PUBLICATION
Studies on chilling sensitivity of early stage zebrafish (Danio rerio) ovarian follicles
- Authors
- Tsai, S., Rawson, D.M., and Zhang, T.
- ID
- ZDB-PUB-090302-9
- Date
- 2009
- Source
- Cryobiology 58(3): 279-286 (Journal)
- Registered Authors
- Rawson, David M., Zhang, Tiantian
- Keywords
- Chilling sensitivity, zebrafish ovarian follicle, KCl buffer, L-15 medium, trypan blue, GVBD, methanol
- MeSH Terms
-
- Animals
- Cell Survival
- Cells, Cultured
- Cold Temperature*
- Cryopreservation/methods
- Cryoprotective Agents/pharmacology
- Dimethyl Sulfoxide/pharmacology
- Female
- Methanol/pharmacology
- Ovarian Follicle/growth & development*
- Ovarian Follicle/metabolism
- Time Factors
- Zebrafish/growth & development*
- PubMed
- 19233154 Full text @ Cryobiology
Citation
Tsai, S., Rawson, D.M., and Zhang, T. (2009) Studies on chilling sensitivity of early stage zebrafish (Danio rerio) ovarian follicles. Cryobiology. 58(3):279-286.
Abstract
Cryopreservation of fish gametes is of great importance in aquaculture, conservation and human genomic research. The creation of gamete cryobanks allows the storage of genetic material of targeted species for almost unlimited time periods. Cryopreservation has been successfully applied to fish sperm of many species, but there has been no success with fish embryos and oocytes. One of the obstacles to fish oocyte cryopreservation is their high chilling sensitivity and expecially at subzero temperatures. Although studies on late stage oocyte cryopreservation has been carried out, there have been no reported studies on cryopreservation of early stage ovarian follicles. The aim of this study is to investigate the chilling sensitivity of early stage zebrafish ovarian follicles before developing protocols for their cryopreservation. Experiments were conducted with stage I (primary growth), stage II (cortical alveolus) and stage III (vetillogenesis) ovarian follicles, which were chilled in KCl buffer and L-15 medium for up to 144 h at -1 degrees C in a low temperature bath. Ovarian follicles were also exposed to 2M methanol or 2M DMSO in L-15 medium for up to 168 h at -1 and -5 degrees C respectively. Control follicles were kept at 28 degrees C. Ovarian follicle viability was assessed using trypan blue staining. The results showed that stage I and II ovarian follicles are less sensitive to chilling than stage III follicles. These results were also confirmed following in vitro maturation of the chilled ovarian follicles. The results also showed that L-15 medium is more beneficial than KCl buffer for ovarian follicles at all stages. The presence of both methanol and DMSO reduced chilling sensitivity of ovarian follicles at all stages with methanol being the most effective. The study indicated that stage I and II follicles are less sensitive to chilling than stage III follicles, and that early stage zebrafish ovarian follicles may be better candidates for cryopreservation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping