Investigation on redox status and gene expression related to larval cryopreservation in the Pacific oyster Crassostrea gigas

Liu, Yibing; Zhan, Xin; Catalano, Sarah R.; Qin, Jianguang; Han, Jingfeng; Li, Xiaoxu

doi:10.1007/s12562-022-01594-1

Cited by 4 publications

(4 citation statements)

References 44 publications

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“…Three replicates were assessed for the control and treatment groups. The methods used by Liu et al [11,14] for larval culture, D-larvae rate (%) calculation, spat production, sample collection/assessment, survival rate (%) and relative survival rate (%) calculations at various developmental stages were followed.…”

Section: Larvae Preparationmentioning

confidence: 99%

“…cDNA amplification was carried out over 40 cycles under the following conditions: denaturation for 15 s at 95 • C, annealing for 1 min at 60 • C and a final melting gradient up to 95 • C using a ramp of 0.4 • C/s to check primer specificity. The relative abundance of genes was calculated using the 2 −∆Ct method [14].…”

Section: Quantification Of Mrnamentioning

confidence: 99%

“…Parameters such as larval performance (particularly the survival rate), morphology and organogenesis have been used to develop or improve the larval cryopreservation technique for marine bivalve species, but few studies have investigated the potential cryodamage that occurs at the molecular level [12][13][14]. Recently, studies on epigenetics against cryopreservation have attracted attention, with epigenetics being used as a tool to understand the mechanism of cryodamage [15,16].…”

Section: Introductionmentioning

confidence: 99%

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The Effects of Larval Cryopreservation on the Epigenetics of the Pacific Oyster Crassostrea gigas

Liu,

Bao,

Catalano

et al. 2023

IJMS

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High mortalities and highly variable results during the subsequent development of post-thaw larvae have been widely considered as key issues restricting the application of cryopreservation techniques to support genetic improvement programs and hatchery production in farmed marine bivalve species. To date, few studies have been undertaken to investigate the effects of cryodamage at the molecular level in bivalves. This study is the first to evaluate the effect of larval cryopreservation on the epigenetics of the resultant progenies of the Pacific oyster Crassostrea gigas. The results show that the level of DNA methylation was significantly (p < 0.05) higher and lower than that of the control when the trochophore larvae were revived and when they developed to D-stage larvae (day 1 post-fertilization), respectively, but the level returned to the control level from day 8 post-fertilization onwards. The expression of the epigenetic regulator genes DNMT3b, MeCP2, JmjCA, KDM2 and OSA changed significantly (p < 0.05) when the trochophore larvae were thawed, and then they reverted to the control levels at the D- and later larval developmental stages. However, the expression of other epigenetic regulator genes, namely, MBD2, DNMT1, CXXC1 and JmjD6, did not change at any post-thaw larval developmental stage. For the newly thawed trochophore larvae, the amount of methylated H3K4Me1 and H3K27Me1 significantly changed, and the expression of all Jumonji orthologs, except that of Jumonji5, significantly (p < 0.05) decreased. These epigenetic results agree with the data collected on larval performances (e.g., survival rate), suggesting that the effect period of the published cryopreservation technique on post-thaw larvae is short in C. gigas.

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Section: Larvae Preparationmentioning

confidence: 99%

Section: Quantification Of Mrnamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Effects of Larval Cryopreservation on the Epigenetics of the Pacific Oyster Crassostrea gigas

Liu,

Bao,

Catalano

et al. 2023

IJMS

Self Cite

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“…The application of transcriptome analysis technologies such as next-generation sequencing (NGS) is becoming widespread, using tools such as RNA sequencing (RNA-seq) that allow gene identification and their respective expression [ 16 , 17 ]. Gene expression have been successfully used in the investigation of sperm and larvae cryodamage in several species including bivalves [ 18 – 21 ]. There are studies in the use of RNA-seq to identify gene alterations in post-thaw sperm and embryos of different species [ 22 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

Comparative transcriptome analysis reveals molecular damage associated with cryopreservation in Crassostrea angulata D-larvae rather than to cryoprotectant exposure

Anjos,

Duarte,

Fatsini

et al. 2024

BMC Genomics

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Background 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. Results Expression analysis revealed 3 differentially expressed genes between larvae exposed to cryoprotectant solution and fresh larvae and 611 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. Conclusions 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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Effects of phosphatidylcholine and tocopherol during larval cryopreservation of Pacific oysters (Magallana gigas)

et al. 2023

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Investigation on redox status and gene expression related to larval cryopreservation in the Pacific oyster Crassostrea gigas

Cited by 4 publications

References 44 publications

The Effects of Larval Cryopreservation on the Epigenetics of the Pacific Oyster Crassostrea gigas

The Effects of Larval Cryopreservation on the Epigenetics of the Pacific Oyster Crassostrea gigas

Comparative transcriptome analysis reveals molecular damage associated with cryopreservation in Crassostrea angulata D-larvae rather than to cryoprotectant exposure

Effects of phosphatidylcholine and tocopherol during larval cryopreservation of Pacific oysters (Magallana gigas)

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