Gene expression profile of in vitro produced bovine blastocyst with and without Thyroid hormone supplementation
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ABSTRACT:
ORGANISM(S): Bos taurus
PROVIDER: GSE58901 | GEO | 2014/09/01
SECONDARY ACCESSION(S): PRJNA253832
REPOSITORIES: GEO
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Project description:Bovine IVP blastocysts (day 8) were produced with and without thyroid hormones and gene expression profiles were compared. The IVC media (SOF) was supplemented with 50ng/ml of each T3 and T4. Four independent IVF run, each gave 1 control and 1 treated sample. Each sample is a pool of 5 blastocyst. Total Rna was extracted from each sample, amplified, labeled (also with dye swap) and hybridized to the EmbryoGene custom Agilent array (GPL13226). FlexArray software was used for data analysis.
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Project description:Production of embryos in vitro has enormous potential for research and commercial applications. Unfortunately, in vitro production of porcine embryos is extremely inefficient. Despite the characterization of distinct phenotypes, little is known about the molecular mechanisms and altered physiological processes that account for poor IVP development. The objective of the current study was to compare global gene expression patterns from IVO and IVP embryos using small amplified RNA (SAR)-SAGE. Whole-cell RNA from pools of Day 6 in vivo-(IVO) and in vitro-produced (IVP) blastocysts was used to construct SAR-SAGE libraries. Sequence analysis of the IVO and IVP libraries yielded a total of 98,771 and 98,408 tags, respectively. A total of 20,029 and 23,453 unique putative transcripts were detected in the IVO and IVP libraries, respectively. Statistical analyses of SAGE tag frequencies between the IVO and IVP libraries indicated that 938 and 193 tags were differentially expressed at a P < 0.05 and P < 0.001 level of significance, respectively. Tentative annotation of the differentially expressed SAGE tags was determined using BLAST sequence alignment with the TIGR porcine specific gene index (SSGI) and cross-species alignment using RepeatMasker to determine homologous human orthologs. Annotated tags were categorized into functional groupings according to gene ontology annotations. Real-time PCR was used to confirm differential expression for several transcripts from IVO and IVP blastocysts. These results demonstrate compromised gene expression from IVP blastocysts compared with IVO blastocysts for a number of biological processes including cellular metabolism, organization and response to stress; thereby providing potential target pathways for improvement of IVP methods. Keywords: Comparative (in vivo- vs. in vitro-produced porcine embryos) Whole-cell RNA from pools of Day 6 in vivo- and in vitro-produced blastocysts was used to construct small amplified RNA (SAR)-SAGE libraries.
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Project description:There is an inevitable need for combining in vitro and in vivo culture systems during specific developmental time points to facilitate a comprehensive understanding of early embryo development which would yield insights into the molecular pathways controlling early development and to improve our knowledge in regulation of embryonic development. In current study, we aimed to understand the influences of alternative culture conditions (in vivo or in vitro) during EGA event on embryonic developmental rate, gene expression pattern and subsequent influences on pathways and biological functions controlling bovine embryo development. Six different blastocyst groups were produced under alternative in vivo and in vitro culture conditions. The first two groups (Vitro_4-cell and Vitro_16-cell) were matured, fertilized and cultured in vitro until 4- and 16-cell stage, respectively then transferred to synchronized recipients and continued culture in vivo until day 7 blastocyst stage. The second two groups (Vivo_4-cell and Vivo_16-cell) were matured, fertilized and cultured in vivo until 4- and 16-cell stage, respectively then flushed out and cultured in vitro until day 7 blastocyst stage. Complete in vitro (IVP) and in vivo (control group) blastocysts were also produced. Samples from the three pools (biological replicates) of each blastocyst group and in vivo control blastocyst were hybridized on EmbryoGENE’s bovine microarray using a dye-swap design (technical replicates) for a total of six arrays.
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Project description:We investigated the effects of thyroid hormone disruptions on gene expression in juvenile mice liver to develop a stronger understanding of the mechanisms by which thyroid disrupting chemicals impair development. Gene expression was examined by hybridization of hepatic RNA to Agilent mouse microarrays for hyper-, hypo-, hypo-replacement (hypo+) and euthyroid animals. Keywords: Toxicogenomics, biomarkers of thyroid disruptors Hypothyroidism was induced from post natal day (PND) 13 to 15 by adding model thyroid toxicants methimazole and sodium perchlorate to drinking water of pregnant females. Hyperthyroidism was induced by intraperitoneal injections (i.p.) of THs at PND 15, 4 hours before decapitation and tissue collection. For the hypothyroid/replacement group; dams were provided with drinking water for 3 days (PND 13 to 15), containing a mixture of methimazole/sodium perchlorate. Pups then received intraperitoneal injections of thyoid hormones on PND 15, 4 hours before decapitation and tissue collection.
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Project description:Thyroid hormones are important for homeostatic control of energy metabolism and body temperature. Although skeletal muscle is considered an important site for thyroid action, the contribution of thyroid hormone receptor signaling, in muscle, to whole-body energy metabolism and body temperature has not been resolved. Here, we show that thyroid hormone-induced increase in energy expenditure requires thyroid hormone receptor alpha 1 (TRa1) in skeletal muscle, but that thyroid hormone induced elevation in body temperature is independent of muscle-TRa1. In slow-twitch soleus muscle, ablation of TRa1 leads to an altered fiber type composition toward a more oxidative phenotype, which, however, does not influence running capacity or motivation to voluntary running. RNA-sequencing of soleus muscle from WT mice and TRaHSACre mice revealed differentiated transcriptional regulation of genes associated with muscle thermogenesis, such as sarcolipin and UCP3, thus providing molecular clues pertaining to the mechanistic underpinnings of TRa1-linked control of whole-body metabolic rate. Together, this work establishes a fundamental role for skeletal muscle in thyroid hormone-stimulated increase in whole-body energy expenditure.
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