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Insights Into Sexual Maturation and Reproduction in the Norway Lobster (Nephrops norvegicus) via in silico Prediction and Characterization of Neuropeptides and G Protein-coupled Receptors.

ABSTRACT: Multiple biological processes across development and reproduction are modulated by neuropeptides that are predominantly produced and secreted from an animal's central nervous system. In the past few years, advancement of next-generation sequencing technologies has enabled large-scale prediction of putative neuropeptide genes in multiple non-model species, including commercially important decapod crustaceans. In contrast, knowledge of the G protein-coupled receptors (GPCRs), through which neuropeptides act on target cells, is still very limited. In the current study, we have used in silico transcriptome analysis to elucidate genes encoding neuropeptides and GPCRs in the Norway lobster (Nephrops norvegicus), which is one of the most valuable crustaceans in Europe. Fifty-seven neuropeptide precursor-encoding transcripts were detected, including phoenixin, a vertebrate neurohormone that has not been detected in any invertebrate species prior to this study. Neuropeptide gene expression analysis of immature and mature female N. norvegicus, revealed that some reproduction-related neuropeptides are almost exclusively expressed in immature females. In addition, a total of 223 GPCR-encoding transcripts were identified, of which 116 encode GPCR-A (Rhodopsin), 44 encode GPCR-B (Secretin) and 63 encode other GPCRs. Our findings increase the molecular toolbox of neural signaling components in N. norvegicus, allowing for further advances in the fisheries/larvae culture of this species.

SUBMITTER: Nguyen TV

PROVIDER: S-EPMC6073857 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundThe Norway lobster, Nephrops norvegicus, is economically important in European fisheries and is a key organism in local marine ecosystems. Despite multi-faceted scientific interest in this species, our current knowledge of genetic resources in this species remains very limited. Here, we generated a reference de novo transcriptome for N. norvegicus from multiple tissues in both sexes. Bioinformatic analyses were conducted to detect transcripts that were expressed exclusively in either males or females. Patterns were validated via RT-PCR.ResultsSixteen N. norvegicus libraries were sequenced from immature and mature ovary, testis and vas deferens (including the masculinizing androgenic gland). In addition, eyestalk, brain, thoracic ganglia and hepatopancreas tissues were screened in males and both immature and mature females. RNA-Sequencing resulted in >600 million reads. De novo assembly that combined the current dataset with two previously published libraries from eyestalk tissue, yielded a reference transcriptome of 333,225 transcripts with an average size of 708 base pairs (bp), with an N50 of 1272 bp. Sex-specific transcripts were detected primarily in gonads followed by hepatopancreas, brain, thoracic ganglia, and eyestalk, respectively. Candidate transcripts that were expressed exclusively either in males or females were highlighted and the 10 most abundant ones were validated via RT-PCR. Among the most highly expressed genes were Serine threonine protein kinase in testis and Vitellogenin in female hepatopancreas. These results align closely with gene annotation results. Moreover, a differential expression heatmap showed that the majority of differentially expressed transcripts were identified in gonad and eyestalk tissues. Results indicate that sex-specific gene expression patterns in Norway lobster are controlled by differences in gene regulation pattern between males and females in somatic tissues.ConclusionsThe current study presents the first multi-tissue reference transcriptome for the Norway lobster that can be applied to future biological, wild restocking and fisheries studies. Sex-specific markers were mainly expressed in males implying that males may experience stronger selection than females. It is apparent that differential expression is due to sex-specific gene regulatory pathways that are present in somatic tissues and not from effects of genes located on heterogametic sex chromosomes. The N. norvegicus data provide a foundation for future gene-based reproductive studies.

Project description:Underwater video monitoring systems are being widely used in fisheries to investigate fish behavior in relation to fishing gear and fishing gear performance during fishing. Such systems can be useful to evaluate the catch composition as well. In demersal trawl fisheries, however, their applicability can be challenged by low light conditions, mobilized sediment and scattering in murky waters. In this study, we introduce a novel observation system (called NepCon) which aims at reducing current limitations by combining an optimized image acquisition setup and tailored image analyses software. The NepCon system includes a high-contrast background to enhance the visibility of the target objects, a compact camera and an artificial light source. The image analysis software includes a machine learning algorithm which is evaluated here to test automatic detection and count of Norway lobster (Nephrops norvegicus). NepCon is specifically designed for applications in demersal trawls and this first phase aims at increasing the accuracy of N. norvegicus detection at the data acquisition level. To find the best contrasting background for the purpose we compared the output of four image segmentation methods applied to static images of N. norvegicus fixed in front of four test background colors. The background color with the best performance was then used to evaluate computer vision and deep learning approaches for automatic detection, tracking and counting of N. norvegicus in the videos. In this initial phase we tested the system in an experimental setting to understand the feasibility of the system for future implementation in real demersal fishing conditions. The N. norvegicus directed trawl fishery typically has no assistance from underwater observation technology and therefore are largely conducted blindly. The demonstrated perception system achieves 76% accuracy (F-score) in automatic detection and count of N. norvegicus, which provides a significant elevation of the current benchmark.

Dataset Information

Insights Into Sexual Maturation and Reproduction in the Norway Lobster (Nephrops norvegicus) via in silico Prediction and Characterization of Neuropeptides and G Protein-coupled Receptors.

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