Exopalaemon carinicauda in response to alkalinity stress
ABSTRACT: In this study, an integrative analysis of the gill-specific proteome at 0 h, 12 h and 36 h after alkalinity stress was performed to identify important regulators and pathways involved in alkalinity adaption of Exopalaemon carinicauda. This study reveals the first time-course, gill-specific, combined proteomic profiling associated with alkalinity adaption of E. carinicauda and provides new insights into the mechanisms underlying the molecular response to alkalinity stress in shrimp.
Project description:The experiment focused on the transcriptomic changes associated with gill inflammation in sea farmed Atlantic salmon (Salmo salar). To ensure the multifactorial aspect of gill inflammation, fish were sampled at three marine production sites (A on Isle of Mull, B in Shetland and C in Shetland) between October 2017 and March 2018. All fish were of strain Fanad and originated from the same egg fertilisation batch. They were reared in different hatcheries (Couldoran, Pettigo-Damph and Knock-Frisa for sites A, B and C, respectively) for one year and entered the sea in spring 2017. The resultant gill tissues (44 samples in total with 1 gill sample per fish) were first scored for proliferative gill disease (PGD), using gross morphology PGD scores from 0 with no visual pathology to 5 with severe visual pathology, and then subjected to RNA-seq and histopathological (microscopic) examination. One RNA-seq sample (fish 95) was identified as an outlier and removed from the subsequent analysis. As a result, the analysis aiming to integrate gill transcriptome, gross morphology and histopathology was performed on 43 gill samples, classified either as PGD score 1 (n = 26) or PGD score 3 (n = 17). In total, 20 gill samples originated from site A (10 with PGD1 and 10 with PGD 3, 10 samples from site B (7 with PGD1 and 3 with PGD 3) and 13 samples from site C (9 with PGD1 and 4 with PGD 3).
Project description:Amoebic Gill Disease (AGD), caused by the ectoparasite Paramoeba perurans (P. perurans) is characterised by hyperplasia of the gill epithelium and lamellar fusion and has become recognised as one of the most significant health threats in salmon farming . In this study, the gill and serum proteomes of Atlantic salmon inoculated with P. perurans, across multiple timepoints post-challenge, were analysed. The expression of proteins with established roles in innate immunity, across various timepoints, was compared with expression in naïve Atlantic salmon to elucidate the host response to gill colonisation.
Project description:The aim of this experiment was to explore transcriptomic changes in the gills of Altantic salmon (Salmo salar) following a model lab based parr smolt transformation from fresh water (FW) to salt water (SW). The process of smoltification (migration from FW to SW) is stimulated by long day photoperiod, which acts on the tissue-specific levels of active thyroid hormone (triiodothyronine, T3) through the expression of thyroid hormone type 2 deiodinase (dio2), responsible for conversion of inactive thyroxine (T4) to T3. To gain insight into the functional significance of dio2 induction, we performed a SW-challenge experiment in which we inhibited dio2 activity by addition of iopanoic acid (IOP) to the SW. We also assessed the ability of co-treatment with T3 to override IOP effects. Juvenile fish maintained in FW were subjected to a standard smolt photoperiod regime known to stimulate smoltification, after which they were randomly allocated to one of the four treatments: SW, SW+IOP, SW+IOP+T3 and FW as a control. Fish (n = 8-10 per treatment) were exposed to these conditions for 6 h and then sacrificed to obtain gill tissue for microarray analysis, carried out using a custom-designed Agilent oligonucleotide microarray platform Salar_2 (one glass slide with 4 x 44K arrays, Agilent design ID: 025520, array design A-MEXP-2065), developed and validated for Atlantic salmon (for details, see specific protocols) . In total, 36 hybridisations were performed on gills from individual fish, with 8-10 replicate fish per treatment. We identified 1939 genes whose expression was significantly increased or decreased by transfer from FW to SW. For a subset of 259 genes, this SW response was abolished if IOP was added to the SW, but maintained if T3 was also present during IOP treatment. This group of genes constitutes a candidate list, for which SW-inducibility appears to be dependent on locally mediated changes in gill T3 availability. The results of this experiment have been submitted for publication in Current Biology and are currently under review.
Project description:In aquaculture, recurrence rates of Amoebic Gill Disease (AGD), caused by the ectoparasite Paramoeba perurans (P. perurans) are high and no prophylactic strategies exist for disease prevention. In this study, Atlantic salmon (Salmo salar) were initially inoculated with P. perurans and following the development of amoebic gill disease were treated with fresh water immersion on day 21 and day 35 post inoculation. Fish were re-inoculated following a negative Q-PCR analysis for the presence of P. perurans. The gill host immune response was investigated at 7, 14 and 18 days post re-inoculation. Differential proteome expression of immune related proteins was assessed by comparison of each time point against naïve controls. In the gill, some proteins of the innate immune system were expressed in response to gill re-colonisation by P. perurans, while no features of adaptive immunity were found to be differentially expressed. Many of the proteins identified are novel in the context of AGD and their expression profiles suggest that their roles in the response to disease development and progression in single or multiple infections warrant further investigation.
