{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T15:47:09Z","timestamp":1774367229463,"version":"3.50.1"},"reference-count":16,"publisher":"Springer Science and Business Media LLC","issue":"3","license":[{"start":{"date-parts":[[2024,2,10]],"date-time":"2024-02-10T00:00:00Z","timestamp":1707523200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,2,10]],"date-time":"2024-02-10T00:00:00Z","timestamp":1707523200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100015720","name":"Universidad de Extremadura","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100015720","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Vet Res Commun"],"published-print":{"date-parts":[[2024,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The Mitochondrial distribution pattern or MDP in mammalian oocytes serves as an indicator of their cytoplasmic maturity, with a heterogeneous pattern associated with mature cytoplasm. Currently, MDP assessment involves fluorescent labelling of mitochondria followed by visual evaluation, as no quantitative method exists. Our objective was to develop a quantitative approach to assess MDP in mature equine oocytes. Equine oocytes, obtained by ovum pick up (OPU)\u00a0were matured in vitro, and only metaphase II oocytes were used in the study (<jats:italic>n<\/jats:italic>\u2009=\u200956). Following denudation, oocytes were fixed, stained with MitoTracker\u2122 Red CMXRos (50 nM in TCM-199 with Hank\u00b4s salts and 10% FBS) for 15\u00a0min at 38\u00a0\u00b0C, and then incubated with 2.5\u00a0\u00b5g\/ml Hoechst 33342 for 10\u00a0min at 38\u00a0\u00b0C. Confocal microscope images were acquired, and the oocyte\u2019s MDP was visually classified as either homogeneous (HoD; <jats:italic>n<\/jats:italic>\u2009=\u200917) or heterogeneous (HeD; <jats:italic>n<\/jats:italic>\u2009=\u200939). For quantitative analysis, Fiji-ImageJ software was employed. Background subtraction was performed, and a 1-pixel line along the diameter was drawn to calculate the intensity profile. Fluorescence intensities were normalized, and ratios of peripheral to central fluorescence intensity were determined. Student\u00b4s t-test was used for comparations; MDP ratio was (mean\u2009\u00b1\u2009standard error of the mean): 0.8\u2009\u00b1\u20090.02 for HoD and 0.3\u2009\u00b1\u20090.02 for HeD (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001). These results demonstrate concordance between quantitative and qualitative MDP assessment in mature equine oocytes. Our study describes a new approach to quantify mitochondrial distribution pattern and cytoplasmic maturation in mature equine oocytes.<\/jats:p>","DOI":"10.1007\/s11259-024-10325-z","type":"journal-article","created":{"date-parts":[[2024,2,10]],"date-time":"2024-02-10T06:02:06Z","timestamp":1707544926000},"page":"1867-1871","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Description of a new quantitative method to assess mitochondrial distribution pattern in mature equine oocytes"],"prefix":"10.1007","volume":"48","author":[{"given":"Marcos","family":"Luis-Calero","sequence":"first","affiliation":[]},{"given":"Pablo","family":"Fern\u00e1ndez-Hern\u00e1ndez","sequence":"additional","affiliation":[]},{"given":"Jos\u00e9 Manuel","family":"Ortiz-Rodr\u00edguez","sequence":"additional","affiliation":[]},{"given":"Carmen Cristina","family":"Mu\u00f1oz-Garc\u00eda","sequence":"additional","affiliation":[]},{"given":"Isaac","family":"Jardin","sequence":"additional","affiliation":[]},{"given":"Beatriz","family":"Mac\u00edas-Garc\u00eda","sequence":"additional","affiliation":[]},{"given":"Lauro","family":"Gonz\u00e1lez-Fern\u00e1ndez","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2024,2,10]]},"reference":[{"issue":"7","key":"10325_CR1","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1016\/j.theriogenology.2008.12.002","volume":"71","author":"B Ambruosi","year":"2009","unstructured":"Ambruosi B, Lacalandra GM, Iorga AI, De Santis T, Mugnier S, Matarrese R, Goudet G, Dell\u2019Aquila ME (2009) Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: implications on oocyte maturation, fertilization and developmental competence after ICSI. Theriogenology 