Succinate dehydrogenase participation in porcine gamete function
E. Breininger A B # , P. Rodriguez A # * , C. Gutnisky A B , G. Alvarez B , M. Satorre A , S. Martinez A , V. Pereyra A , B. Vecchi Galenda A and P. Cetica A BA
B
Handling Editor: Joanna Souza-Fabjan
Abstract
Porcine gametes require energy for the physiological processes that allow fertilisation. Succinate dehydrogenase (SDH) plays a pivotal role in both, the tricarboxylic acid (TCA) cycle and the respiratory chain.
The aim of this work was to study the participation of SDH in the in vitro oocyte maturation, sperm capacitation and acrosome reaction in porcine species.
Cumulus–oocyte complexes (COCs) from abattoir-derived porcine ovaries were collected by aspiration and were incubated in maturation media, with the addition of increasing concentrations (0, 1, 5 and 10 mM) of malonate (a specific inhibitor of SDH). Nuclear maturation and cytoplasmatic maturation were analysed. Semen samples were incubated for 2 h in capacitating medium with 40 mM sodium bicarbonate, as sperm capacitation inducer, and the addition of increasing concentrations of malonate (0, 1, 5 and 10 mM). Sperm capacitation state and true acrosomal reaction were evaluated. SDH activity was determined in sperm and oocyte extracts by the spectrophotometric method.
The addition of 10 mM of malonate decreased both nuclear and cytoplasmic maturation rates (P < 0.05) without affecting COC viability (assessed using fluorescein diacetate). A lower level of capacitation (induced by bicarbonate) and acrosome reaction (induced by follicular fluid) was observed with the addition of 5 mM of malonate (P < 0.05) without affecting motility and viability of sperm at this concentration. The activity of SDH was 0.35 ± 0.1 × 10−5 and 2.37 ± 0.9 × 10−5 U/COC for immature and in vitro matured COC extracts (P < 0.05) respectively, and 0.44 ± 0.16 U/1010 sperm for boar sperm extracts.
In conclusion, because it has been proposed that aerobic and anaerobic metabolic pathways of cells are changed depending on the oxygen availability and the composition of metabolic substrates in their environment, our results suggest that energy obtained through the mitochondrial respiration (TCA cycle and oxidative phosphorylation) is necessary to support oocyte maturation, sperm capacitation and acrosome reaction in the porcine species.
The study of enzymatic activity in gametes is essential for understanding the mechanisms that control the energy production required to achieve successful fertilisation. This knowledge has significant implications for the development of assisted reproductive technologies.
Keywords: acrosome reaction, capacitation, enzyme activity, in vitro maturation, oocyte, porcine, spermatozoa, succinate dehydrogenase.
References
Abeydeera LR, Day BN (1997) Fertilization and subsequent development in vitro of pig oocytes inseminated in a modified tris-buffered medium with frozen-thawed ejaculated spermatozoa. Biology of Reproduction 57, 729-734.
| Crossref | Google Scholar | PubMed |
Abeydeera LR, Wang W-H, Prather RS, Day BN (2001) Effect of incubation temperature on in vitro maturation of porcine oocytes: nuclear maturation, fertilisation and developmental competence. Zygote 9, 331-337.
| Crossref | Google Scholar | PubMed |
Ackrell BAC, Kearney EB, Singer TP (1978) [47] Mammalian succinate dehydrogenase. In ‘Methods in enzymology’. (Eds S Fleischer, L Packer) pp. 466–483. (Academic Press) doi:10.1016/S0076-6879(78)53050-4
Alvarez GM, Dalvit GC, Cetica PD (2012) Influence of the cumulus and gonadotropins on the metabolic profile of porcine cumulus-oocyte complexes during in vitro maturation. Reproduction in Domestic Animals 47(5), 856-864.
| Crossref | Google Scholar | PubMed |
Alvarez GM, Casiró S, Gutnisky C, Dalvit GC, Sutton-McDowall ML, Thompson JG, Cetica PD (2016) Implications of glycolytic and pentose phosphate pathways on the oxidative status and active mitochondria of the porcine oocyte during IVM. Theriogenology 86, 2096-2106.
| Crossref | Google Scholar | PubMed |
Alvarez GM, Expósito MJB, Elia E, Paz D, Morado S, Cetica PD (2019) Effects of gonadotrophins and insulin on glucose uptake in the porcine cumulus–oocyte complex during IVM. Reproduction, Fertility and Development 31, 1353-1359.
| Crossref | Google Scholar | PubMed |
Amoushahi M, Salehnia M (2018) Reactive oxygen species level, mitochondrial transcription factor A gene expression and succinate dehydrogenase activity in metaphase II oocytes derived from in vitro cultured vitrified mouse ovaries. Veterinary Research Forum 9, 145-152.
