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dc.contributor.authorSpikings, Emmaen_GB
dc.contributor.authorZampolla, Tizianaen_GB
dc.contributor.authorRawson, David M.en_GB
dc.contributor.authorWang, Y.en_GB
dc.contributor.authorZhang, Tiantianen_GB
dc.date.accessioned2012-06-11T10:35:33Zen
dc.date.available2012-06-11T10:35:33Zen
dc.date.issued2012-01-01en
dc.identifier.citationSpikings, E., Zampolla, T., Rawson, D.M., Wang, Y., Zhang, T. (2012) 'Effect of methanol on mitochondrial organization in zebrafish (Danio rerio) ovarian follicles' Theriogenology 77 (1):28-38en_GB
dc.identifier.issn1879-3231en
dc.identifier.pmid21855987en
dc.identifier.doi10.1016/j.theriogenology.2011.07.009en
dc.identifier.urihttp://hdl.handle.net/10547/228319en
dc.description.abstractSuccessful 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 ≥3 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.
dc.language.isoenen
dc.publisherElsevieren_GB
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pubmed/21855987en_GB
dc.relation.urlhttp://www.sciencedirect.com/science/article/pii/S0093691X11003281en
dc.subjectmitochondriaen_GB
dc.subjectcryoprotectanten
dc.subjectzebrafishen
dc.subjectovarian follicleen
dc.subjectcryobiologyen
dc.titleEffect of methanol on mitochondrial organization in zebrafish (Danio rerio) ovarian follicles.en
dc.typeArticleen
dc.contributor.departmentUniversity of Bedfordshireen_GB
dc.contributor.departmentChina Agricultural Universityen
dc.identifier.journalTheriogenologyen_GB
html.description.abstractSuccessful 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 ≥3 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.


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