PUBLICATION
Impact of cryoprotectants and cryopreservation on metabolic activity and cytoskeleton proteins of zebrafish (Danio rerio) ovarian fragments
- Authors
- Zampolla, T., Spikings, E., Srirangarajah, S., Rawson, D.M., and Zhang, T.
- ID
- ZDB-PUB-120111-32
- Date
- 2011
- Source
- Cryo letters 32(6): 525-536 (Journal)
- Registered Authors
- Rawson, David M., Zhang, Tiantian
- Keywords
- zebrafish, ovarian fragments, F-actin, tubulin, cytoskeleton, mitochondria, cryopreservation, methanol
- MeSH Terms
-
- Animals
- Cryopreservation*
- Cytoskeletal Proteins/metabolism*
- Female
- Immunohistochemistry
- Microscopy, Confocal
- Ovary/metabolism*
- Zebrafish
- PubMed
- 22227713
Citation
Zampolla, T., Spikings, E., Srirangarajah, S., Rawson, D.M., and 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.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping