Feamp_MiSS
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB80705 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR13732100.fastq.gz | Fastqsanger.gz | |||
| ERR13732101.fastq.gz | Fastqsanger.gz | |||
| ERR13732102.fastq.gz | Fastqsanger.gz | |||
| ERR13732103.fastq.gz | Fastqsanger.gz | |||
| ERR13732104.fastq.gz | Fastqsanger.gz |
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Project description:Circulating plasma microRNAs (miRNAs) are well established as biomarkers of several diseases in humans and have recently been used as indicators of environmental exposures in fish. However, the role of plasma miRNAs in regulating acute stress responses in fish is largely unknown. Tissue and plasma miRNAs have recently been associated with excreted miRNAs in humans however external miRNAs have never been measured in fish. The objective of this study was to characterize the plasma miRNA profile in response to acute stress in rainbow trout (Oncorhynchus mykiss), as well as miRNA profiles in novel external samples, (fish epidermal mucus and the surrounding water). RNA was extracted and sequenced from plasma, mucus, and water collected from rainbow trout and their surrounding environment prior to and one-hour following a three-minute air exposure, a known inducer of an acute stress response in fish. Following small RNA-Seq and pathway analysis, we identified differentially expressed plasma miRNAs that targeted biosynthetic, degradation, and metabolic pathways. We successfully isolated miRNA from trout mucus and the surrounding water and detected differences in miRNA expression one-hour post air stress. The altered miRNA profiles in mucus and water were unique to the altered plasma miRNA profile, indicating that the plasma miRNA response was not associated with or immediately reflected in external samples. This research expands our understanding of the role of plasma miRNA in the acute stress response of fish and is the first study to report on the successful isolation and profiling of miRNA from fish mucus and water samples. Measurements of miRNA from plasma, mucus, and water can be further studied and have the potential to be applied in environmental monitoring as non-lethal indicators of acute stress in fish.
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Project description:The kiss system is known to regulate puberty and to be involved in the onset of reproduction, acting upstream of the Brain-Pituitary-Gonad axis in gonadotrophin releasing hormone neurons and stimulating secretion of GnRH. Kisspeptin has been proven to be a novel therapeutic for fertility disorders in humans. In fish, positive results have also observed the prospective of this hormone to induce gamete development. During the last two decades, support has been obtained describing the key role of microRNAs (miRNA) in regulating gene expression at the post-transcriptional level and they have been recommended as relevant biomarkers in reproductive studies. Senegalese sole is a marine flatfish species with high importance in the European aquaculture industry, due to its good growth rate and high market value. However, this species presents a bottleneck in terms of reproduction, which does not allow to close the cycle in captive conditions. Hence, this study aimed to observe the effect of a novel hormonal treatment using kisspeptin in miRNA expression related to reproductive performance in cultured Senegalese sole breeders. For this purpose, Senegalese sole F1 broodstock were held at CCMAR facilities in 4 tanks of 3m3 (n = 12/per tank, sex ratio 1:1) under natural photoperiod and temperature conditions. During the reproductive season, the animals from each 2 of the tanks were treated either with SSkiss2_v2 decapeptide in a single intramuscular injection, or with a PBS injection for control groups. After the treatment (2 and 4 days), all fish were anesthetized and sampled for blood collection. Afterwards, plasma was obtained to determine several parameters related to reproductive physiology (Fsh, Lh and sexual esteroids (T, 11-KT and E2)) and miRNAs regulation. After observing the high implication of this treatment in the gonadotrofins, above all in Fsh and sexual esteroids, above all in T, the RNA was extracted from blood plasma using miRNeasy Serum/Plasma Kit (Qiagen, Germany) and libraries were prepared using NEXTflex Small RNA-Seq kit v3 (Bioo Scientific, USA). Small RNA sequencing (sRNA-seq) (NextSeq 500) was performed on sole treated with kisspeptin (n = 16) and non-treated control fish (n = 14). A total of 1,467 unique miRNAs were expressed in Senegalese sole plasma. A multidimensional scaling (MDS) plot showed no clear clustering of the samples with the kisspeptin treatment. However, the differential expression analysis revealed one miRNA, miR-1-3p, to be highly up-regulated in both males and females treated with kisspeptin compared to control sole. This miRNA is linked with myogenesis and angiogenesis, and skeletal muscle development in fish. Three isomirs (let 7e, miR-199a-3p, miR-100-5p) were differentially expressed between females treated and control fish. MicroRNA let-7e is an important regulator of endothelial function and inflammation, related to testosterone pathway release; miR-199a-3p plays an important role in inflammation and women fertility and miR-100-5p might be implicated in the regulation of cell growth and oocytes maturation. In conclusion, kisspeptin treatment may affect gonad maturation in females and muscle growth in males. Our data support the possibility of using circulating miRNAs as less invasive physiological biomarkers in fish reproduction.
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Project description:Pistacia chinensis Bunge is known as dioecious, but we have found wild monoecious individuals. In order to screen the candidate genes which may influence the sex expression or floral phenotypic differences of P. chinensis, the inflorescence buds for different sex types associated with the sex differentiation were selected and tested for small RNA sequencing. Sex-specific differentially expressed small RNA were discovered, combined with real-time PCR data, the regulation patterns of various sex types were first revealed. Our study represents the first detailed analysis of small RNA sequencing, providing more clues for understanding the mechanism of sex determination on P. chinensis.
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