Journal
ZYGOTE
Volume 8, Issue 3, Pages 209-215Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0967199400001003
Keywords
mitochondria; oocyte; quantification; rapid cycling; real-time PCR
Funding
- NCRR NIH HHS [R21 RR021881-03, R21 RR021881-02, R21 RR021881-01, R21 RR021881-04] Funding Source: Medline
- NICHD NIH HHS [R03 HD046553-02, R03 HD046553-02S1, R03 HD046553-03, R03 HD046553-01A1] Funding Source: Medline
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Oocytes, in general, are greatly enriched in mitochondria to support higher rates of macromolecular synthesis and critical physiological processes characteristic of early development. An inability of these organelles to amplify and/or to accumulate ATP has been linked to developmental abnormality or arrest. The number of mitochondrial genomes present in mature mouse and human metaphase II oocytes was estimated by fluorescent rapid cycle DNA amplification, which is a highly sensitive technique ideally suited to quantitative mitochondrial DNA (mtDNA) analysis in individual cells. A considerable degree of variability was observed between individual samples. An overall average of 1.59 x 10(5) and 3.14 x 10(5) mtDNA molecules were detected per mouse and human oocyte, respectively. Furthermore, the mtDNA copy number was examined in polar bodies and contrasted with the concentration in their corresponding oocytes. In addition, the density of mtDNA in a cytoplasmic sample was estimated in an attempt to determine the approximate number of mitochondria transferred during clinical cytoplasmic donation procedures as well as to develop a clinical tool for the assessment and selection of oocytes during in vitro fertilisation procedures. However, no correlation was identified between the mtDNA concentration in either polar bodies or cytoplasmic samples and their corresponding oocyte.
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