Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development.

2.50
Hdl Handle:
http://hdl.handle.net/10547/228294
Title:
Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development.
Authors:
Spikings, Emma ( 0000-0001-8387-2098 ) ; Alderson, Jon; St John, Justin C.
Abstract:
Cellular ATP is mainly generated through mitochondrial oxidative phosphorylation, which is dependent on mitochondrial DNA (mtDNA). We have previously demonstrated the importance of oocyte mtDNA for porcine and human fertilization. However, the role of nuclear-encoded mitochondrial replication factors during oocyte and embryo development is not yet understood. We have analyzed two key factors, mitochondrial transcription factor A (TFAM) and polymerase gamma (POLG), to determine their role in oocyte and early embryo development. Competent and incompetent oocytes, as determined by brilliant cresyl blue (BCB) dye, were assessed intermittently during the maturation process for TFAM and POLG mRNA using real-time RT-PCR, for TFAM and POLG protein using immunocytochemistry, and for mtDNA copy number using real-time PCR. Analysis was also carried out following treatment of maturing oocytes with the mtDNA replication inhibitor, 2',3'-dideoxycytidine (ddC). Following in vitro fertilization, preimplantation embryos were also analyzed. Despite increased levels of TFAM and POLG mRNA and protein at the four-cell stage, no increase in mtDNA copy number was observed in early preimplantation development. To compensate for this, mtDNA appeared to be replicated during oocyte maturation. However, significant differences in nuclear-encoded regulatory protein expression were observed between BCB(+) and BCB(-) oocytes and between untreated oocytes and those treated with ddC. These changes resulted in delayed mtDNA replication, which correlated to reduced fertilization and embryonic development. We therefore conclude that adherence to the regulation of the timing of mtDNA replication during oocyte maturation is essential for successful embryonic development.
Affiliation:
University of Birmingham
Citation:
Spikings, E., Alderson, J., St John, J.C. (2007) 'Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development' Biology of reproduction 76 (2):327-35
Publisher:
Society for the Study of Reproduction
Journal:
Biology of reproduction
Issue Date:
Feb-2007
URI:
http://hdl.handle.net/10547/228294
DOI:
10.1095/biolreprod.106.054536
PubMed ID:
17035641
Additional Links:
http://www.ncbi.nlm.nih.gov/pubmed/17035641; http://www.biolreprod.org/content/76/2/327.long
Type:
Article
Language:
en
ISSN:
1529-7268
Appears in Collections:
Cell and Cryobiology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorSpikings, Emmaen_GB
dc.contributor.authorAlderson, Jonen_GB
dc.contributor.authorSt John, Justin C.en_GB
dc.date.accessioned2012-06-11T08:30:45Zen
dc.date.available2012-06-11T08:30:45Zen
dc.date.issued2007-02en
dc.identifier.citationSpikings, E., Alderson, J., St John, J.C. (2007) 'Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development' Biology of reproduction 76 (2):327-35en_GB
dc.identifier.issn1529-7268en
dc.identifier.pmid17035641en
dc.identifier.doi10.1095/biolreprod.106.054536en
dc.identifier.urihttp://hdl.handle.net/10547/228294en
dc.description.abstractCellular ATP is mainly generated through mitochondrial oxidative phosphorylation, which is dependent on mitochondrial DNA (mtDNA). We have previously demonstrated the importance of oocyte mtDNA for porcine and human fertilization. However, the role of nuclear-encoded mitochondrial replication factors during oocyte and embryo development is not yet understood. We have analyzed two key factors, mitochondrial transcription factor A (TFAM) and polymerase gamma (POLG), to determine their role in oocyte and early embryo development. Competent and incompetent oocytes, as determined by brilliant cresyl blue (BCB) dye, were assessed intermittently during the maturation process for TFAM and POLG mRNA using real-time RT-PCR, for TFAM and POLG protein using immunocytochemistry, and for mtDNA copy number using real-time PCR. Analysis was also carried out following treatment of maturing oocytes with the mtDNA replication inhibitor, 2',3'-dideoxycytidine (ddC). Following in vitro fertilization, preimplantation embryos were also analyzed. Despite increased levels of TFAM and POLG mRNA and protein at the four-cell stage, no increase in mtDNA copy number was observed in early preimplantation development. To compensate for this, mtDNA appeared to be replicated during oocyte maturation. However, significant differences in nuclear-encoded regulatory protein expression were observed between BCB(+) and BCB(-) oocytes and between untreated oocytes and those treated with ddC. These changes resulted in delayed mtDNA replication, which correlated to reduced fertilization and embryonic development. We therefore conclude that adherence to the regulation of the timing of mtDNA replication during oocyte maturation is essential for successful embryonic development.en_GB
dc.language.isoenen
dc.publisherSociety for the Study of Reproductionen_GB
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pubmed/17035641en_GB
dc.relation.urlhttp://www.biolreprod.org/content/76/2/327.longen
dc.rightsArchived with thanks to Biology of reproductionen_GB
dc.subjectembryoen_GB
dc.subjectfertilizationen
dc.subjectgene regulationen
dc.subjectoocyte developmenten
dc.subject.meshAnimalsen
dc.subject.meshBlastocysten
dc.subject.meshCell Agingen
dc.subject.meshCells, Cultureden
dc.subject.meshColoring Agentsen
dc.subject.meshDNA Replicationen
dc.subject.meshDNA, Mitochondrialen
dc.subject.meshDNA-Directed DNA Polymeraseen
dc.subject.meshEmbryo Culture Techniquesen
dc.subject.meshEmbryonic Developmenten
dc.subject.meshFertilizationen
dc.subject.meshFertilization in Vitroen
dc.subject.meshGene Dosageen
dc.subject.meshImmunohistochemistryen
dc.subject.meshOocytesen
dc.subject.meshOxazinesen
dc.subject.meshRNA, Messengeren
dc.subject.meshReverse Transcriptase Inhibitorsen
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen
dc.subject.meshStaining and Labelingen
dc.subject.meshSwineen
dc.subject.meshTime Factorsen
dc.subject.meshTranscription Factorsen
dc.subject.meshZalcitabineen
dc.titleRegulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development.en
dc.typeArticleen
dc.contributor.departmentUniversity of Birminghamen_GB
dc.identifier.journalBiology of reproductionen_GB

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