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Insights into the digestive gland of Mytilus coruscus under heat stress using TMT-based proteomics

ABSTRACT: A healthy existence relationship between mussels and environment relies on suitable PH, adequate oxygen and suitable temperature. Besides ocean acidification, ocean warming has been shown to cause mussel dead, a phenomenon responsible for extensive mussel dead around the word. How heat stress impacts mussel alive is poorly understood. We founded that. The superoxide dismutase (SOD) activity of two treatment groups were significantly higher than that of control group, and the activity of acid phosphatase (ACP) was significantly higher in two treatment groups than control group. The lactate dehydrogenase (LDH) activity of 26 ℃ treatment group was significantly higher than that of control group, and the LDH activity of 33 ℃ treatment group was significantly lower than that of control group. Cellular content of lysozyme (LZM) was significantly higher in two treatment groups than control group. In this study, we applied TMT quantification to investigate how Mytilus coruscus responds to heat stress (26 ℃, 33 ℃). We identified 1652 to 1878 differentially expressed proteins (DEPs) from these heat conditions, among which 897 were commonly regulated by all the two temperature and were significantly enriched in nutrients digestion and absorption (e. g. fat digestion and absorption). Furthermore, proteins associated with nutrients metabolism, signaling pathways (p53 signaling pathway and MAPK signaling pathway)and energy metabolism (e. g. TCA cycle) were impacted, suggesting that the ability of basic metabolism of Mytilus coruscus may be altered by heat stress.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mytilus Coruscus

TISSUE(S): Digestive Gland

SUBMITTER: 乐钟许

LAB HEAD: Insights into the digestive gland of Mytilus coruscus under heat stress using TMT-based proteomics

PROVIDER: PXD035618 | Pride | 2023-05-30

REPOSITORIES: Pride

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Project description:Direct comparison of the transcriptional patterns between male and female in the digestive gland of a natural population of the marine mussel Mytilus galloprovincialis sampled in the Bizerta Lagoon, Tunisia, across November 2007 -February March 2008 (four stages, winter peak). Background: Seasonal environmental changes may affect the physiology of Mytilus galloprovincialis (Lam.), an intertidal filter-feeder bivalve occurring commonly in Mediterranean and Atlantic coastal areas. We investigated seasonal variations in relative transcript abundance of the digestive gland and the Digestive gland (gonads) of males and females. To identify gene expression trends, we used a medium-density cDNA microarray (1.7 K probes) in dual-color competitive hybridization analyses. Results: Hierarchical clustering of digestive gland microarray data showed two main branches, distinguishing profiles associated with the “hot” months (May–August) from the other months. Genes involved in chitin metabolism, associated with mussel nutrition and digestion, showed higher expression during summer. Moreover, we found different gene expression patterns in the digestive glands of males and females during the four stages of mussel gonadal development. Microarray data from gonadal transcripts also displayed clear patterns during the different developmental phases with peak relative mRNA abundance at the ripe phase (stage III) for both sexes. Conclusion: These data showed a clear temporal pattern in gene expression profiles of mussels sampled over an annual cycle. Physiological response to thermal variation, food availability, and reproductive status across months may contribute to variation in gene expression. Test/reference design (female/male). Direct comparison of RNA extracts obtained from the Digestive gland tissue of female and male animals. Two (male, female) x four conditions (gonad developmental stage 1, stage 2, stage 3, stage 4). Dual color competitive hybridizations with label swap. Single individuals. Four biological replicates. One replicate per array.

Project description:Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to copper along with a temperature gradient Background: Global warming is a major factor that may affect biological organization, especially in marine ecosystems and in coastal areas that are particularly subject to anthropogenic pollution. We evaluated the effects of simultaneous changes in temperature and copper concentrations on lysosomal membrane stability of the blue mussel Mytilus galloprovincialis (Lam.). Temperature and copper exerted additive effects on lysosomal membrane stability, exacerbating the toxic effects of metal cations present in non-physiological concentrations. Mussel lysosomal membrane stability was positively related toscope for growth, indicating possible effects of increasing temperature on mussel populations in metal-polluted areas. To clarify the molecular response to environmental stressors, we used a cDNA microarray with 1,700 sequences to measure the relative transcript abundances in the gills of mussels exposed to copper (40M-BM-5g/L) and a temperature gradient (16M-BM-0C, 20M-BM-0C, and 24M-BM-0C). In animals exposed only to heat stress, hierarchical clustering of the microarray data revealed three main clusters, which were largely dominated by down-regulation of translation-related differentially expressed genes, drastic up-regulation of folding protein-related genes, and genes involved in chitin metabolism. The response of mussels exposed to copper at 24M-BM-0 C was characterized by an opposite pattern of the genes involved in translation, most of which were up-regulated, as well asthe down-regulation of genes encoding heat shock proteins and microtubule-based movement proteins. Our data provide novel information on the transcriptomic modulations in mussels facing temperature increases and high copper concentrations; these data highlight the risk of marine life exposed to toxic chemicals in the presence of temperature increases due to climate change. Digestive gland tissue from individual animals in different experimental conditions were analyzed in a complete loop design. Dual color competitive hybridizations (Control M-bM-^@M-^\16M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^] vsM-bM-^@M-^]20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]; M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]) including label swap. Single individuals. Four biological replicates. One replicate per array.

