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Leptin is a peptide hormone primarily produced by adipose tissue, which plays a crucial role in regulating energy balance by controlling appetite and energy expenditure. To better understand the intricate physiological functions of leptin in hepatocytes in tongue sole (Cynoglossus semilaevis), this study presents an integrated transcriptomic and metabolomic analysis of tongue sole hepatocytes following stimulation with lepA and lepB. Comparative analysis identified 2971 and 2900 upregulated differentially expressed genes (DEGs) and 1895 and 1821 downregulated DEGs in response to lepA and lepB stimulation, respectively. Notably, 5706 genes were commonly regulated by both stimulations, suggesting overlapping functional roles of these two leptin proteins. GO and KEGG enrichment analysis indicated significant enrichment in immune response (JAK-STAT signaling pathway, NF-κB signaling pathway, etc.) and lipid metabolism pathways, such as fatty acid synthesis, with similar findings for lepB. Metabolomic analysis demonstrated clear separation between treatment and control groups, with 34 medium- to long-chain fatty acids and numerous differentially expressed metabolites (DEMs) identified. KEGG analysis of the metabolome paralleled the transcriptomic findings, with shared pathways in lipid metabolism and immune function. Finally, joint analysis highlighted co-enrichment in linoleic acid metabolism and leishmaniasis pathways. Detailed mechanistic insights revealed the activation of JAK-STAT and NF-κB signaling pathways, modulation of arachidonic acid metabolism, and the influence on the MAPK signaling pathway. Additionally, lepA uniquely co-enriched in the fatty acid synthesis pathway, with specific gene regulation affecting the synthesis of various fatty acids. The study concluded that lepA and lepB exerted significant regulatory effects on lipid metabolism and immune responses in tongue sole hepatocytes through complex molecular networks. To our current knowledge, this represents the first direct evidence of leptin's role in immune function within teleost fish and offers valuable insights into the molecular mechanisms of lipid metabolism underlying the actions of lepA and lepB.

Sea-land relay transportation (SRT) is a key part of the “land-based greenhouse + offshore net cage” mariculture system for yellowtail kingfish. In this study, the physiological status of yellowtail kingfish during SRT was assessed by determining serum biochemical indexes (levels of cortisol, glucose, and malondialdehyde (MDA), and activities of superoxide dismutase (SOD) and catalase (CAT)), the composition of intestinal microflora, and the liver transcriptome before (control), during, and after transportation (T1 and T2) and after recovery for 6 hours (6 h), 12 hours (12 h), 1 day (D1), 3 days (D3), and 7 days (D7). The results show that the serum cortisol and glucose levels, MDA level, and activities of SOD and CAT significantly increased during transportation, and decreased to similar levels to those in the control after 1 day of recovery. Bacteria including Acinetobacter, Enterobacter, and Comamonas accounted for more than 91 % of the microbial community in the gut of yellowtail kingfish, and these genera were associated with changes in serum glucose levels, indicating that they may help to alleviate the effects of transportation stress. Analyses of liver transcriptome data showed that significantly differentially expressed genes (DEGs) in the T2, D1, D3 and D7 groups were significantly enriched in the “Protein processing in the endoplasmic reticulum (ER)” pathway. At the end of transportation (T2 group), the ER-associated degradation (ERAD) pathway was activated, with up-regulation of hsp40, hsp70, and hsp90α. The transcript levels of these genes gradually returned to levels similar to those in the control during the recovery period. At the first day of recovery, the unfolded protein response imposed stress upon the ER, accompanied by significant down-regulation of PEPK and Calmin and significant up-regulation of CRT. The DEGs in the D3 group were enriched in the “ER”, “Steroid biosynthesis”, “Fatty acid degradation”, “PPAR signaling”, and “Fat digestion and absorption” pathways, whereas the number of DEGs related to lipid metabolism and stress pathways was significantly decreased in the D7 group. Our results indicate that hsp70 and hsp90α can be used as stress markers, and hsp40 can be used as a physio-metabolic recovery marker for yellowtail kingfish. These results show that, although serum indices recovered 24 hours after transportation, up to 3 days after transportation is an important period for recovery from transportation stress at the transcriptome level. “Protein processing in the ER pathway” is an important pathway regulating stress and recovery in yellowtail kingfish.