| Crossref | Google Scholar | PubMed |
Arzondo MN, Caballero JN, Marín-Briggiler CI, et al. (2012) Glass wool filtration of bull cryopreserved semen: a rapid and effective method to obtain a high percentage of functional sperm. Theriogenology 78(1), 201-209.
| Crossref | Google Scholar | PubMed |
Breininger E, Vecchi Galenda BE, Alvarez GM, Gutnisky C, Cetica PD (2014) Phosphofructokinase and malate dehydrogenase participate in the in vitro maturation of porcine oocytes. Reproduction in Domestic Animals 49, 1068-1073.
| Crossref | Google Scholar | PubMed |
Breininger E, Dubois D, Pereyra VE, Rodriguez PC, Satorre MM, Cetica PD (2017) Participation of phosphofructokinase, malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa. Reproduction in Domestic Animals 52, 731-740.
| Crossref | Google Scholar | PubMed |
Brevini Gandolfi TAL, Gandolfi F (2001) The maternal legacy to the embryo: cytoplasmic components and their effects on early development. Theriogenology 55, 1255-1276.
| Crossref | Google Scholar | PubMed |
Brière J-J, Favier J, Ghouzzi VE, Djouadi F, Bénit P, Gimenez A-P, Rustin P (2005) Succinate dehydrogenase deficiency in human. Cellular and Molecular Life Sciences 62, 2317-2324.
| Crossref | Google Scholar | PubMed |
Buccione R, Schroeder AC, Eppig JJ (1990) Interactions between somatic cells and germ cells throughout mammalian oogenesis. Biology of Reproduction 43, 543-547.
| Crossref | Google Scholar | PubMed |
Cetica PD, Pintos LN, Dalvit GC, Beconi MT (1999) Effect of lactate dehydrogenase activity and isoenzyme localization in bovine oocytes and utilization of oxidative substrates on in vitro maturation. Theriogenology 51, 541-550.
| Crossref | Google Scholar | PubMed |
Cetica P, Pintos L, Dalvit G, Beconi M (2003) Involvement of enzymes of amino acid metabolism and tricarboxylic acid cycle in bovine oocyte maturation in vitro. Reproduction 126, 753-763.
| Crossref | Google Scholar | PubMed |
Cimen H, Han M-J, Yang Y, Tong Q, Koc H, Koc EC (2010) Regulation of succinate dehydrogenase activity by SIRT3 in mammalian mitochondria. Biochemistry 49, 304-311.
| Crossref | Google Scholar | PubMed |
Davila MP, Muñoz PM, Bolaños JMG, Stout TAE, Gadella BM, Tapia JA, da Silva CB, Ferrusola CO, Peña FJ (2016) Mitochondrial ATP is required for the maintenance of membrane integrity in stallion spermatozoa, whereas motility requires both glycolysis and oxidative phosphorylation. Reproduction 152, 683-694.
| Crossref | Google Scholar | PubMed |
Desaulniers AT, Cederberg RA, Mills GA, Lents CA, White BR (2017) Production of a gonadotropin-releasing hormone 2 receptor knockdown (GNRHR2 KD) swine line. Transgenic Research 26(4), 567-575.
| Crossref | Google Scholar | PubMed |
du Plessis SS, Agarwal A, Mohanty G, van der Linde M (2015) Oxidative phosphorylation versus glycolysis: what fuel do spermatozoa use? Asian Journal of Andrology 17, 230-235.
| Crossref | Google Scholar |
Eppig JJ, Hosoe M, O’Brien MJ, Pendola FM, Requena A, Watanabe S (2000) Conditions that affect acquisition of developmental competence by mouse oocytes in vitro: FSH, insulin, glucose and ascorbic acid. Molecular and Cellular Endocrinology 163, 109-116.
| Crossref | Google Scholar | PubMed |
Ferramosca A, Zara V (2014) Bioenergetics of mammalian sperm capacitation. BioMed Research International 2014, 902953.
| Crossref | Google Scholar |
Gohil VM, Sheth SA, Nilsson R, Wojtovich AP, Lee JH, Perocchi F, Chen W, Clish CB, Ayata C, Brookes PS, Mootha VK (2010) Nutrient-sensitized screening for drugs that shift energy metabolism from mitochondrial respiration to glycolysis. Nature Biotechnology 28, 249-255.
| Crossref | Google Scholar | PubMed |
Gutnisky C, Morado S, Dalvit GC, Thompson JG, Cetica PD (2013) Glycolytic pathway activity: effect on IVM and oxidative metabolism of bovine oocytes. Reproduction, Fertility and Development 25(7), 1026-1035.