Project description:Transcriptional profiling of natural population of mussels (Mytilus galloprovincialis) -digestive gland tissue- comparing female individuals sampled in the Bizerta Lagoon, Tunisia, across May 2007 - April 2008. Background: Seasonal environmental changes may affect the physiology of Mytilus galloprovincialis (Lam.), an intertidal filter-feeder bivalve occurring commonly in Mediterranean and Atlantic coastal areas. We investigated seasonal variations in relative transcript abundance of the digestive gland and the mantle (gonads) of males and females. To identify gene expression trends, we used a medium-density cDNA microarray (1.7 K probes) in dual-color competitive hybridization analyses. Results: Hierarchical clustering of digestive gland microarray data showed two main branches, distinguishing profiles associated with the M-bM-^@M-^\hotM-bM-^@M-^] months (MayM-bM-^@M-^SAugust) from the other months. Genes involved in chitin metabolism, associated with mussel nutrition and digestion, showed higher expression during summer. Moreover, we found different gene expression patterns in the digestive glands of males and females during the four stages of mussel gonadal development. Microarray data from gonadal transcripts also displayed clear patterns during the different developmental phases with peak relative mRNA abundance at the ripe phase (stage III) for both sexes. Conclusion: These data showed a clear temporal pattern in gene expression profiles of mussels sampled over an annual cycle. Physiological response to thermal variation, food availability, and reproductive status across months may contribute to variation in gene expression. Loop design: Female individuals from 12 different months across one year (May 2007 to April 2008). Dual color competitive hybridizations (month n vs month n+1) including dye swap. Single individuals. Three (May 2007 to December 2007) or four biological replicates (January 2008 to April 2008 ). One replicate per array.

Project description:Transcriptional profiling of mussel (Mytilus galloprovincialis) digestive gland tissue comparing control tissue with tissue obtained from animals exposed to sublethal amounts of Chlorpyrifos, animals injected into the posterior adductor muscle with 25 pico-moles 17β-estradiol (E2) and animals pre-exposed for three days to the pesticide and further injected with E2. Background: Many pesticides have been shown to act as endocrine disrupters. Although the potencies of currently used pesticides as hormone agonists/antagonists in vitro are low compared with those of natural ligands, their ability to act via multiple mechanisms might enhance the biological effect. The organophosphate Chlorpyrifos (CHP) has been shown to be weakly estrogenic and cause adverse neurodevelopmental effects in mammals. However, no information is available on the possible endocrine effects of CHP in aquatic organisms. In the digestive gland of the bivalve Mytilus galloprovincialis, a target tissue for the action of both estrogens and pesticides, the possible effects of CHP on the responses to the natural estrogen 17β-estradiol (E2) were investigated. Methodology/Principal findings: Mussels were exposed to CHP (4.5 mg/l, 72 hrs) and subsequently injected with E2 (6.75 ng/g dw). Responses were evaluated in CHP, E2 and CHP/E2 treatment groups at 24 h p.i. by a biomarker/transcriptomic approach. CHP and E2 induced additive, synergistic, and antagonistic effects on lysosomal biomarkers (lysosomal membrane stability, lysosome/cytoplasm volume ratio, lipofuscin and neutral lipid accumulation). Additive and synergistic effects were also observed on the expression of estrogen-responsive genes (GSTπ, catalase and 5-HTR) evaluated by RT-Q-PCR. The use of a 1.7K cDNA Mytilus microarray showed that CHP, E2 and CHP/E2, induced 81, 44, and 65 Differentially Expressed Genes (DEGs), respectively. 24 genes were exclusively shared between CHP and CHP/E2, only 2 genes between E2 and CHP/E2. Moreover, 36 genes were uniquely modulated by CHP/E2. Gene ontology annotation was used to elucidate the putative mechanisms involved in the responses elicited by different treatments. Conclusions: The results show complex interactions between CHP and E2 in mussel digestive gland, indicating that the combination of certain pesticides and hormones may give rise to unexpected effects at the molecular/cellular level. Overall, these data demonstrate that CHP can interfere with the mussel responses to natural estrogens. Four-condition experiment. Dual color competitive hybridizations. Common reference (vehicle treated animals, 0.02% Dimethyl sulfoxide, injected with 50 μl of a solution of Artificial Sea Water containing 0.05 % ethanol); Pools of six animals. Biological replicates: 4 controls, 4 Chlorpyrifos, 4 17β-estradiol, 4 Chlorpyrifos-17β-stradiol. One replicate per array.