Project description:Amoebic gill disease (AGD) is an ectoparasitic condition of some farm-reared marine fish and is caused by Neoparamoeba perurans. Tanks housing Atlantic salmon (Salmo salar) were inoculated with Neoparamoeba perurans and fish were sampled at 36 days postinoculation (pi.). AGD-affected gill tissue was dissected from N. perurans infected fish, and a DNA microarray was used to compare global gene expression against tissues from AGD-naive fish. To determine whether the changes in gene expression were restricted to AGD-lesions, lesion tissue from AGD-affected fish was also compared with non-lesion gill tissue dissected from the same gill arch. Samples were assessed using a DNA microarray. mRNA from lesion and non-lesion gill tissue was amplified and labeled. Six biological and two technical replicates were utilised to hybridise to 12 arrays using amplified RNA from AGD-affected lesion gill tissue with AGD-naive fish as a control. Four biological and two technical replicates were utilised to hybridise to 8 arrays using amplified RNA from AGD-affected lesion gill tissue with non-lesion tissue from the same gill arch as a control. The assignment of microarrays to treatment groups for hybridization was randomised by using a random number generator.
Project description:Klebsiella pneumoniae x546,ATCC15380, TMT,The samples were ground into a powder in liquid nitrogen. Then the powder was suspended in lysis buffer [1% sodium deoxycholate (SDS), 8M urea]. The mixture was allowed to settle at 4 鎺矯 for 30 min during which the sample were vortexed at every 5 min, and treated by ultrasound at 40 kHz and 40 W for 2 min. After centrifugation at 16000 rpm at 4鎺矯 for 30 min, the concentration of protein supernatant was determined by Bicinchoninic acid (BCA) method by BCA Protein Assay Kit (Pierce, Thermo, USA). Protein quantification was performed according to the kit protocol(Chen et al., 2021)
Project description:In this study we characterize the gill transcriptome changes in Gulf killifish (Fundulus grandis) that coincide with controlled laboratory-based exposure to various concentrations of experimentally-weathered south Louisiana crude oil. Gill transcription was contrasted between doses and across timepoints following dosing.
Project description:Prymnesium parvum is regarded as one of the most notorious harmful algal bloom (HAB) species worldwide. In recent years, it has frequently formed toxic blooms in coastal and brackish waters of America, Europe, Australia, Africa and Asia, causing large-scale mortalities of wild and cultured fish and other gill-breathing animals. In the last decade, blooms of P. parvum have expanded to inland fresh waters in the USA, presumably due to changes in environmental conditions. The aim of the experiment was to establish the gill transcriptomic responses to P. parvum in rainbow trout. We used 2 different concentrations of P. parvum and identified fish with low and moderate responses to the algae. Based on the dose of and the fish response, fish were classified into 4 groups with high exposure/moderate response (HM), high exposure/low response (HL), low exposure/low response (LL) and control group (C) with no exposure/no response. Gene expression profiling of the gill tissue was performed using a microarray platform developed and validated for rainbow trout.
Project description:Zinc deficiency is detrimental to organisms highlighting its role as an essential micronutrient contributing to numerous biological processes. To investigate the underlying molecular events invoked by zinc depletion we performed a temporal analysis of transcriptome changes observed within zebrafish gill. This tissue represents a model system for studying ion absorption across polarised cells as it provides a major pathway for fish to acquire zinc directly from water whilst sharing a conserved zinc transporting system with mammals. Zebrafish were treated with either zinc-depleted (water = 2.61 μg L-1; diet = 26 mg kg-1) or zinc-adequate (water = 16.3 μg L-1; diet = 233 mg kg-1) conditions for two weeks. Gill samples were collected at five time points and transcriptome changes analysed in quintuplicate using a 16K oligonucleotide array. Global transcript levels were measured in zebrafish gills using a oligonucleotide array either zinc-depleted or zinc-adequate diet. Gill samples were collected at five time points and transcriptome changes analysed in quintuplicate using a 16K oligonucleotide array
Project description:Long term chest blast exposure can lead to mental disorders, brain inflammation and oxidative stress in soldiers. However, the underlying mechanism of brain injury caused indirectly by chest blast remains unclear. It is urgent to find more potential biomarkers to reveal the deeper pathogenesis of this phenomenon. We used iTRAQ labeling combined with LC-MS/MS to screen potential differentially expressed proteins in rat brain after chest blast at different time points. Meanwhile, we also used Go, KEGG, David and Cytoscape to analyze the proteomic profile and explore its pathogenesis. Moreover, Western blotting was used to verify the target proteins. Our data showed that, a total of 6,931 proteins were identified. A total of 255 differentially expressed proteins were identified, of which 43, 84, 52, 97 and 49 proteins were identified from brain tissues at 12h, 24h, 48h, 72h, and 1w after chest blast exposure. Bioinformatics analysis, including GO, COG, KEGG and STRING, further proved that brain damage caused by chest blast exposure was involved in many important biological processes and signal pathways, such as inflammation, cell adhesion, phagocytosis, neuronal and synaptic damage, oxidative stress and apoptosis. Moreover, Western blotting further confirmed that these differentially expressed proteins and signaling pathways were associated with brain damage caused by chest blast exposure. For the first time, we screened and verified the potential protein biomarkers of brain damage caused indirectly by chest blast, and provided a new target for the treatment of this disease.