Golden cuttlefish Sepia esculenta Hoyle is an economically important cephalopod species. However, artificial hatching is currently challenged by low survival rate of larvae due to abnormal embryonic development. Dissecting the genetic foundation and regulatory mechanisms in embryonic development requires genomic background knowledge. Therefore, we carried out a transcriptome sequencing on Sepia embryos and larvae via mRNA-Seq. 32,597,241 raw reads were filtered and assembled into 98,615 unigenes (N50 length at 911 bp) which were annotated in NR database, GO and KEGG databases respectively. Digital gene expression analysis was carried out on cleavage stage embryos, healthy larvae and malformed larvae. Unigenes functioning in cell proliferation exhibited higher transcriptional levels at cleavage stage while those related to animal disease and organ development showed increased transcription in malformed larvae. Homologs of key genes in regulatory pathways related to early development of animals were identified in Sepia. Most of them exhibit higher transcriptional levels in cleavage stage than larvae, suggesting their potential roles in embryonic development of Sepia. The de novo assembly of Sepia transcriptome is fundamental genetic background for further exploration in Sepia research. Our demonstration on the transcriptional variations of genes in three developmental stages will provide new perspectives in understanding the molecular mechanisms in early embryonic development of cuttlefish.

The East Asian common octopus (Octopus sinensis) is an economically important species among cephalopods. This species exhibits a strict dioecious and allogamous reproductive strategy, along with a phenotypic sexual dimorphism, where the third right arm differentiates into hectocotylus in males. However, our understanding of the molecular mechanisms that underlie sex determination and differentiation in this species remains limited. In the present study, we surveyed gene-expression profiles in the immature male and female gonads of O. sinensis based on the RNA-seq, and a total of 47.83 Gb of high-quality data were generated. Compared with the testis, we identified 8302 differentially expressed genes (DEGs) in the ovary, of which 4459 genes were up-regulated and 3843 genes were down-regulated. Based on the GO enrichment, many GO terms related to sex differentiation were identified, such as sex differentiation (GO: 0007548), sexual reproduction (GO: 0019953) and male sex differentiation (GO: 0046661). A KEGG classification analysis identified three conserved signaling pathways that related to sex differentiation, including the Wnt signaling pathway, TGF-β signaling pathway and Notch signaling pathway. Additionally, 21 sex-related DEGs were selected, of which 13 DEGs were male-biased, including Dmrt1, Foxn5, Foxj1, Sox30, etc., and 8 DEGs were female-biased, including Sox14, Nanos3, β-tubulin, Suh, etc. Ten DEGs were used to verify the expression patterns in the testis and ovary using the RT-qPCR method, and the results showed that the expression level shown by RT-qPCR was consistent with that from the RNA-seq, which confirmed the reliability of the transcriptome data. The results presented in this study will not only contribute to our understanding of sex-formation mechanisms in O. sinensis but also provide the foundational information for further investigating the molecular mechanisms that underline its gonadal development and facilitate the sustainable development of octopus artificial breeding.

The study aimed to uncover the molecular response of juvenile Sebastes schlegelii to air exposure stress by identifying differentially expressed genes (DEGs) that may underlie their anti-stress mechanisms. Juvenile Sebastes schlegelii were subjected to varying durations of air exposure stress. The total RNA was extracted from whole tissues and sequenced using the Illumina NovaSeq 6000 platform. The transcriptome data were analyzed to identify DEGs through pairwise comparisons across a control group and two experimental groups exposed to air for 40 s and 2 min 30 s, respectively. The comparative DEG analysis revealed a significant number of transcripts responding to air exposure stress. Specifically, 5173 DEGs were identified in the 40 s exposure group (BS) compared to the control (BC), 6742 DEGs in the 2 min 30 s exposure group (BD) compared to the control (BC), and 2653 DEGs when comparing the BD to the BS group. Notably, Gene Ontology (GO) analysis showed an enrichment of DEGs associated with peptidase activity and extracellular regions, suggesting a role in the organism’s stress response. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis pointed to the involvement of metabolic pathways, which are crucial for energy management under stress. The upregulation of protein digestion and absorption pathways may indicate a physiological adaptation to nutrient scarcity during stress. Additionally, the identification of antibiotic biosynthesis pathways implies a potential role in combating stress-induced infections or damage. The identified DEGs and enriched pathways provide insights into the complex anti-stress response mechanisms in juvenile Sebastes schlegelii. The enrichment of peptidase activity and extracellular region-related genes may reflect the initiation of tissue repair and immune response following air exposure. The connection between protein digestion and absorption pathways and anti-stress capabilities could be interpreted as a metabolic readjustment to prioritize energy-efficient processes and nutrient assimilation during stress. The role of antibiotic biosynthesis pathways suggests a defensive mechanism against oxidative stress or microbial invasion that might occur with air exposure.