Project description:Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to TCDD, n-TiO2 and their binary mixture Background: Exposure of marine organisms to pollutant mixtures may affect the pattern of contaminant uptake/bioaccumulation, as well as of gene expression in the tissues. Despite the growing concern over the potential biological impact of nanoparticles (NPs) in the aquatic environment, little is known about their interactions with other pollutants.We have recently shown that in the marine mussel Mytilus galloprovincialis exposure to n-TiO2, one of the most widespread type of NPs in use, in combination with 2,3,7,8-TCDD, chosen as model organic xenobiotic, can exert antagonistic or synergistic effects on different biomarkers from the molecular to the tissue level, depending on cell/tissue and type of measured response. An integrated approach involving immunhistochemical and transcriptomic analysis was employed to clarify the itteractive effects of n-TiO2 and TCDD in mussels digestive gland. In particular,TCDD bioaccumulation was evaluated utilizing specific anti-TCDD fluorescent antibodies. Moreover, immunohistochemical evaluation of antioxidant and cytoskeletal components was performed. To provide clues about how the molecular response to the investigated compounds is modulated, we used a cDNA microarray with1673 sequences. In animals exposed only to TiO2, functional genomics analysis of the microarray data (48 differentially expressed genes (DEGs)) highlighted three biological processes, largely dominated by the up-regulation of microtubule-based movement-related genes. Exposure to 2,3,7,8-TCDD yielded 49 DEGs exhibiting distinct patterns in terms of biological processes. Finally, exposure to the mixture rendered 62 GEGs characterized by the regulation of response to chemical stimulus, microtubule-based movement and intracellular signal transduction. Our data should be carefully considered in view of the biological effects of emerging pollutants, particularly in case of mixture chemicals. Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to TCDD, n-TiO2 and their binary mixture

Project description:Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to nickel along with a temperature gradient Background: The exposure of marine organisms to stressing agents may affect the level and pattern of gene expression. Although many studies have examined the ecological effects of heat stress on mussels, little is known about the physiological mechanisms that might be affected by co-exposure to heat stress and environmental contaminants such as nickel (Ni). In the present work we investigated the effects of simultaneous changes in temperature and Ni supply on lysosomal membrane stability (LMS) and malondialdehyde accumulation (MDA) in the digestive gland (DG) of the blue mussel Mytilus galloprovincialis (Lam.). To shed some light into how the molecular response to environmental stressors is modulated, we employed a cDNA microarray with 1,673 sequences to measure the relative transcript abundances in the DG of mussels exposed to Ni along with the temperature increase. Temperature and Ni rendered additive effects on LMS and MDA accumulation, increasing the toxic effects of metal cations. Ni loads in DG tissues was also affected by co-exposure to 26M-BM-0C. In animals exposed only to heat stress, functional genomics analysis of the microarray data (171 DEGs) revealed 7 biological processes, largely dominated by the up-regulation of folding protein-related genes, and the down regulation of genes involved in cell migration and cellular component assembly. Exposure to Ni at 18M-BM-0C and 26M-BM-0C rendered respectively 188 and 262 DEGs showing distinct pattern in term of biological processes. In particular, the response of mussels exposed to Ni at 26M-BM-0C was characterized by the up regulation of proteolysis, ribosome biogenesis, response to unfolded proteins and catabolic-related genes as well as the down-regulation of genes encoding cellular metabolic processes. Our data provide new insights on the transcriptomic response in mussels challenging temperature increases and Ni exposure and should be carefully considered in view of the biological effects of heat stress and particularly in polluted areas. Digestive gland tissue from individual animals in different experimental conditions were analyzed in a complete loop design. Dual color competitive hybridizations (Control M-bM-^@M-^\16M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^] vsM-bM-^@M-^]20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]; M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]) including label swap. Single individuals. Four biological replicates. One replicate per array.

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Insights into the digestive gland of Mytilus coruscus under heat stress using TMT-based proteomics

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