Comparative transcriptome analysis reveals molecular damage associated with cryopreservation in Crassostrea angulata D-larvae rather than to cryoprotectant exposure
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ABSTRACT:
PROVIDER: PRJNA1035107 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR26660967_1.fastq.gz | Fastqsanger.gz | |||
| SRR26660967_2.fastq.gz | Fastqsanger.gz | |||
| SRR26660968_1.fastq.gz | Fastqsanger.gz | |||
| SRR26660968_2.fastq.gz | Fastqsanger.gz | |||
| SRR26660969_1.fastq.gz | Fastqsanger.gz |
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Project description:The Portuguese oyster Crassostrea angulata, a bivalve of significant economic and ecological importance, has faced a decline in both production and natural populations due to pathologies, climate change, and anthropogenic factors. To safeguard its genetic diversity and improve reproductive management, cryopreservation emerges as a valuable strategy. However, the cryopreservation methodologies lead to some damage in structures and functions of the cells and tissues that can affect post-thaw quality. Transcriptomics may help to understand the molecular consequences related to cryopreservation steps and therefore to identify different freezability biomarkers. This study investigates the molecular damage induced by cryopreservation in C. angulata D-larvae, focusing on two critical steps: exposure to cryoprotectant solution and the freezing/thawing process. Expression analysis revealed 3 differentially expressed genes between larvae exposed to cryoprotectant solution and fresh larvae and 511 differentially expressed genes in cryopreserved larvae against fresh larvae. The most significantly enriched gene ontology terms were “carbohydrate metabolic process”, “integral component of membrane” and “chitin binding” for biological processes, cellular components and molecular functions, respectively. Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified the “neuroactive ligand receptor interaction”, “endocytosis” and “spliceosome” as the most enriched pathways. RNA sequencing results were validate by quantitative RT-PCR, once both techniques presented the same gene expression tendency and a group of 11 genes were considered important molecular biomarkers to be used in further studies for the evaluation of cryodamage. The current work provided valuable insights into the molecular repercussions of cryopreservation on D-larvae of Crassostrea angulata, revealing that the freezing process had a more pronounced impact on larval quality compared to any potential cryoprotectant-induced toxicity. Additionally, was identify 11 genes serving as biomarkers of freezability for D-larvae quality assessment. This research contributes to the development of more effective cryopreservation protocols and detection methods for cryodamage in this species.
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Project description:Deep sequencing of mRNA from Pacific oyster Crassostrea gigas Competent larvae of Crassostrea gigas were treated with epinephrine solution, and then sampled at different time intervals. For shell damage experiment, shell were broken and then tissues were sampled at different time intervals.
2014-07-26 | E-GEOD-31011 | biostudies-arrayexpress
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Project description:The aim of this study was to gain insight into the molecular mechanisms of intraspecies difference of copper accumulation in Crassostrea angulata. In this attempt, we have performed a comprehensive analysis of the transcriptome of oyster (gill and mantle). Digital gene expression (DGE) technology was applied to analyze the relationships between gene expression and differential Cu body burden.
2016-06-30 | GSE65653 | GEO
Project description:The aim of this study was to gain insight into the molecular mechanisms of intraspecies difference of copper accumulation in Crassostrea angulata. In this attempt, we have performed a comprehensive analysis of the transcriptome of oyster (gill and mantle). Digital gene expression (DGE) technology was applied to analyze the relationships between gene expression and differential Cu body burden. Digital gene expression (DGE) technology was applied to analyze the relationships between gene expression and differential Cu body burden
2016-06-30 | E-GEOD-65653 | biostudies-arrayexpress
Project description:Increasing atmospheric CO2 raises sea surface temperatures and results in ocean acidification, which will impact upon calcifying marine organisms, such as the commercially and ecologically important Pacific oyster (Crassostrea gigas). Larval stages of development are particularly sensitive to such stressors and may represent population bottlenecks. A two-dimensional electrophoresis (2-DE) proteomic approach was used to investigate the response of 40 hour C. gigas larvae to ocean acidification and warming, and to relate protein expression to phenotypic variation in size and calcification. Larvae were reared at two pHs (8.1 and 7.9) and two temperatures (20°C and 22°C), and comparisons carried out between the four possible treatment combinations. In total 22 differentially expressed spots, corresponding to 18 proteins, were identified by nano-liquid chromatography tandem mass spectrometry. These proteins had roles in metabolism, biomineralisation, intra- and extra-cellular matrix formation and as molecular chaperones. Thirteen of these spots responded to acidification, of which 11 showed reduced expression during acidification. Declines in ATP synthase, arginine kinase and other metabolic proteins suggest metabolic depression occurred during acidification and reduced protein synthesis. In contrast, 6 of 7 proteins that were differentially expressed during warming showed increased expression. Among these were molecular chaperones including protein disulphide isomerase (PDI) and Grp78. Concurrent acidification and warming appeared to mitigate some proteomic changes and negative phenotypic effects observed in acidification at 20°C; however, differential expression of nine proteins and other temperature-independent effects on calcification phenotypes suggest that larval responses to multiple stressors will be complex.
2022-03-01 | PXD002316 | Pride
Project description:Cryopreservation is an important technique for preserving fertility potential through ovarian tissue banking. Slow freezing has traditionally been the standard protocol used for ovarian tissue cryopreservation. However, vitrification, as an emerging method, is considered to be an alternative to slow freezing, thus generating some debate in the field. This study utilizes high-resolution spatial transcriptomics to comprehensively profile and compare the molecular impacts of two cryopreservation methods - slow freezing and vitrification - on human ovarian tissues at an unprecedented resolution. Over 135,000 spots were profiled from Fresh and cryopreserved ovarian tissue slices, identifying 8 major cell types and 25 subtypes through marker expression. Detailed spatial localization patterns revealed heterogeneous stromal cell subpopulations and ovarian structures. Transcriptional profiling showed that both cryopreservation methods significantly repressed gene expression, indicative of tissue damage. However, slow freezing induced a broader pro-survival inflammatory and tissue remodeling response compared to vitrification. Gene set enrichment analysis identified elevated ribosomal processes, inflammatory signaling pathways like TNF-?/NF-?B and IL-6/STAT3, as well as extracellular matrix (ECM) organization and collagen synthesis as dominant response themes in slow frozen tissue, which were observed to a lesser degree in vitrified tissue. Key response drivers included MYC, TIMP1, and DCN, with stromal cells located in the cortical regions showing the strongest pathway enrichment. Ribosomal gene expression displayed spatial gradients and supported stromal responses after freezing. We finally demonstrated signaling pathway differences between the two cryopreservation methods, noting TGF-? as a highly possible pathway responsible for coordinating with certain endothelial subgroups to enhance ECM deposition. In summary, this study provides a comprehensive atlas of cell types and their spatial structures in human ovary. It reveals that slow freezing elicits a strong but balanced inflammatory and tissue regenerative state compared to the vitrification. It also offers insights on cryoinjury to ovarian stroma, which may guide optimization of ovarian tissue cryopreservation protocols for clinical applications.
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