Objectives: Acute temperature stress was explored in Hippocampus abdominalis through a comprehensive RNA-seq analysis. Methods: RNA-seq was conducted on 20-day-old H. abdominalis after 24 h of temperature stress. Four experimental conditions were established: a control group (18 °C) and three temperature treatment groups (21, 24, and 27 °C). Results: Seahorse larvae were found to be unaffected by 21 °C and 24 °C and were able to survive for short periods of time during 24 h of incubation, whereas mortality approached 50% at 27 °C. The sequencing process produced 75.63 Gb of high-quality clean data, with Q20 and Q30 base percentages surpassing 98% and 96%, respectively. A total of 141, 333, and 1598 differentially expressed genes were identified in the 21, 24, and 27 °C groups vs. a control comparison group, respectively. Notably, the number of up-regulated genes was consistently higher than that of down-regulated genes across all comparisons. Gene Ontology functional annotation revealed that differentially expressed genes were predominantly associated with metabolic processes, redox reactions, and biosynthetic functions. In-depth KEGG pathway enrichment analysis demonstrated that down-regulated genes were significantly enriched in pathways related to steroid biosynthesis, terpenoid backbone biosynthesis, spliceosome function, and DNA replication. Up-regulated genes were enriched in pathways associated with the FoxO signaling pathway and mitophagy (animal). The results indicated that temperature stress induced extensive changes in gene expression in H. abdominalis, involving crucial biological processes such as growth, biosynthesis, and energy metabolism. Conclusions: This study provided key molecular mechanisms in the response of H. abdominalis to temperature stress, offering a strong basis for future research aimed at understanding and mitigating the effects of environmental stressors on marine species.

Flesh texture is one of the most important quality traits determining consumer satisfaction and perception of fish. Traditional methods for measuring flesh texture characteristics present several limitations. This study developed a rapid, efficient and non-invasive approach based on hyperspectral imaging (HSI) technology for evaluating texture profile analysis (TPA) parameters in live spotted seabass. Three types of surface hyperspectral data (skin with scales, SWS; skin without scales, SOS; and reversed skin without scales, RSOS) were collected from the dorsal region of 300 live fish across 400–1000 nm wavelength range. After applying five spectral pre-processing methods, five machine learning algorithms, including partial least squares regression (PLSR), least square support vector machine regression (LS-SVR), random forest (RF), convolutional neural network (CNN) and back propagation artificial neural network (BP-ANN), were performed to construct optimal prediction models for seven TPA parameters. Gumminess was predicted most effectively, with the RF model achieving a prediction set coefficient of determination (R²_P) of 0.923, a ratio of performance deviation (RPD) of a mean absolute percentage error (MAPE) of 6.378%. The optimal models enabled quantitative prediction of gumminess, chewiness, and hardness (RPD > 3.0 and MAPE<10%). For cohesiveness and springiness, the models performed satisfactory with R²_P values of 0.859 and 0.854, RPD values of 2.890 and 2.613, and MAPE were 6.858% and 3.002%, respectively. However, prediction accuracy for adhesiveness and resilience was lower, with R²_P values of 0.552 and 0.341, and RPD values of 1.494 and 1.134, respectively. Visualization maps generated from the optimal models facilitated the direct assessment of TPA parameters distribution in the muscle. This study demonstrated that HSI combined with machine learning offers an effective and non-invasive method for assessing flesh quality in live spotted seabass, thereby accelerating the selection of germplasm with superior textural attributes.