71(7):1093\u20131104. https:\/\/doi.org\/10.1016\/j.theriogenology.2008.12.002","journal-title":"Theriogenology"},{"issue":"2","key":"10325_CR2","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1093\/humrep\/15.suppl_2.189","volume":"15","author":"BD Bavister","year":"2000","unstructured":"Bavister BD, Squirrell JM (2000) Mitochondrial distribution and function in oocytes and early embryos. Hum Reprod 15(2):189\u2013198. https:\/\/doi.org\/10.1093\/humrep\/15.suppl_2.189","journal-title":"Hum Reprod"},{"issue":"1","key":"10325_CR3","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/1477-7827-3-26","volume":"3","author":"M Caillaud","year":"2005","unstructured":"Caillaud M, Duchamp G, G\u00e9rard N (2005) In vivo effect of interleukin-1beta and interleukin-1RA on oocyte cytoplasmic maturation, ovulation, and early embryonic development in the mare. Reprod Biol Endocrinol 3(1):26. https:\/\/doi.org\/10.1186\/1477-7827-3-26","journal-title":"Reprod Biol Endocrinol"},{"key":"10325_CR4","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.theriogenology.2020.10.036","volume":"60","author":"G Cl\u00e9rico","year":"2021","unstructured":"Cl\u00e9rico G, Taminelli G, Veronesi JC, Polola J, Pagura N, Pinto C, Sansinena M (2021) Mitochondrial function, blastocyst development and live foals born after ICSI of immature vitrified\/warmed equine oocytes matured with or without melatonin. Theriogenology 60:40\u201349. https:\/\/doi.org\/10.1016\/j.theriogenology.2020.10.036","journal-title":"Theriogenology"},{"key":"10325_CR5","doi-asserted-by":"crossref","unstructured":"Fern\u00e1ndez-Hern\u00e1ndez P, Valero-Gonz\u00e1lez M, Fuentes-Romero B, Iglesias-Garc\u00eda M, Ezquerra-Calvo J, Mart\u00edn-Cuervo M, Mac\u00edas-Garc\u00eda B (2023) Resolution of two cases of ovarian abscesses in mares subjected to Ovum pick up. Equine Vet J. Accepted for publication","DOI":"10.1111\/evj.14031"},{"issue":"5","key":"10325_CR6","doi-asserted-by":"publisher","first-page":"836","DOI":"10.1016\/j.theriogenology.2008.10.023","volume":"71","author":"EM Ferreira","year":"2009","unstructured":"Ferreira EM, Vireque AA, Adona PR, Meirelles FV, Ferriani RA, Navarro PAAS (2009) Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence. Theriogenology 71(5):836\u2013848. https:\/\/doi.org\/10.1016\/j.theriogenology.2008.10.023","journal-title":"Theriogenology"},{"key":"10325_CR7","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1111\/rda.12028","volume":"47","author":"R Gonz\u00e1lez","year":"2012","unstructured":"Gonz\u00e1lez R, G\u00f3mez M, Pope C, Brandt Y (2012) Characterization of mitochondrial and actin patterns in cat oocytes and blastocysts. Reprod Domest Anim 47:118\u2013120. https:\/\/doi.org\/10.1111\/rda.12028","journal-title":"Reprod Domest Anim"},{"issue":"2","key":"10325_CR8","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1071\/RD16441","volume":"30","author":"L Gonz\u00e1lez-Fern\u00e1ndez","year":"2018","unstructured":"Gonz\u00e1lez-Fern\u00e1ndez L, S\u00e1nchez-Calabuig MJ, Alves MG, Oliveira PF, Macedo S, Guti\u00e9rrez-Ad\u00e1n A, Rocha A, Mac\u00edas-Garc\u00eda B (2018) Expanded equine cumulus\u2013oocyte complexes exhibit higher meiotic competence and lower glucose consumption than compact cumulus\u2013oocyte complexes. Reprod Fertil Dev 30(2):297. https:\/\/doi.org\/10.1071\/RD16441","journal-title":"Reprod Fertil Dev"},{"issue":"2","key":"10325_CR9","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1111\/rda.13259","volume":"53","author":"K Hinrichs","year":"2018","unstructured":"Hinrichs K (2018) Assisted reproductive techniques in mares. Reprod Domest Anim 53(2):4\u201313. https:\/\/doi.org\/10.1111\/rda.13259","journal-title":"Reprod Domest Anim"},{"issue":"9","key":"10325_CR10","doi-asserted-by":"publisher","first-page":"2484","DOI":"10.3390\/cells10092484","volume":"10","author":"A Kirillova","year":"2021","unstructured":"Kirillova A, Smitz JEJ, Sukhikh GT, Mazunin I (2021) The role of Mitochondria in Oocyte Maturation. Cells 10(9):2484. https:\/\/doi.org\/10.3390\/cells10092484","journal-title":"Cells"},{"key":"10325_CR11","doi-asserted-by":"publisher","first-page":"103097","DOI":"10.1016\/j.jevs.2020.103097","volume":"89","author":"G Lazzari","year":"2020","unstructured":"Lazzari G, Colleoni S, Crotti G, Turini P, Fiorini G, Barandalla M, Landriscina L, Dolci G, Benedetti M, Duchi