Pub Date : 2025-08-18DOI: 10.1016/j.cbpb.2025.111146
Xishuang Shan , Xiya Zhang , Binbin Tao , Yanlong Song , Zuoyan Zhu , Wei Hu , Ji Chen
Gonadal development and gamete maturation are essential for fish reproduction. The protein tyrosine phosphatase receptor type Fb (Ptprfb) is a member of the tyrosine phosphatase family. In the present study, we used CRISPR/Cas9 to mutate ptprfb in zebrafish. A significantly reduced natural fertilization rate of sperm from mutant fish was observed. The mutant fish produced fewer sperm with shorter flagella, and a smaller proportion of sperm could be activated. RNA-seq analysis revealed abnormal expression of some genes in testicular cells, such as hemoglobin subunit βA1 (hbba1), myosin heavy chain 11b (myh11b), and transgelin (tagln), as well as some genes involved in focal adhesion formation. These findings demonstrate that ptprfb contributes to spermatogenesis in zebrafish, and its dysfunction can adversely affect both the quantity and quality of sperm.
{"title":"CRISPR/Cas9-mediated editing of ptprfb (protein tyrosine phosphatase receptor type fb) reveals its regulatory role in zebrafish spermatogenesis","authors":"Xishuang Shan , Xiya Zhang , Binbin Tao , Yanlong Song , Zuoyan Zhu , Wei Hu , Ji Chen","doi":"10.1016/j.cbpb.2025.111146","DOIUrl":"10.1016/j.cbpb.2025.111146","url":null,"abstract":"<div><div>Gonadal development and gamete maturation are essential for fish reproduction. The protein tyrosine phosphatase receptor type Fb (Ptprfb) is a member of the tyrosine phosphatase family. In the present study, we used CRISPR/Cas9 to mutate <em>ptprfb</em> in zebrafish. A significantly reduced natural fertilization rate of sperm from mutant fish was observed. The mutant fish produced fewer sperm with shorter flagella, and a smaller proportion of sperm could be activated. RNA-seq analysis revealed abnormal expression of some genes in testicular cells, such as hemoglobin subunit βA1 (<em>hbba1</em>), myosin heavy chain 11b (<em>myh11b</em>), and transgelin (<em>tagln</em>), as well as some genes involved in focal adhesion formation. These findings demonstrate that <em>ptprfb</em> contributes to spermatogenesis in zebrafish, and its dysfunction can adversely affect both the quantity and quality of sperm.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111146"},"PeriodicalIF":1.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-18DOI: 10.1016/j.cbpb.2025.111147
Won Chae Jeong , Kun Woo Kim , Jin A Kim , Jun-Hwan Kim , Cheol Young Choi
Microplastics and trace metals such as cadmium (Cd) are environmental contaminants commonly co-occurring in marine ecosystems. We aimed to evaluate the impact of combined exposure of Pacific oyster (Crassostrea gigas) to microbeads (MBs) and Cd, focusing on the effects of the depuration process on contaminant removal and stress-related biomarkers. Pacific oysters were exposed to MBs, Cd, and their combination for 48 h, followed by a 72 h depuration process using uncontaminated seawater. We measured the levels of accumulated MBs and Cd in the whole soft tissue of the Pacific oysters to evaluate the degree of contaminant removal. Additionally, the concentrations of hydrogen peroxide were measured and the mRNA expression levels of antioxidant enzymes, metallothionein, and the apoptosis-related gene caspase-3 were analyzed in the Pacific oyster hepatopancreas tissue to evaluate oxidative stress and apoptosis. Our results indicated that Cd was eliminated more slowly than MBs, and the Pacific oysters exposed to combined MB and Cd contaminants maintained higher levels of oxidative stress-related gene expression than those exposed to individual contaminants. These findings suggest that Cd may persist longer in oyster tissues than MBs, potentially leading to prolonged toxicity in the Pacific oyster. Furthermore, in environments where both MBs and Cd are present, MBs can enhance the toxic effects of Cd through a synergistic interaction. Overall, we provide a reference for understanding the depuration and physiological responses of marine bivalves exposed to MBs and Cd.
