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

  • Tiziana Zampolla
    ,
  • Emma Spikings
    ,
  • Tiantian Zhang
    ,
  • David M. Rawson
  • University of Bedfordshire
Research Output: Contribution to journal Article Peer-review

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.

Publication Information

Output type

Research Output: Contribution to journal Article Peer-review

Original language

English

Pages from-to (Number of pages)

Pages 188-194

Journal (Volume, Issue Number)

Cryobiology (Volume 59, Issue 2)

Publication milestones

  • Accepted/In press - 02/07/2009
  • Published - 11/07/2009

Publication status

Published - 11/07/2009

ISSN

0011-2240

External Publication IDs

  • handle.net: 10547/228725
  • Scopus: 69249209874

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