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    Effect of methanol and Me2SO exposure on mitochondrial activity and distribution in stage III ovarian follicles of zebrafish (Danio rerio).

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    Authors
    Zampolla, Tiziana
    Spikings, Emma
    Zhang, Tiantian
    Rawson, David M.
    Affiliation
    University of Bedfordshire
    Issue Date
    2009-10
    Subjects
    cryoprotectants
    mitochondrial activity
    ovarian follicles
    zebrafish
    cryobiology
    
    Metadata
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    Abstract
    In this study the effect of cryoprotectants that have been shown to be the least toxic to zebrafish ovarian follicles (methanol and Me(2)SO), on mitochondria of stage III ovarian follicles was evaluated. The mitochondrial distributional arrangement, mitochondrial membrane potential, mtDNA copy number, ATP levels and ADP/ATP ratios were assessed following exposure to cryoprotectants for 30 min at room temperature. Results obtained by confocal microscopy showed that 30 min exposure to 2M methanol induced a loss of membrane potential, although viability tests showed no decrease in survival even after 5h post-exposure incubation. Higher concentrations of methanol (3 and 4M) induced not only a decrease in mitochondrial membrane potential but also the loss of mitochondrial distributional arrangement, which suggested a compromised mitochondrial function. Furthermore 3 and 4M treatments resulted in a decrease in viability assessed by Fluorescein diacetate-Propidium iodide (FDA-PI) and in a decrease in mtDNA copy number and ADP/ATP ratio after 5h incubation following methanol exposure, indicating a delayed effect. The use of Me(2)SO, which is considered to be a more toxic CPA to zebrafish ovarian follicles than methanol, caused a decrease in viability and a sustained decrease in ATP levels accompanied by failure to maintain mtDNA copy number within 1h post-exposure incubation. These results indicated that even CPAs that are considered to have no toxicity as determined by Trypan blue (TB) and FDA-PI tests can have a deleterious effect on mitochondrial activity, potentially compromising oocyte growth and embryo development.
    Citation
    Zampolla, T., Spikings, E., Zhang, T., Rawson, D.M. (2009) 'Effect of methanol and Me2SO exposure on mitochondrial activity and distribution in stage III ovarian follicles of zebrafish (Danio rerio)' Cryobiology 59 (2):188-194
    Publisher
    Elsevier
    Journal
    Cryobiology
    URI
    http://hdl.handle.net/10547/228725
    DOI
    10.1016/j.cryobiol.2009.07.002
    PubMed ID
    19595995
    Additional Links
    http://www.sciencedirect.com/science/article/pii/S0011224009000960
    Type
    Article
    Language
    en
    ISSN
    1090-2392
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.cryobiol.2009.07.002
    Scopus Count
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    Cell and Cryobiology Research Group

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