The neurokinin B (NKB) system acts as a crucial modulator in the regulation of reproduction and feeding in mammals, however limited information is available in teleosts regarding its potential role in reproductive functions. Herein, we have identified its encoding gene, namely tachykinin 3 (tac3), in the yellowtail kingfish. The full-length tac3 cDNA sequence was 372 bp in size and encoded a 123-aa preprohormone, including two mature peptides, NKB and NKF. Tissue distribution analysis showed that tac3 was primarily expressed in the intestine of yellowtail kingfish. During embryonic development, the highest mRNA levels of tac3 were observed at stage 4 (16-cell) and stage 14 (newly-hatched larva). The peak of tac3 transcript levels was also noticed at 30 days post-hatching during early ontogeny. The physiological effects of NKB and NKF on expression of brain and pituitary genes along with plasma hormones levels were further analyzed by intraperitoneal injection. NKB exerted a stimulatory effect on gnrh1, gnrh2, gnih, and kiss2r mRNA levels, but suppressed kiss1r, gnihr, gal, gh, lhβ, and fshβ mRNA levels. An evident increase was observed on transcript levels of gnrh1, gnrh2, kiss1, kiss2, gnih, and gnihr after NKF treatment, whereas kiss1r, kiss2r, gh, lhβ, and fshβ mRNA levels were down-regulated. Moreover, NKF, but not NKB, reduced serum estradiol levels, and both of these two peptides suppressed serum 11-keto testosterone levels. Taken together, our results provide preliminary evidence that the NKB system participates in the control of reproduction in yellowtail kingfish via acting on brain, pituitary and gonadal hormones.

This study investigated the physiological responses to copper toxicity in liver and gill of juvenile yellowtail kingfish by exposing to 0 (control), 0.0672, 0.12, 0.214, 0.384, 0.672, 1.20, or 2.14 mg/L copper sulfate for 7 days and then recovered for 14 days. The histopathology, enzyme activities and differential gene expression were detected. The results indicate the safe concentration value of copper for juvenile yellowtail kingfish is 0.083 mg/L. Copper exposure caused structural damage to gill and liver, and could be disappeared or restored after the recovery period. Copper exposure enhanced the Cu,Zn superoxide dismutase (Cu/Zn-SOD) and glutathione peroxidase (GSH-Px) activities and the malondialdehyde (MDA) level in liver. However, with increasing copper concentrations, liver aspartate transaminase (AST) and gill Na+/K+-ATPase activities were at first enhanced and then inhibited. Serum lysozyme (LZM) and Cu/Zn-SOD activities were enhanced at low copper concentrations but inhibited at high concentrations, and the activities of alanine transaminase (ALT) and AST significantly increased at greater copper concentrations. After the recovery period, only liver Cu/Zn-SOD activity returned to control level. In contrast, liver GSH-Px activity and MDA levels and gill Na+/K+-ATPase activity, serum LZM and AST activities remained significantly higher than control, but liver AST activity, serum ALT, and Cu/Zn-SOD activities were significantly lower than control. We constructed the gene regulatory networks of molecular pathways in liver and gill in response to copper exposure. The DEGs were enriched mainly in the pathway of lipid metabolism in liver and pathway of oxidative phosphorylation in gill, indicating these respective pathways are involved in the response to copper stress in this species. The present results elucidate some histological, physiological, and molecular mechanisms underlying copper-exposure stresses in yellowtail kingfish, which could help guide the safe utilization of copper sulfate in disease control of farmed individuals.