R, Galli C (2020) Laboratory Production of Equine Embryos. J Equine Vet Sci 89:103097. https:\/\/doi.org\/10.1016\/j.jevs.2020.103097","journal-title":"J Equine Vet Sci"},{"issue":"5","key":"10325_CR12","doi-asserted-by":"publisher","first-page":"1550","DOI":"10.1016\/j.fertnstert.2009.03.050","volume":"93","author":"S Liu","year":"2010","unstructured":"Liu S, Li Y, Gao X, Yan JH, Chen ZJ (2010) Changes in the distribution of mitochondria before and after in vitro maturation of human oocytes and the effect of in vitro maturation on mitochondria distribution. Fertil Steril 93(5):1550\u20131555. https:\/\/doi.org\/10.1016\/j.fertnstert.2009.03.050","journal-title":"Fertil Steril"},{"issue":"1","key":"10325_CR13","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1186\/1477-7827-12-99","volume":"12","author":"NA Martino","year":"2014","unstructured":"Martino NA, Dell\u2019Aquila ME, Filioli Uranio M, Rutigliano L, Nicassio M, Lacalandra GM, Hinrichs K (2014) Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy\/redox potential. Reprod Biol Endocrinol 12(1):99. https:\/\/doi.org\/10.1186\/1477-7827-12-99","journal-title":"Reprod Biol Endocrinol"},{"issue":"1","key":"10325_CR14","doi-asserted-by":"publisher","first-page":"114","DOI":"10.3390\/cancers14010114","volume":"14","author":"J Sanchez-Collado","year":"2021","unstructured":"Sanchez-Collado J, Lopez JJ, Cantonero C, Jardin I, Regod\u00f3n S, Redondo PC, Gordillo J, Smani T, Salido GM, Rosado JA (2021) Orai2 modulates Store-operated Ca2\u2009+\u2009entry and cell cycle progression in breast Cancer cells. Cancers 14(1):114. https:\/\/doi.org\/10.3390\/cancers14010114","journal-title":"Cancers"},{"issue":"2","key":"10325_CR15","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1111\/j.1439-0531.2006.00749.x","volume":"42","author":"H Torner","year":"2007","unstructured":"Torner H, Alm H, Kanitz W, Goellnitz K, Becker F, Poehland R, Bruessow KP, Tuchscherer A (2007) Effect of initial Cumulus morphology on meiotic dynamic and status of Mitochondria in Horse oocytes during IVM. Reprod Domest Anim 42(2):176\u2013183. https:\/\/doi.org\/10.1111\/j.1439-0531.2006.00749.x","journal-title":"Reprod Domest Anim"},{"issue":"3","key":"10325_CR16","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1111\/j.1439-0531.2011.01900.x","volume":"47","author":"XJ Zhuang","year":"2012","unstructured":"Zhuang XJ, Huang Y, Duan YP, Zhang M, Lu YQ, Lu KH (2012) Translocation of active Mitochondria during Buffalo (Bubalus bubalis) oocytes in vitro maturation, fertilization and preimplantation embryo development: Mitochondria distribution in Buffalo Oocyte Maturation, fertilization and embryo development in vitro. Reprod Domest Anim 47(3):443\u2013448. https:\/\/doi.org\/10.1111\/j.1439-0531.2011.01900.x","journal-title":"Reprod Domest Anim"}],"container-title":["Veterinary Research Communications"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11259-024-10325-z.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s11259-024-10325-z\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11259-024-10325-z.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,3]],"date-time":"2024-06-03T11:14:44Z","timestamp":1717413284000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s11259-024-10325-z"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,10]]},"references-count":16,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2024,6]]}},"alternative-id":["10325"],"URL":"https:\/\/doi.org\/10.1007\/s11259-024-10325-z","relation":{},"ISSN":["0165-7380","1573-7446"],"issn-type":[{"value":"0165-7380","type":"print"},{"value":"1573-7446","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,10]]},"assertion":[{"value":"24 November 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 February 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 February 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All experimental procedures were performed according to Institutional, National and European regulations and were approved by the Institutional Animal Care and Use Committee at the University of Extremadura and \u201cJunta de Extremadura\u201d (Ref. MAM\/JSR).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Animal Ethics"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}