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Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish
Published on: April 22, 2017
Expression of microRNA processing machinery genes in rhesus monkey oocytes and embryos of different developmental
Namdori R Mtango1, Santhi Potireddy, Keith E Latham
1The Fels Institute for Cancer Research and Molecular Biology, Temple University Medical School, Philadelphia, Pennsylvania 19140, USA.
Abstract:
MicroRNAs (miRNAs) are a class of small RNAs that silence gene expression. In animal cells, miRNAs bind to the 3' untranslated regions of specific mRNAs and inhibit their translation. The correct regulation of mRNA expression by miRNAs is believed to be important for oocyte maturation, early development and implantation. We examined the expression of 25 mRNAs involved in the microRNA processing pathway in a nonhuman primate oocyte and embryo model. We observed that mRNAs related to miRNA splicing are downregulated during oocyte maturation while those related to miRNA processing are upregulated, indicating that there may exist a temporal difference in their activities related to transcriptional activity in germinal vesicle stage oocytes. We also observed that the vast majority of mRNAs examined were insensitive to alpha-amanitin at the 8-16 cell stage. The expression data did not reveal a major impact of embryo culture, and hormonal stimulation protocol affected only a small number of mRNAs, suggesting that the components of the pathway may be accumulated in the oocyte during oogenesis and resistant to exogenous insults. In comparison to published mouse array data, we observed species differences and similarities in the temporal expression patterns of some genes, suggesting that miRNA processing may be regulated differently. These data extend our understanding of the potential roles of miRNA during primate embryogenesis.
Insights
MicroRNA (miRNA) processing pathways are crucial for primate oocyte maturation and early development. Gene expression analysis reveals distinct temporal patterns and species-specific regulation, impacting embryogenesis.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are small RNAs that regulate gene expression by inhibiting mRNA translation.
- Proper miRNA regulation is vital for oocyte maturation, early development, and implantation.
- Understanding miRNA pathway dynamics is key to reproductive success.
Purpose of the Study:
- To investigate the expression patterns of 25 mRNAs in the miRNA processing pathway during nonhuman primate oocyte and embryo development.
- To identify temporal changes in miRNA pathway gene expression related to oocyte maturation and early embryogenesis.
- To compare primate miRNA processing gene expression with existing mouse data.
Main Methods:
- Analysis of mRNA expression levels for 25 genes involved in the miRNA processing pathway.
- Utilized a nonhuman primate oocyte and embryo model.
- Compared expression data with and without alpha-amanitin treatment and assessed effects of hormonal stimulation and embryo culture.
Main Results:
- mRNAs for miRNA splicing were downregulated, while those for miRNA processing were upregulated during oocyte maturation.
- Most examined mRNAs were insensitive to alpha-amanitin in the 8-16 cell stage embryos.
- Embryo culture had minimal impact, and hormonal stimulation affected few mRNAs, suggesting pathway component accumulation during oogenesis.
Conclusions:
- Primate oocyte maturation involves dynamic temporal shifts in miRNA pathway gene expression.
- The miRNA processing pathway appears robust and potentially pre-accumulated during oogenesis in primates.
- Species-specific differences exist in miRNA processing regulation compared to mice, highlighting unique primate developmental strategies.
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