{"title":"Depuration and post-exposure recovery of oxidative stress responses to microplastics and cadmium in Pacific oyster (Crassostrea gigas)","authors":"Won Chae Jeong , Kun Woo Kim , Jin A Kim , Jun-Hwan Kim , Cheol Young Choi","doi":"10.1016/j.cbpb.2025.111147","DOIUrl":"10.1016/j.cbpb.2025.111147","url":null,"abstract":"<div><div>Microplastics and trace metals such as cadmium (Cd) are environmental contaminants commonly co-occurring in marine ecosystems. We aimed to evaluate the impact of combined exposure of Pacific oyster (<em>Crassostrea gigas</em>) to microbeads (MBs) and Cd, focusing on the effects of the depuration process on contaminant removal and stress-related biomarkers. Pacific oysters were exposed to MBs, Cd, and their combination for 48 h, followed by a 72 h depuration process using uncontaminated seawater. We measured the levels of accumulated MBs and Cd in the whole soft tissue of the Pacific oysters to evaluate the degree of contaminant removal. Additionally, the concentrations of hydrogen peroxide were measured and the mRNA expression levels of antioxidant enzymes, metallothionein, and the apoptosis-related gene caspase-3 were analyzed in the Pacific oyster hepatopancreas tissue to evaluate oxidative stress and apoptosis. Our results indicated that Cd was eliminated more slowly than MBs, and the Pacific oysters exposed to combined MB and Cd contaminants maintained higher levels of oxidative stress-related gene expression than those exposed to individual contaminants. These findings suggest that Cd may persist longer in oyster tissues than MBs, potentially leading to prolonged toxicity in the Pacific oyster. Furthermore, in environments where both MBs and Cd are present, MBs can enhance the toxic effects of Cd through a synergistic interaction. Overall, we provide a reference for understanding the depuration and physiological responses of marine bivalves exposed to MBs and Cd.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111147"},"PeriodicalIF":1.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kuruma shrimp (Penaeus japonicus) exhibits sand-diving behaviour. The genetic control mechanism of this behaviour is still unclear, although previous single-cell transcriptome sequencing suggests that crustacean cardioactive peptide (CCAP) may be involved. In this study, we cloned the CCAP precursor cDNA (Pj-CCAP) in kuruma shrimp. It was predicted that the gene can be translated into four related peptides and one mature peptide (PFCNAFTGC-NH2), and is highly similar to crustaceans and insects. The predicted Pj-CCAP itself is highly conserved in other crustaceans. RT-qPCR and in situ hybridization analysis showed that Pj-CCAP was most highly expressed in muscle tissue. RNA interference and gene overexpression were used to assess the relationship between Pj-CCAP expression and sand-diving behaviour of shrimp. The results showed that at 6 h post-treatment, the sand-diving rate in the RNA interference group was significantly lower than that in the control group, while the heart rate of shrimp subjected to RNA interference was also significantly, but recovered to baseline levels within 12 h post-treatment. Overexpression resulted in a lower sand diving rate, and a transient increase in heart rate. The results suggest that Pj-CCAP is potentially a negative regulator of sand diving behaviour of kuruma shrimp, and may have an important role in regulating its physiological rhythms and behavioural patterns.
