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

2.50
Hdl Handle:
http://hdl.handle.net/10547/228733
Title:
Impact of cryoprotectants and cryopreservation on metabolic activity and cytoskeleton proteins of zebrafish (Danio rerio) ovarian fragments.
Authors:
Zampolla, Tiziana; Spikings, Emma ( 0000-0001-8387-2098 ) ; Srirangarajah, Srimathuri; Rawson, David M.; Zhang, Tiantian
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.
Affiliation:
LIRANS Institute of Research in the Applied Natural Sciences, University of Bedfordshire, Luton, Bedfordshire, UK.
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
Issue Date:
Nov-2011
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
Appears in Collections:
Cell and Cryobiology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorZampolla, Tizianaen_GB
dc.contributor.authorSpikings, Emmaen_GB
dc.contributor.authorSrirangarajah, Srimathurien_GB
dc.contributor.authorRawson, David M.en_GB
dc.contributor.authorZhang, Tiantianen_GB
dc.date.accessioned2012-06-13T10:59:27Zen
dc.date.available2012-06-13T10:59:27Zen
dc.date.issued2011-11en
dc.identifier.citationZampolla, 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):525-536en_GB
dc.identifier.issn0143-2044en
dc.identifier.pmid22227713en
dc.identifier.urihttp://hdl.handle.net/10547/228733en
dc.description.abstractCryopreservation 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.en_GB
dc.language.isoenen
dc.publisherCryo Lettersen_GB
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pubmed/22227713en_GB
dc.subjectzebrafishen_GB
dc.subjectovarian fragmentsen
dc.subjectF-actinen
dc.subjecttubulinen
dc.subjectcytoskeletonen
dc.subjectmitochondriaen
dc.subjectcryopreservationen
dc.subjectmethanolen
dc.subjectDanio rerioen
dc.subject.meshAnimalsen
dc.subject.meshCryopreservationen
dc.subject.meshCytoskeletal Proteinsen
dc.subject.meshFemaleen
dc.subject.meshImmunohistochemistryen
dc.subject.meshMicroscopy, Confocalen
dc.subject.meshOvaryen
dc.subject.meshZebrafishen
dc.titleImpact of cryoprotectants and cryopreservation on metabolic activity and cytoskeleton proteins of zebrafish (Danio rerio) ovarian fragments.en
dc.typeArticleen
dc.contributor.departmentLIRANS Institute of Research in the Applied Natural Sciences, University of Bedfordshire, Luton, Bedfordshire, UK.en_GB
dc.identifier.journalCryo lettersen_GB

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