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| Status |
Public on Jan 11, 2022 |
| Title |
The impact of ageing on the transition of the euploid human GV oocytes to in vivo matured MII oocytes |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background: Early embryonic development is governed by maternal transcripts stored within the oocyte during oogenesis. Transcriptional activity of the oocyte ultimately dictates its developmental potential and may be influenced by maternal age, resulting in reduced competence of oocytes derived from women of advanced age, compared with the young. In the current study, RNA-Seq was used to perform transcriptome profiling of human GV and MII oocytes derived from young and advanced maternal age women.
Participants/Materials and Methods: Cumulus dissection from donated oocytes was performed. GV and MII oocytes underwent deep RNA sequencing using the SMART-Seq v4 Ultra Low Input RNA protocol (Takara-Clontech, USA) and Nextera XT DNA library preparation kit (Illumina, USA). Data processing, quality assessment and bioinformatics analysis were performed using source-software, including FastQC, HISAT2, StringTie, edgeR and DAVID.
Results: Following deep single-cell RNA-Seq on GV and MII oocytes, hundreds of transcripts were significantly differentially expressed between young maternal age (YMA) and advanced maternal age (AMA) groups, with the most significant biological processes relating to mitochondrial reserves. The GV to MII transition shares common biological processes between young and AMA groups, however, some genes involved in mitochondria function were altered during ageing. A decrease in mitochondrial-related transcripts was also observed during the GV to MII transition. However, there was a much greater reduction of mitochondrial-related transcripts in MII oocytes of AMA. This observation was confirmed when YMA MII oocytes were compared with the AMA MII group with mitochondrial-related transcripts being significantly higher expressed in the YMA group, including biological processes, such as mitochondrial electron transport and ATP biosynthetic process. These results indicate a higher energy potential in YMA MII oocytes that is decreased with age. Other significantly higher biological processes in the YMA MII group include transcripts involved in the regulation of ubiquitin-dependent degradation. Lack of these transcripts could lead to a non-appropriate removal of oogenesis remnants following fertilisation in the AMA MII group.
Discussion: Understanding reproductive ageing effects at the RNA level in human oocytes may reveal differences in the mechanisms regulating the GV to MII transition that impact on oocyte quality in YMA and AMA patients. Further investigations of the up-/down-regulated transcripts during ageing could guide and improved IVF outcomes for AMA patients.
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| Overall design |
Fifteen patients treated for infertility in a single IVF unit agreed to participate in this study, where 5 GV and 5 MII oocytes derived from 8, 21-26 yo women (YMA group), and 5 GV and 6 MII oocytes from 7, 41-44yo women (AMA group). These oocytes were donated by women undergoing IVF treatment for research purposes. The samples were collected within a period of 3 months. RNA was isolated and deep sequenced at the single cell level.
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| Contributor(s) |
Ntostis P, Iles D, Kokkali G, Vaxevanoglou T, Kanavakis E, Pantou A, Huntriss J, Pantos K, Picton H |
| Citation(s) |
34755188 |
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| Submission date |
Jan 07, 2021 |
| Last update date |
Jan 11, 2022 |
| Contact name |
Panagiotis Ntostis |
| Organization name |
University of Leeds
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| Department |
Medicine and Health
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| Lab |
LICAMM
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| Street address |
Clarendon Way
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| City |
Leeds |
| ZIP/Postal code |
LS2 9JT |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
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| Samples (21)
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| Relations |
| BioProject |
PRJNA690226 |
| SRA |
SRP300738 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE164371_GEO_reads2sym_counts_submission.xlsx |
6.1 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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