{"title":"Crustacean cardioactive peptide (CCAP) negatively regulates sand-diving behaviour in kuruma shrimp, Penaeus japonicus","authors":"Xinyu Zhou , Jiahan Yu , Huimin Zhang , Yang Zhang , Panpan Wang , Jing Xu , Fei Yu , Jianxin Zhang , Qingqi Zhang , Wazir Ali Baloch , Huan Gao","doi":"10.1016/j.cbpb.2025.111143","DOIUrl":"10.1016/j.cbpb.2025.111143","url":null,"abstract":"<div><div>Kuruma shrimp (<em>Penaeus japonicus</em>) exhibits sand-diving behaviour. The genetic control mechanism of this behaviour is still unclear, although previous single-cell transcriptome sequencing suggests that crustacean cardioactive peptide (CCAP) may be involved. In this study, we cloned the CCAP precursor cDNA (<em>Pj-CCAP</em>) in kuruma shrimp. It was predicted that the gene can be translated into four related peptides and one mature peptide (PFCNAFTGC-NH2), and is highly similar to crustaceans and insects. The predicted <em>Pj-CCAP</em> itself is highly conserved in other crustaceans. RT-qPCR and in situ hybridization analysis showed that <em>Pj-CCAP</em> was most highly expressed in muscle tissue. RNA interference and gene overexpression were used to assess the relationship between <em>Pj-CCAP</em> expression and sand-diving behaviour of shrimp. The results showed that at 6 h post-treatment, the sand-diving rate in the RNA interference group was significantly lower than that in the control group, while the heart rate of shrimp subjected to RNA interference was also significantly, but recovered to baseline levels within 12 h post-treatment. Overexpression resulted in a lower sand diving rate, and a transient increase in heart rate. The results suggest that <em>Pj-CCAP</em> is potentially a negative regulator of sand diving behaviour of kuruma shrimp, and may have an important role in regulating its physiological rhythms and behavioural patterns.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111143"},"PeriodicalIF":1.8,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144857096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-11DOI: 10.1016/j.cbpb.2025.111145
Zhang Shen, Yu Hua, Li Ying
The purpose of this study was to investigate the effects of conjugated linoleic acid (CLA) on fatty acid metabolism through the mammalian target of rapamycin (mTOR) signaling pathway in grass carp (Ctenopharyngodon idella) adipocytes. Grass carp preadipocytes were cultured and treated with various concentrations of CLA (0, 50, 100, 150 and 200 μmol/L). Lipid accumulation was assessed using Oil Red O staining. The regulatory effects on mTOR signaling pathway were evaluated using real-time quantitative PCR (RT-qPCR) and Western Blot (WB). CLA treatment significantly inhibited lipid accumulation and downregulated the expression of genes related to de novo fatty acid synthesis, including fatty acid synthase (fas), acetyl-CoA carboxylase (acc), stearoyl-CoA desaturase 1 (scd1), and sterol regulatory element binding protein-1c (srebp-1c). In contrast, the expression of genes related to fatty acid transport and β-oxidation, such as carnitine palmitoyl transferase-1α (cpt-1α) and hormone-sensitive lipase (hsl), was upregulated. These regulatory effects were found to be concentration-dependent. Co-treatment with the mTOR inhibitor rapamycin further suppressed the mRNA expression of srebp-1c, fas and acc. Furthermore, CLA reduced the protein expression levels of mTOR and nuclear sterol regulatory element-binding protein 1 (nSrebp1), while exerting minimal effect on phosphorylated mTOR (p-mTOR). In summary, CLA appears to modulate lipid metabolism in grass carp adipocytes by inhibiting srebp-1c within the mTOR signaling pathway.
本研究旨在探讨共轭亚油酸（CLA）通过哺乳动物雷帕霉素（mTOR）信号通路对草鱼脂肪细胞脂肪酸代谢的影响。用不同浓度的CLA（0、50、100、150和200 μmol/L）培养草鱼前脂肪细胞。油红O染色检测脂质积累。采用实时荧光定量PCR （RT-qPCR）和Western Blot （WB）方法评价其对mTOR信号通路的调控作用。CLA处理显著抑制脂肪积累，下调脂肪酸合成相关基因的表达，包括脂肪酸合成酶（fas）、乙酰辅酶a羧化酶（acc）、硬脂酰辅酶a去饱和酶1 （scd1）和甾醇调节元件结合蛋白-1c （srebp-1c）。相反，与脂肪酸转运和β-氧化相关的基因，如肉碱棕榈酰转移酶-1α (cpt-1α)和激素敏感脂肪酶（hsl）的表达上调。发现这些调节作用是浓度依赖性的。与mTOR抑制剂雷帕霉素联合治疗进一步抑制srebp-1c、fas和acc的mRNA表达。此外，CLA降低了mTOR和核固醇调控元件结合蛋白1 （nSrebp1）的蛋白表达水平，而对磷酸化mTOR （p-mTOR）的影响很小。综上所述，CLA似乎通过抑制mTOR信号通路中的srebp-1c来调节草鱼脂肪细胞的脂质代谢。
