Imaging Mitochondrial Organization in Living Primate Oocytes and Embryos using Multiphoton Microscopy

We employed multiphoton laser scanning microscopy (MPLSM) to image changes in mitochondrial distribution in living rhesus monkey embryos. This method of imaging does not impair development; thus, the same specimen can be visualized multiple times at various developmental stages. Not only does this i...

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Bibliographic Details
Published in:Microscopy and microanalysis 2003-06, Vol.9 (3), p.190-201
Main Authors: Squirrell, J.M., Schramm, R.D., Paprocki, A.M., Wokosin, D.L., Bavister, B.D.
Format: Article
Language:English
Subjects:
Animals
Cytoplasm - metabolism
Developmental stages
Diagnostic Imaging - methods
Embryo, Mammalian - metabolism
Embryo, Mammalian - ultrastructure
Embryonic and Fetal Development
Embryos
Female
Macaca mulatta - embryology
Male
Microscopy, Fluorescence, Multiphoton
Mitochondria - physiology
Mitochondria - ultrastructure
Oocytes - growth & development
Oocytes - ultrastructure
Staining and Labeling
Time Factors
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Summary:We employed multiphoton laser scanning microscopy (MPLSM) to image changes in mitochondrial distribution in living rhesus monkey embryos. This method of imaging does not impair development; thus, the same specimen can be visualized multiple times at various developmental stages. Not only does this increase the amount of information that can be gathered on a single specimen but it permits the correlation of early events with subsequent development in the same specimen. Here we demonstrate the utility of MPLSM for determining changes in mitochondrial organization at various developmental stages and show that rhesus zygotes possess a distinct accumulation of mitochondria between the pronuclei prior to syngamy. We present evidence that suggests that this pronuclear accumulation may be positively correlated with development to the blastocyst stage—in the same embryo—thereby illustrating how MPLSM can be used to correlate cellular dynamics of primate oocytes and early embryos with their developmental potential. Understanding the relationship between mitochondrial distribution and the subsequent development of mammalian embryos, particularly primates, will increase our ability to improve embryo culture technologies, including those used for human assisted reproduction.
ISSN:1431-9276
1435-8115
DOI:10.1017/S1431927603030174