Dataset Information

Small Noncoding RNA Signatures for Determining the Developmental Potential of an Embryo at the Morula Stage.

ABSTRACT: As part of the optimization of assisted reproductive technology programs, the aim of the study was to identify key small noncoding RNA (sncRNA) molecules that participate in maternal-to-zygotic transition and determine development potential and competence to form a healthy fetus. Small RNA deep sequencing followed by quantitative real-time RT-PCR was used to profile sncRNAs in 50 samples of spent culture medium from morula with different development potentials (no potential (degradation/developmental arrest), low potential (poor-quality blastocyst), and high potential (good/excellent quality blastocyst capable of implanting and leading to live birth)) obtained from 27 subfertile couples who underwent in vitro fertilization. We have shown that the quality of embryos at the morula stage is determined by secretion/uptake rates of certain sets of piRNAs and miRNAs, namely hsa_piR_011291, hsa_piR_019122, hsa_piR_001311, hsa_piR_015026, hsa_piR_015462, hsa_piR_016735, hsa_piR_019675, hsa_piR_020381, hsa_piR_020485, hsa_piR_004880, hsa_piR_000807, hsa-let-7b-5p, and hsa-let-7i-5p. Predicted gene targets of these sncRNAs included those globally decreased at the 8-cell-morula-blastocyst stage and critical to early embryo development. We show new original data on sncRNA profiling in spent culture medium from morula with different development potential. Our findings provide a view of a more complex network that controls human embryogenesis at the pre-implantation stage. Further research is required using reporter analysis to experimentally confirm interactions between identified sncRNA/gene target pairs.

SUBMITTER: Timofeeva A

PROVIDER: S-EPMC7764539 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Small Noncoding RNA Signatures for Determining the Developmental Potential of an Embryo at the Morula Stage.

Timofeeva Angelika A Drapkina Yulia Y Fedorov Ivan I Chagovets Vitaliy V Makarova Nataliya N Shamina Maria M Kalinina Elena E Sukhikh Gennady G

International journal of molecular sciences 20201210 24

As part of the optimization of assisted reproductive technology programs, the aim of the study was to identify key small noncoding RNA (sncRNA) molecules that participate in maternal-to-zygotic transition and determine development potential and competence to form a healthy fetus. Small RNA deep sequencing followed by quantitative real-time RT-PCR was used to profile sncRNAs in 50 samples of spent culture medium from morula with different development potentials (no potential (degradation/developm ...[more]

PMID: 33321810

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Project description:For in vitro produced embryos generated from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) procedure, the intra- and extra-environmental factors during in vitro culture have significant impact on latter embryo development and fetus growth. Assisted hatching (AH), an effective approach to facilitate hatchability for in vitro generated embryos, is an essential step for successful embryo implantation in the uterus. However, regarding the different AH methods reported in clinical practice, it is still unknown whether zona pellucida (ZP) broken is based on AH applied in diverse stages of embryos affect implantation and fetal development. Here, piezo-mediated AH treatments were classified into four categories: (1) drilling one small hole (SH) with a diameter of 10 μm on ZP (SH); (2) drilling one large hole (LH) with a diameter of 40 μm on ZP (LH); (3) made a small area with diameter of 40-μm thinner on ZP [small area thinner (ST)]; (4) made a large area with a diameter of 80-μm thinner [large area thinner (LT)]. These four AH treatments were applied in different stage embryos including two-cell, four-cell, and morula. The most efficient AH approach was chosen according to the final hatch rate at 120 h after fertilization. We found that the approach of SH applied in morula-stage embryos obtained the highest hatch rate. To further investigate if this treatment has any side effect on later development after embryo transfer, we evaluated embryo implantation, gestational period, litter size, and growth. Our results showed that SH applied in morula-stage embryos could facilitate the implantation process and increase litter size. Meanwhile, this approach had no side effect on birth weight, growth, or gender ratio in the offspring. We conclude that drilling a SH on ZP in morula-stage embryos is an effective and reliable AH approach for in vitro cultured embryos in rodent. And this approach is worth further investigating in human-assisted reproductive technology.

Project description:BACKGROUND: While initially sensitive to heat shock, the bovine embryo gains thermal resistance as it progresses through development so that physiological heat shock has little effect on development to the blastocyst stage by Day 5 after insemination. Here, experiments using 3' tag digital gene expression (3'DGE) and real-time PCR were conducted to determine changes in the transcriptome of morula-stage bovine embryos in response to heat shock (40 degrees C for 8 h) that could be associated with thermotolerance. RESULTS: Using 3'DGE, expression of 173 genes were modified by heat shock, with 94 genes upregulated by heat shock and 79 genes downregulated by heat shock. A total of 38 differentially-regulated genes were associated with the ubiquitin protein, UBC. Heat shock increased expression of one heat shock protein gene, HSPB11, and one heat shock protein binding protein, HSPBP1, tended to increase expression of HSPA1A and HSPB1, but did not affect expression of 64 other genes encoding heat shock proteins, heat shock transcription factors or proteins interacting with heat shock proteins. Moreover, heat shock increased expression of five genes associated with oxidative stress (AKR7A2, CBR1, GGH, GSTA4, and MAP2K5), decreased expression of HIF3A, but did not affect expression of 42 other genes related to free radical metabolism. Heat shock also had little effect on genes involved in embryonic development. Effects of heat shock for 2, 4 and 8 h on selected heat shock protein and antioxidant genes were also evaluated by real-time PCR. Heat shock increased steady-state amounts of mRNA for HSPA1A (P<0.05) and tended to increase expression of HSP90AA1 (P<0.07) but had no effect on expression of SOD1 or CAT. CONCLUSIONS: Changes in the transcriptome of the heat-shocked bovine morula indicate that the embryo is largely resistant to effects of heat shock. As a result, transcription of genes involved in thermal protection is muted and there is little disruption of gene networks involved in embryonic development. It is likely that the increased resistance of morula-stage embryos to heat shock as compared to embryos at earlier stages of development is due in part to developmental acquisition of mechanisms to prevent accumulation of denatured proteins and free radical damage.

Dataset Information

Small Noncoding RNA Signatures for Determining the Developmental Potential of an Embryo at the Morula Stage.

Publications

Small Noncoding RNA Signatures for Determining the Developmental Potential of an Embryo at the Morula Stage.

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