| Crossref | Google Scholar | PubMed |
Hajjawi OS (2011) Succinate dehydrogenase: assembly, regulation and role in human disease. European Journal of Scientific Research 51, 133-142.
| Google Scholar |
Ho H-C, Granish KA, Suarez SS (2002) Hyperactivated motility of bull sperm is triggered at the axoneme by Ca2+ and not cAMP. Developmental Biology 250(1), 208-217.
| Crossref | Google Scholar | PubMed |
Hoppe RW, Bavister BD (1984) Evaluation of the fluorescein diacetate (FDA) vital dye viability test with hamster and bovine embryos. Animal Reproduction Science 6, 323-335.
| Crossref | Google Scholar |
Jones AR (1997) Metabolism of lactate by mature boar spermatozoa. Reproduction, Fertility and Development 9, 227-232.
| Crossref | Google Scholar | PubMed |
Khurana NK, Niemann H (2000) Effects of oocyte quality, oxygen tension, embryo density, cumulus cells and energy substrates on cleavage and morula/blastocyst formation of bovine embryos. Theriogenology 54, 741-756.
| Crossref | Google Scholar | PubMed |
Krebs HA, Kornberg HL (1957) ‘Energy transformations in living matter.’ (Springer) doi:10.1007/978-3-642-86577-0
Krisher RL (2004) The effect of oocyte quality on development. Journal of Animal Science 82((E-Suppl.)), E14-E23.
| Crossref | Google Scholar |
Krisher RL, Brad AM, Herrick JR, Sparman ML, Swain JE (2007) A comparative analysis of metabolism and viability in porcine oocytes during in vitro maturation. Animal Reproduction Science 98, 72-96.
| Crossref | Google Scholar | PubMed |
Leese HJ, Barton AM (1984) Pyruvate and glucose uptake by mouse ova and preimplantation embryos. Journal of Reproduction and Fertility 72, 9-13.
| Crossref | Google Scholar | PubMed |
Lodish MB, Adams KT, Huynh TT, Prodanov T, Ling A, Chen C, Shusterman S, Jimenez C, Merino M, Hughes M, Cradic KW, Milosevic D, Singh RJ, Stratakis CA, Pacak K (2010) Succinate dehydrogenase gene mutations are strongly associated with paraganglioma of the organ of Zuckerkandl. Endocrine-Related Cancer 17, 581-588.
| Crossref | Google Scholar | PubMed |
Marin S, Chiang K, Bassilian S, Lee W-NP, Boros LG, Fernández-Novell JM, Centelles JJ, Medrano A, Rodriguez-Gil JE, Cascante M (2003) Metabolic strategy of boar spermatozoa revealed by a metabolomic characterization. FEBS Letters 554, 342-346.
| Crossref | Google Scholar | PubMed |
Nascimento JM, Shi LZ, Tam J, Chandsawangbhuwana C, Durrant B, Botvinick EL, Berns MW (2008) Comparison of glycolysis and oxidative phosphorylation as energy sources for mammalian sperm motility, using the combination of fluorescence imaging, laser tweezers, and real-time automated tracking and trapping. Journal of Cellular Physiology 217, 745-751.
| Crossref | Google Scholar | PubMed |
O’Flaherty C, Beconi M, Beorlegui N (1997) Effect of natural antioxidants, superoxide dismutase and hydrogen peroxide on capacitation of frozen-thawed bull spermatozoa. Andrologia 29, 269-275.
| Crossref | Google Scholar | PubMed |
Pascual ML, Cebrian-Perez JA, Lopez-Perez MJ, Muino-Blanco T (1996) Short-term inhibition of the energy metabolism affects motility but not surface properties of sperm cells. Bioscience Reports 16, 35-40.
| Crossref | Google Scholar | PubMed |
Pintado B, de la Fuente J, Roldan ER (2000) Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability. Journal of Reproduction and Fertility 118, 145-152.
| Crossref | Google Scholar | PubMed |
Piomboni P, Focarelli R, Stendardi A, Ferramosca A, Zara V (2012) The role of mitochondria in energy production for human sperm motility. International Journal of Andrology 35, 109-124.
| Crossref | Google Scholar | PubMed |
Potter M, Newport E, Morten KJ (2016) The Warburg effect: 80 years on. Biochemical Society Transactions 44, 1499-1505.
| Crossref | Google Scholar |
Ramió-Lluch L, Fernández-Novell JM, Peña A, Colás C, Cebrián-Pérez JA, Muiño-Blanco T, Ramírez A, Concha II, Rigau T, Rodríguez-Gil JE (2011) ‘In vitro’ capacitation and acrosome reaction are concomitant with specific changes in mitochondrial activity in boar sperm: evidence for a nucleated mitochondrial activation and for the existence of a capacitation-sensitive subpopulational structure. Reproduction in Domestic Animals 46, 664-673.
