PUBLICATION

Effect of methanol on mitochondrial organization in zebrafish (Danio rerio) ovarian follicles

Authors
Spikings, E., Zampolla, T., Rawson, D., Wang, Y., aned Zhang, T.
ID
ZDB-PUB-110823-19
Date
2012
Source
Theriogenology   77(1): 28-38 (Journal)
Registered Authors
Rawson, David M.
Keywords
mitochondria, cryoprotectant, zebrafish, ovarian follicle
MeSH Terms
  • Adenosine Triphosphate/biosynthesis
  • Animals
  • Biological Transport/drug effects
  • DNA, Mitochondrial/metabolism
  • Electron Transport Complex IV/analysis
  • Electron Transport Complex IV/metabolism
  • Female
  • Immunohistochemistry
  • Methanol/toxicity*
  • Mitochondria/drug effects*
  • Ovarian Follicle/drug effects*
  • Tubulin/analysis
  • Tubulin/metabolism
  • Zebrafish*
PubMed
21855987 Full text @ Theriogenology
Abstract
Successful cryopreservation is usually measured in terms of cell survival. However, there may also be more subtle effects within cells that survive. Previous studies on zebrafish have produced evidence of mitochondrial DNA (mtDNA) damage in cryopreserved embryonic blastomeres and, after exposure to cryoprotectants, alterations in mtDNA replication in embryos and decreased mitochondrial membrane potential, mtDNA and ATP production in ovarian follicles. This study shows that the decreased ATP levels previously observed in stage III zebrafish ovarian follicles exposed to e3 M methanol persisted in those follicles that subsequently developed to stage IV. However, the decreased mtDNA levels were restored in those follicles. In order to determine whether mitochondrial distribution and/or their transport network was affected by the methanol exposure, immunocytochemistry analysis of tubulin and mitochondrial cytochrome c oxidase I (COX-I) was performed, along with phalloidin staining of polymerized actin. Neat arrangements of all proteins were observed in control follicles, with COX-I and tubulin being colocalized near granulosa cell nuclei, while actin formed hexagonal and/or polygonal structures nearer granulosa cell membranes and projected into the oocyte surface. Exposure to methanol (2 to 4 M) disrupted the COX-I and tubulin arrangements and the hexagonal and/or polygonal actin distribution and actin projections into the oocyte. These effects were still observed in those follicles that developed to stage IV, although the severity was reduced. In summary, the disruption to function and distribution of mitochondria in ovarian follicles exposed to > 2 M methanol may be mediated via disruption of the mitochondrial transport system. Some recovery of this disruption may take place after methanol removal and subsequent follicle maturation.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping