Oocyte mitochondrial bioenergy potential and oxidative stress: within-/between-subject, in vivo versus in vitro maturation, and age-related variations.

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Presentation transcript:

Oocyte mitochondrial bioenergy potential and oxidative stress: within-/between-subject, in vivo versus in vitro maturation, and age-related variations in a sheep model Nicola Antonio Martino, B.Sc., Giovanni Michele Lacalandra, D.M.V., Manuel Filioli Uranio, B.Sc., Barbara Ambruosi, Ph.D., Michele Caira, D.M.V., Fabio Silvestre, Ph.D., Flavia Pizzi, Ph.D., Salvatore Desantis, B.Sc., Gianluca Accogli, B.Sc., Maria Elena Dell’Aquila, Ph.D. Fertility and Sterility Volume 97, Issue 3, Pages 720-728.e1 (March 2012) DOI: 10.1016/j.fertnstert.2011.12.014 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Mitochondrial (mt) distribution pattern and intracellular reactive oxygen species (ROS) localization in sheep oocytes matured in vivo (A and C) or in vitro (B and D). For each oocyte, corresponding bright-field (A1, B1, C1, D1), UV light (A2, B2, C2, D2), confocal images showing mt distribution pattern (A3, B3, C3, D3), ROS localization (A4, B4, C4, D4), mt-ROS merge (A5, B5, C5, D5), and colocalization scatterplot graph (A6, B6, C6, D6) are shown. Images are representative of in vivo (A and C) and in vitro (B and D) matured oocytes with heterogeneous (pericortical/perinuclear, A and B) or homogeneous (small aggregates, C and D) mt distribution pattern. No differences in mt cluster/aggregate size was noticed between in vivo and in vitro matured oocytes. Scale bar represents 40 μm. Fertility and Sterility 2012 97, 720-728.e1DOI: (10.1016/j.fertnstert.2011.12.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Mean ± SD of mitochondrial (mt) distribution patterns and activity, intracellular reactive oxygen species (ROS) levels, and mt-ROS colocalization of sheep oocytes matured in vivo or in vitro. (A) Rates of oocytes showing pericortical/perinuclear (P/P) or small granules (SA) mt pattern. Numbers of analyzed oocytes per group are indicated above the histograms. (B, C) Mitochondrial activity and ROS levels are expressed as (B) Mitotracker Orange CMTM Ros and (C) 2′,7′-dichlorodihydrofluorescein diacetate (DCDHFDA) fluorescence intensity in arbitrary densitometric units (ADU). (D) Pearson correlation coefficients of Mitotracker Orange CMTM Ros and DCDHFDA fluorescent labeling of in vivo versus IVM MIIs. A: chi-square test; B–D: Student t test; ∗P<.05; ∗∗P<.001. Fertility and Sterility 2012 97, 720-728.e1DOI: (10.1016/j.fertnstert.2011.12.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Lectin histochemistry of the oviductal ampullar tract in an hormonally stimulated sheep. Light micrographs of sheep oviductal ampulla stained with (A) Maackia amurensis lectin, (B and C) KOH-sialidase–peanut agglutinin (PNA), and (D and E) Sambucus nigra agglutinin (SNA). Compared with the untreated sheep (B, D), the ampullar epithelium of the hormonally treated sheep displayed a more continuous reactivity with KOH-sialidase-PNA in the luminal surface (C) and higher reactivity with SNA in the supranuclear region of nonciliated cells (E). m = muscular wall; mf = mucosal folds; arrows: luminal surface; arrowheads: supranuclear region of nonciliated cells. Scale bars: A = 500 μm; B–E = 80 μm. Fertility and Sterility 2012 97, 720-728.e1DOI: (10.1016/j.fertnstert.2011.12.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions