Gene expression profiling at the single nuclei level of channel catfish spleen
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ABSTRACT:
PROVIDER: PRJNA1114391 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR29111789.fastq.gz | Fastqsanger.gz | |||
| SRR29111790.fastq.gz | Fastqsanger.gz | |||
| SRR29111791.fastq.gz | Fastqsanger.gz | |||
| SRR29111792.fastq.gz | Fastqsanger.gz | |||
| SRR29111793.fastq.gz | Fastqsanger.gz |
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Project description:Channel catfish, Ictalurus punctatus, is an important model U.S. aquaculutre species. Given its role in food production, the catfish immune response to industry-relevant pathogens has been extensively studied and has provided crucial information on innate and adaptive immune function during disease progression. To further examine the channel catfish immune system, single-nuclei RNA sequencing on whole spleens from three adult individuals was performed. Spleen cell suspensions were prepared by passing tissues through cell sieves. Single-cell RNAseq libraries were then prepared using the 10X Genomics Chromium X with the Next GEM Single Cell 3’ Reagents and sequenced 2x150bp on an Illumina sequencer. Each demultiplexed library was aligned to the CoCo_2.0 channel catfish reference assembly, filtered, and counted to generate feature-barcode matrices. Cluster analysis allowed for the identification of multiple cell types including erythrocytes, hematopoietic stem cells, B cells, T/NK cells, myeloid lineage derivatives, plasma cells and endothelial cells. The majority of cells detected were erythrocytes, which are nucleated in teleost fish.
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Project description:Channel catfish, Ictalurus punctatus, are the primary cultured species of the catfish industry. Significant losses in production are attributed to bacterial diseases which are challenging to control and new interventions need to be developed. To gain insight into the in situ catfish immune system, we used single-nuclei RNAseq. We produced two single-nuclei atlases from three individual catfish using flow cytometry sorted IgM+ splenic cells. Libraries were prepared using the 10X Genomics system and sequenced on an Illumina sequencer. The reads were aligned to the reference genome (Ip Coco_2.0) using Cell Ranger (v8.0.0).
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Project description:Channel catfish and blue catfish represent two economically important freshwater aquaculture species in the United States. Our study aims to investigate the gene expression differences between these two catfish species by high-throughput RNA sequencing to understand their associated phenotypic differences in growth and disease resistant. Our transcriptomic analyses provide some insights into gene function differences between the two species and the molecular basis of channel catfish growth advantage in the tank culture environment.
2023-11-20 | GSE247918 | GEO
Project description:A high-density oligonucleotide microarray for channel catfish (Ictalurus punctatus) was designed and produced with Maskless Array Synthesizer technology (MAS). The microarray contained ~379,652 24-mer oligonucleotides covering ~18,999 catfish unique sequences. The global expression profiling of the catfish spleens stimulated by lipopolysaccharide (LPS) for 2h, 4h, 8h and 24h was investigated with the microarray. In the spleen samples, 409 genes were identified to be induced or repressed greater than 2-fold by LPS treatment. Self-organizing maps (SOM) clustering analysis was applied to interpret gene expression patterns, and 84 of these genes were clustered into six expression patterns. Real-time RT-PCR was used to verify the microarray results for 9 selected genes representing different expression levels. The results from real-time RT-PCR were positively correlated (R^2 = 0.87) with the results from the microarray. Channel catfish were injected with LPS or PBS carrier. Fish were killed by anesthesia overdose at 2, 4, 8, or 24h post-injection and total RNA was isolated from spleen. The experiment contained two replicate fish per time point. Relative signal intensities for each feature were generated using the Robust Multi-Array Average algorithm and log2 transformed. Transformed data were then processed using quantile normalization, and the average and std. error for each gene were calculated from the 10 perfectly matched oligos only. After detransformation, the ratio (fold change) was calculated using PBS treated samples (control) as the baseline against LPS-treated samples.
