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SRX22756893: GSM7937318: pool of embryo, low concentration of P4, rep1; Bos taurus; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 85.4M spots, 17.3G bases, 5Gb downloads

External Id: GSM7937318_r1
Submitted by: Sciences Animales, Université Laval
Study: Embryo response to different progesterone concentrations during superovulation of Holstein heifers
show Abstracthide Abstract
Greater progesterone (P4) concentrations during the follicular growth have been associated with greater quality of embryos produced by superovulated cows and heifers. However, it is yet to be determined the mechanisms by which oocyte exposure to greater P4 concentrations improves early embryo quality. The objective of this study was to evaluate the impact of different P4 concentrations during superovulation on the transcriptome profile of early bovine embryos. A total of 63 post-puberty Holstein heifers were randomly assigned into two experimental groups: High P4 (n = 31) and Low P4 (n = 32). Heifers received a pre-synchronization protocol followed by a protocol of superovulation that included the allocated P4 treatment. Embryo collection was performed 7 days post artificial insemination and embryos were evaluated for stage of embryonic development and grades of quality. Embryos graded as good/excellent quality (High P4: n = 27; Low P4: n = 27) were randomly allocated in 3 biological replicates per treatment group, balanced for stage of embryonic development. Transcriptome analysis suggested that exposure to different concentrations of P4 during superovulation promote changes in gene expression of 7 days old embryos that may be related to their developmental competence. These modifications are associated with downregulation of beta-estradiol pathway, upregulation of trophoblast-related genes and downregulation of WNT signalling pathway. Our results suggest a potential sensitivity of future embryos to follicular P4 levels but do not allow to conclude if the effect is from the oocyte, the oviduct, or the uterus response to P4. Overall design: Heifers underwent to a pre-synchronization protocol followed by a protocol of superovulation that included the allocated P4 treatment. Embryos were collected 7 d post-artificial insemination (AI) and were evaluated for fertilization, stage of embryonic development, and grades of quality. Embryos were randomly allocated in 3 biological replicates per treatment group on which we performed gene expression profiling using RNAseq. Finally, we identified differential expression of genes between treatment groups.
Sample: pool of embryo, low concentration of P4, rep1
SAMN38658937 • SRS19744443 • All experiments • All runs
Organism: Bos taurus
Library:
Name: GSM7937318
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Embryo total RNA was extracted from each biological replicate using the PicoPure RNA isolation kit (Thermo Fisher Scientific) with DNAse treatment (Qiagen) as per the manufacturer's protocol. RNA was eluted in 15 µL of nuclease-free water and kept at -80°C for subsequent library preparation. RNA concentration and integrity were evaluated using an Agilent 4200 TapeStation (Agilent Technologies) with the High Sensitivity RNA ScreenTape Assay (Agilent Technologies). RNA Integrity Number (RIN) ranged from 5.0 to 8.3 RIN with an average of 6.2 ± 1.4 RIN and RNA concentration ranged from 308 to 556 pg/µL with an average of 414 ± 113.6 pg/µL. Library preparation was performed for each biological replicate. Enrichment of mRNA was conducted using the NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490, New England Biolabs, USA) and libraries were prepared using the NEBNext Ultra II RNA Library kit for Illumina (E7770, E7775, New England Biolabs, USA). The library preparation protocol was followed as recommended by the manufacturer's guidelines for 10 ng – 1μg of DNA-free total RNA, with 15 cycles for the PCR enrichment of adaptor-ligated DNA and Illumina index primers (Illumina). Purified libraries were eluted in 23 μL of (0.1X) TE buffer and stored at -20°C for future analysis. The quality of the 6 libraries were verified using Agilent bioanalyzer (4200 TapeStation, Agilent Technologies) with the High Sensitivity D1000 ScreenTape Assay (Agilent Technologies) before libraries were pooled in equimolar proportion and sequenced with NovaSeq 6000 using paired-end 100bp protocol.
Runs: 1 run, 85.4M spots, 17.3G bases, 5Gb
Run# of Spots# of BasesSizePublished
SRR2706746385,409,74917.3G5Gb2024-06-04

ID:
30794115

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