| Crossref | Google Scholar | PubMed |
Rodriguez P, Duarte J, Duprat MP, Pereyra V, Satorre M, Cetica P, Breininger E (2020) Participation of lactate dehydrogenase in capacitation and acrosome reaction of fresh and cryopreserved, with or without α-tocopherol, boar sperm. Animal Science Papers and Reports 38(1), 91-100.
| Google Scholar |
Rodríguez-Gil JE, Bonet S (2016) Current knowledge on boar sperm metabolism: comparison with other mammalian species. Theriogenology 85, 4-11.
| Crossref | Google Scholar | PubMed |
Rodríguez-Gil JE, Estrada E (2013) Artificial insemination in boar reproduction. In ‘Boar reproduction: fundamentals and new biotechnological trends’. (Eds S Bonet, I Casas, W Holt, M Yeste) pp. 589–607. (Springer: Berlin, Germany) doi:10.1007/978-3-642-35049-8_12
Ruiz-Pesini E, Diez C, Lapeña AC, Pérez-Martos A, Montoya J, Alvarez E, Arenas J, López-Pérez MJ (1998) Correlation of sperm motility with mitochondrial enzymatic activities. Clinical Chemistry 44, 1616-1620.
| Crossref | Google Scholar | PubMed |
Satorre MM, Breininger E, Beconi MT, Beorlegui NB (2009) Protein tyrosine phosphorylation under capacitating conditions in porcine fresh spermatozoa and sperm cryopreserved with and without alpha tocopherol. Andrologia 41, 184-192.
| Crossref | Google Scholar | PubMed |
Satorre MM, Breininger E, Cetica PD, Córdoba M (2018) Relation between respiratory activity and sperm parameters in boar spermatozoa cryopreserved with alpha-tocopherol and selected by Sephadex. Reproduction in Domestic Animals 53, 979-985.
| Crossref | Google Scholar | PubMed |
Sirard M-A, Richard F, Blondin P, Robert C (2006) Contribution of the oocyte to embryo quality. Theriogenology 65, 126-136.
| Crossref | Google Scholar | PubMed |
Stival C, Puga Molina LdC, Paudel B, Buffone MG, Visconti PE, Krapf D (2016) Sperm capacitation and acrosome reaction in mammalian sperm. In ‘Sperm acrosome biogenesis and function during fertilization. Advances in Anatomy, Embryology and Cell Biology. Vol. 220’. (Ed. M Buffone) pp. 93–106. (Springer) doi:10.1007/978-3-319-30567-7_5
Sturmey RG, Leese HJ (2003) Energy metabolism in pig oocytes and early embryos. Reproduction 126, 197-204.
| Crossref | Google Scholar | PubMed |
Sutovský P, Fléchon JE, Fléchon B, Motlik J, Peynot N, Chesné P, Heyman Y (1993) Dynamic changes of gap junctions and cytoskeleton during in vitro culture of cattle oocyte cumulus complexes. Biology of Reproduction 49, 1277-1287.
| Crossref | Google Scholar | PubMed |
Sutton ML, Cetica PD, Beconi MT, Kind KL, Gilchrist RB, Thompson JG (2003) Influence of oocyte-secreted factors and culture duration on the metabolic activity of bovine cumulus cell complexes. Reproduction 126, 27-34.
| Crossref | Google Scholar | PubMed |
Tarazona AM, Rodríguez JI, Restrepo LF, Olivera-Angel M (2006) Mitochondrial activity, distribution and segregation in bovine oocytes and in embryos produced in vitro. Reproduction in Domestic Animals 41, 5-11.
| Crossref | Google Scholar | PubMed |
Trimarchi JR, Liu L, Porterfield DM, Smith PJS, Keefe DL (2000) Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos. Biology of Reproduction 62, 1866-1874.
| Crossref | Google Scholar | PubMed |
Wang WH, Abeydeera LR, Fraser LR, Niwa K (1995) Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen-thawed ejaculated boar spermatozoa incubated in a protein-free chemically defined medium. Journal of Reproduction and Fertility 104, 305-313.
| Crossref | Google Scholar | PubMed |
Zhu Z, Li R, Wang L, Zheng Y, Hoque SAM, Lv Y, Zeng W (2019) Glycogen synthase kinase-3 regulates sperm motility and acrosome reaction via affecting energy metabolism in goats. Frontiers in Physiology 10, 968.
| Crossref | Google Scholar | PubMed |