CR, Pereira ME, Gad SC, Brocksmith D, Bouchard GF (2016) Miniature swine breeds in toxicology and drug safety assessments: what to expect during clinical and pathology evaluations. Toxicol Pathol 44:421–427CrossRefGoogle Scholar
  61. Sutovsky P (2018) Pig overview (male reproduction). In: B Jegou B, skinner MK (eds), Encyclopedia of reproduction second edition vol. 1 Google Scholar
  62. Sutovsky P, Hewitson L, Simerly CR, Tengowski MW, Navara CS, Haavisto A, Schatten G (1996a) Intracytoplasmic sperm injection for Rhesus monkey fertilization results in unusual chromatin, cytoskeletal, and membrane events, but eventually leads to pronuclear development and sperm aster assembly. Hum Reprod 11:1703–1712CrossRefGoogle Scholar
  63. Sutovsky P, McCauley TC, Sutovsky M, Day BN (2003) Early degradation of paternal mitochondria in domestic pig (Sus scrofa) is prevented by selective proteasomal inhibitors lactacystin and MG132. Biol Reprod 68:1793–1800CrossRefGoogle Scholar
  64. Sutovsky P, Moreno RD, Ramalho-Santos J, Dominko T, Simerly C, Schatten G (1999) Ubiquitin tag for sperm mitochondria. Nature 402:371–372CrossRefGoogle Scholar
  65. Sutovsky P, Moreno RD, Ramalho-Santos J, Dominko T, Simerly C, Schatten G (2000) Ubiquitinated sperm mitochondria, selective proteolysis, and the regulation of mitochondrial inheritance in mammalian embryos. Biol Reprod 63:582–590CrossRefGoogle Scholar
  66. Sutovsky P, Navara CS, Schatten G (1996b) Fate of the sperm mitochondria, and the incorporation, conversion, and disassembly of the sperm tail structures during bovine fertilization. Biol Reprod 55:1195–1205CrossRefGoogle Scholar
  67. Sutovsky P, Schatten G (1997) Depletion of glutathione during bovine oocyte maturation reversibly blocks the decondensation of the male pronucleus and pronuclear apposition during fertilization. Biol Reprod 56:1503–1512CrossRefGoogle Scholar
  68. Sutovsky P, Song WH (2017) Post-fertilisation sperm mitophagy: the tale of mitochondrial Eve and Steve. Reprod Fertil Dev 30:56–63CrossRefGoogle Scholar
  69. Sutovsky P, Van Leyen K, McCauley T, Day BN, Sutovsky M (2004) Degradation of paternal mitochondria after fertilization: implications for heteroplasmy, assisted reproductive technologies and mtDNA inheritance. Reprod BioMed Online 8:24–33CrossRefGoogle Scholar
  70. Thompson WE, Ramalho-Santos J, Sutovsky P (2003) Ubiquitination of prohibitin in mammalian sperm mitochondria: possible roles in the regulation of mitochondrial inheritance and sperm quality control. Biol Reprod 69:254–260CrossRefGoogle Scholar
  71. Tsai T, St John JC (2016) The role of mitochondrial DNA copy number, variants, and haplotypes in farm animal developmental outcome. Domest Anim Endocrinol 56(Suppl):S133–S146CrossRefGoogle Scholar
  72. Tsang HG, Rashdan NA, Whitelaw CB, Corcoran BM, Summers KM, MacRae VE (2016) Large animal models of cardiovascular disease. Cell Biochem Funct 34:113–132CrossRefGoogle Scholar
  73. Uh K, Ryu J, Zhang L, Errington J, Machaty Z, Lee K (2019) Development of novel oocyte activation approaches using Zn(2+) chelators in pigs. Theriogenology 125:259–267CrossRefGoogle Scholar
  74. Varea-Sanchez M, Tourmente M, Bastir M, Roldan ER (2016) Unraveling the sperm bauplan: relationships between sperm head morphology and sperm function in rodents. Biol Reprod 95:25CrossRefGoogle Scholar
  75. Vissing J (2019) Paternal comeback in mitochondrial DNA inheritance. Proc Natl Acad Sci U S A 116:1475–1476CrossRefGoogle Scholar
  76. Wei Y, Chiang WC, Sumpter R Jr, Mishra P, Levine B (2017) Prohibitin 2 is an inner mitochondrial membrane mitophagy receptor. Cell 168(224–238):e210Google Scholar
  77. Xia P (2013) Biology of polyspermy in IVF and its clinical indication. Curr Obstet Gynecol Reports 2:226–231CrossRefGoogle Scholar
  78. Yen, N.T., Lin, C.S., Ju, C.C., Wang, S.C., and Huang, M.C. (2007). Mitochondrial DNA polymorphism and determination of effects on reproductive trait in pigs. Reproduction in domestic animals = Zuchthygiene 42, 387-392Google Scholar
  79. Yi YJ, Manandhar G, Sutovsky M, Li R, Jonakova V, Oko R, Park CS, Prather RS, Sutovsky P (2007) Ubiquitin C-terminal hydrolase-activity is involved in sperm acrosomal function and anti-polyspermy defense during porcine fertilization. Biol Reprod 77:780–793CrossRefGoogle Scholar
  80. Zhou Q, Li H, Nakagawa A, Lin JL, Lee ES, Harry BL, Skeen-Gaar RR, Suehiro Y, William D, Mitani S et al (2016) Mitochondrial endonuclease G mediates breakdown of paternal mitochondria upon fertilization. Science 353:394–399CrossRefGoogle Scholar
  81. Zhou Q, Li H, Xue D (2011) Elimination of paternal mitochondria through the lysosomal degradation pathway in C. elegans. Cell Res 21:1662–1669CrossRefGoogle Scholar

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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Animal SciencesUniversity of MissouriColumbiaUSA
  2. 2.Department of Obstetrics, Gynecology and Women’s HealthUniversity of MissouriColumbiaUSA

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