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    Impact of cryoprotectants and cryopreservation on metabolic activity and cytoskeleton proteins of zebrafish (Danio rerio) ovarian fragments.

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    Authors
    Zampolla, Tiziana
    Spikings, Emma
    Srirangarajah, Srimathuri
    Rawson, David M.
    Zhang, Tiantian
    Affiliation
    University of Bedfordshire
    Issue Date
    2011-11
    Subjects
    zebrafish
    ovarian fragments
    F-actin
    tubulin
    cytoskeleton
    mitochondria
    cryopreservation
    methanol
    Danio rerio
    
    Metadata
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    Abstract
    Cryopreservation of reproductive cells and tissues of aquatic species offers many benefits to the field of conservation, aquaculture and biomedicine. Although cryopreservation of fish sperm has been successfully achieved, cryopreservation of embryos and oocytes remains unsuccessful. Several studies have been undertaken on cryopreservation of isolated fish ovarian follicles at different stages, although the protocols used lead to a compromised viability. The present study investigates the effect of cryoprotectants and cryopreservation on the viability of ovarian tissues of zebrafish (Danio rerio). The effect of permeating cryoprotectants (CPAs) methanol, dimethyl sulfoxide (DMSO), and ethylene glycol (EG) on ovarian tissues were investigated in a series of toxicity tests. Controlled slow cooling of ovarian tissues using 1M and 4M methanol was also carried out. Ovarian tissue viability was assessed by trypan blue (TB) and fluorescence diacetate (FDA)-propidium iodide (PI) tests. In addition, the effect of methanol exposure and cryopreservation on ovarian follicle ATP level, mitochondria, actin and tubulin distribution were also investigated. Results showed that cryoprotectant toxicity to ovarian fragments increased in the order of methanol, DMSO and EG. The results from controlled slow cooling showed that 1M methanol was more effective than 4M methanol although subsequent cryopreservation induced decreases in ATP levels. Immunocytochemistry and actin staining results showed impacts of cryopreservation on mitochondria and cytoskeleton proteins distribution.
    Citation
    Zampolla, T., Spikings, E. Srirangarajah, S., Rawson, D.M., Zhang, T. (2011) 'Impact of cryoprotectants and cryopreservation on metabolic activity and cytoskeleton proteins of zebrafish (Danio rerio) ovarian fragments' Cryo letters 32 (6):525-536
    Publisher
    Cryo Letters
    Journal
    Cryo letters
    URI
    http://hdl.handle.net/10547/228733
    PubMed ID
    22227713
    Additional Links
    http://www.ncbi.nlm.nih.gov/pubmed/22227713
    Type
    Article
    Language
    en
    ISSN
    0143-2044
    Collections
    Cell and Cryobiology Research Group

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