Ice, Cloud, and Land Elevation Satellite-2 (ICESat-2) has shown great potential for near-shore bathymetry. Unavoidably, the photon data acquired has a lot of noise. In order to remove noise more accurately, a photon denoising algorithm based on adaptive elevation difference thresholding (AEDTA) is proposed under the assumption that the terrain is continuously changing. The algorithm uses a Gaussian function obtained by fitting an elevation histogram to ICESat-2 data for the extraction of underwater photons. Repeatedly count the elevation difference histogram of underwater photons and adaptively determine the threshold. And the final denoised underwater signal photons are obtained. Compared with the conventional clustering method of ordering points to identify the clustering structure (OPTICS), our algorithm achieves better photon detection results for underwater topography for different densities of ICESat-2 data and has the advantage of parameter adaption without human intervention. ICESat-2 data from Ganquan Island, Dongdao Island and Oahu Island were selected for testing, and the ICESat-2 bathymetry results calculated by the algorithm were compared with in situ bathymetry data to assess the reliability and accuracy of the proposed denoising algorithm. The results show that the MAE of the bathymetry results calculated using the ICESat-2 signal photon data extracted by the method ranged from 0.54 to 1.57 m and the RMSE ranged from 0.64 to 1.74 m in different seas. The algorithm can provide high quality denoised data for bathymetric mapping around coastal areas and islands. Meanwhile, the AEDTA algorithm also has a good denoising effect on terrestrial photons, and the effect of cloud shading on the Gaussian fit analysis of ICESat-2 data cannot be ignored.

Although recent studies have revealed its involvement in fish reproduction, the physiological role of neurokinin B (NKB), which is encoded by the tachykinin 3 (tac3) gene, is still largely unclear in teleosts, with most researches on the orders Perciformes and Cypriniformes. In the current study we identified the tac3 gene in a representative species of Pleuronectiformes, the half-smooth tongue sole. The full-length cDNA sequence of tongue sole tac3 was 742 base pairs (bp) in length with an open reading frame of 372 bp that encoded a 123-amino acids preprohormone. The TAC3 precursor contained NKB and an additional peptide NKB-related peptide (NKBRP), both of which had an –FVGLM motif at the C-termini. Tissue distribution analysis indicated that tac3 transcripts were highly expressed in the brain, intestine and gonads of both sexes, and to a lesser extent in other tissues. Among various brain regions, tac3 transcripts were most abundant in the hypothalamus. In addition, the expression profiles of tac3 mRNA were also evaluated along the brain-pituitary-gonadal axis during tongue sole ovarian maturation. The brain tac3 mRNA levels remained unchanged, however, in both pituitary and ovary, the mRNA expression of tac3 increased significantly to the maximal levels at stage III and then declined back to basal levels. Taken together, our results encompass the first study reporting the existence of tac3 gene in a flatfish species and suggest its possible involvement in the control of reproduction in the half-smooth tongue sole.

Spexin (SPX) is a newly identified peptide hormone of 14 amino acids (aa) with pleiotropic functions in vertebrates, and recently a second paralog of SPX (named SPX2) has been only found in some non-mammalian species (the previous form of SPX is now designated as SPX1). Results of previous studies have revealed the existence of a functional SPX1 in the half-smooth tongue sole. To clarify the divergence of the spexin family, we further isolated the spx2 gene from the same flatfish species in the current study. The open reading frame of tongue sole spx2 was 378 nucleotides in size that encoded a 125-aa preprohormone, and the tongue sole SPX2 precursor included a 17-aa mature peptide that may be amidated at the C-terminus, rather than a tetradecapeptide reported in most other vertebrates. Tissue distribution analysis showed that spx2 transcripts were found in a wide array of tissues of both sexes with the highest expression in the ovary. In addition, the spx2 mRNA abundance was also ubiquitously detected in different brain regions. Food deprivation for 4 weeks stimulated spx2 mRNA levels at the brain-pituitary-ovary axis, and refeeding of starved fish restored the spx2 mRNA levels back to normal. Taken together, this study provides initial evidence for the existence of a novel spexin gene (spx2) in Pleuronectiformes and suggest its possible involvement in the regulation of energy balance in the female tongue sole.