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Project description:The hybrid between female channel catfish (Ictalurus punctatus) and male blue catfish (Ictalurus furcatus) is superior in feed conversion, disease resistance, carcass yield, and harvestability compared to both parental species. However, heterosis and heterobeltiosis only occur in pond culture, and channel catfish grow much faster than the other genetic types in small culture units. This environment-dependent heterosis is intriguing, but the underlying genetic mechanisms are not well understood. In this study, phenotypic characterization and transcriptomic analyses were performed in the channel catfish, blue catfish, and their reciprocal F1s reared in tanks. The results showed that the channel catfish is superior in growth-related morphometrics, presumably due to significantly lower innate immune function, as investigated by reduced lysozyme activity and alternative complement activity. RNA-seq analysis revealed that genes involved in fatty acid metabolism/transport are significantly upregulated in channel catfish compared to blue catfish and hybrids, which also contributes to the growth phenotype. Interestingly, hybrids have a 40-80% elevation in blood glucose than the parental species, which can be explained by a phenomenon called transgressive expression (overexpression/underexpression in F1s than the parental species). A total of 1,140 transgressive genes were identified in F1 hybrids, indicating that 8.5% of the transcriptome displayed transgressive expression. Transgressive genes upregulated in F1s are enriched for glycan degradation function, directly related to the increase in blood glucose level. This study is the first to explore molecular mechanisms of environment-dependent hetero-sis/heterobeltiosis in a vertebrate species and sheds light on the regulation and evolution of heterosis vs. hybrid incompatibility.
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Project description:Seven early developmental stages in channel catfish, Ictalurus punctatus, were selected for transcriptome sequencing and analysis, Differential expression analysis and WGCNA approach was applied. The genes that play vital roles in embryogenesis and regulation of early development in channel catfish were detected. Our work reveals new insights for exploring the underlying mechanisms of channel catfish early development.
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Project description:A high-density oligonucleotide microarray for channel catfish (Ictalurus punctatus) was designed and produced with Maskless Array Synthesizer technology (MAS). The microarray contained ~379,652 24-mer oligonucleotides covering ~18,999 catfish unique sequences. The global expression profiling of the catfish spleens stimulated by lipopolysaccharide (LPS) for 2h, 4h, 8h and 24h was investigated with the microarray. In the spleen samples, 409 genes were identified to be induced or repressed greater than 2-fold by LPS treatment. Self-organizing maps (SOM) clustering analysis was applied to interpret gene expression patterns, and 84 of these genes were clustered into six expression patterns. Real-time RT-PCR was used to verify the microarray results for 9 selected genes representing different expression levels. The results from real-time RT-PCR were positively correlated (R^2 = 0.87) with the results from the microarray. Keywords: time course
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Project description:channel catfish (Ictalurus punctatus) Raw sequence reads
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Project description:Columnaris disease, caused by Flavobacterium covae, is one of three primary bacterial diseases that cause significant mortality and economic losses in farmed catfish (Ictalurus spp.). The quorum sensing gene luxR was reported to be upregulated in F. covae biofilm-grown cells stimulated with channel catfish (I. punctatus) mucus. To understand LuxR’s contributions to F. covae virulence, a F. covae C#2 luxR deletion mutant (ΔluxR) was created along with a complemented luxR (ΔluxR + pCE1). Channel catfish fingerlings challenged with the wild type or ΔluxR + pCE1 were observed to have 100% or 90% mortality, respectively; in contrast, only 2% mortality was observed for catfish fingerlings challenged with ΔluxR, indicating the important role for LuxR-regulated genes in F. covae virulence. A transcriptomic analysis indicated that LuxR-regulated genes in F. covae encode many functions including those associated with virulence, bacterial growth and metabolism, and survival during a fish infection. A decrease in biofilm formation was observed for ΔluxR relative to the wild-type and the complemented mutant, but only when mannose was included in the growth medium. The results of this study indicated that LuxR is a critical regulator of F. covae quorum sensing and virulence, warranting further research to identify and describe the relative contributions of LuxR-regulated virulence factors.
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