{"title":"Conjugated linoleic acid regulation of fatty acid metabolism by mTOR signaling pathway in grass carp (Ctenopharyngodon idella) adipocytes","authors":"Zhang Shen, Yu Hua, Li Ying","doi":"10.1016/j.cbpb.2025.111145","DOIUrl":"10.1016/j.cbpb.2025.111145","url":null,"abstract":"<div><div>The purpose of this study was to investigate the effects of conjugated linoleic acid (CLA) on fatty acid metabolism through the mammalian target of rapamycin (mTOR) signaling pathway in grass carp (<em>Ctenopharyngodon idella</em>) adipocytes. Grass carp preadipocytes were cultured and treated with various concentrations of CLA (0, 50, 100, 150 and 200 μmol/L). Lipid accumulation was assessed using Oil Red O staining. The regulatory effects on mTOR signaling pathway were evaluated using real-time quantitative PCR (RT-qPCR) and Western Blot (WB). CLA treatment significantly inhibited lipid accumulation and downregulated the expression of genes related to <em>de novo</em> fatty acid synthesis, including fatty acid synthase (<em>fas</em>), acetyl-CoA carboxylase (<em>acc</em>), stearoyl-CoA desaturase 1 (<em>scd1</em>), and sterol regulatory element binding protein-1c (<em>srebp-1c</em>). In contrast, the expression of genes related to fatty acid transport and β-oxidation, such as carnitine palmitoyl transferase-1α (<em>cpt-1α</em>) and hormone-sensitive lipase (<em>hsl</em>), was upregulated. These regulatory effects were found to be concentration-dependent. Co-treatment with the mTOR inhibitor rapamycin further suppressed the mRNA expression of <em>srebp-1c, fas</em> and <em>acc</em>. Furthermore, CLA reduced the protein expression levels of mTOR and nuclear sterol regulatory element-binding protein 1 (nSrebp1), while exerting minimal effect on phosphorylated mTOR (p-mTOR). In summary, CLA appears to modulate lipid metabolism in grass carp adipocytes by inhibiting srebp-1c within the mTOR signaling pathway.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111145"},"PeriodicalIF":1.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144849693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-09DOI: 10.1016/j.cbpb.2025.111144
Lizeth C. Flores-Méndez , Victor Hugo Caña-Bozada , Bruno Gómez-Gil , Crisantema Hernández
High-density aquaculture of Nile tilapia (Oreochromis niloticus) affects welfare by altering cortisol levels, antioxidant response (superoxide dismutase, catalase, and malondialdehyde), and gut microbiota. Agavin is a fructose- and glucose-based polysaccharide containing β-(2-1) and β-(2-6) linkages, known for its potential to mitigate the adverse effects of oxidative stress. This study evaluated the impact of agavin on the intestinal antioxidant response and investigated its correlation with gut microbiota composition and cortisol levels. Fish were fed a control diet (DC0, no agavin) or diets supplemented with 20 g kg−1 (D20) or 40 g kg−1 (D40) of agavin for 110 days. On day 90, all fish were subjected to high-density stress (63 kg m−3). Intestinal superoxide activity, catalase activity, and malondialdehyde concentrations were correlated with previously published plasma cortisol and microbiota data from the same study. Under high-density conditions, the D20 diet increased superoxide dismutase and catalase activity and reduced malondialdehyde concentration compared to the control. Plasma cortisol and malondialdehyde concentration showed a positive correlation (r = 0.67), but plasma cortisol was negatively correlated with superoxide dismutase activity (r = − 0.50). Sixty-nine and 81 operational taxonomic units (OTUs) were significantly correlated with catalase and superoxide dismutase activity, respectively, with families from the Order Cytophagales being strongly positively correlated. Cytophagales was also negatively correlated with plasma cortisol and malondialdehyde concentrations. These results suggest that, under stress conditions, agavin promotes the antioxidant response, modulating the intestinal microbiota and reducing cortisol release, which could indicate its utility as a prebiotic in aquaculture.
尼罗罗非鱼（Oreochromis niloticus）的高密度养殖通过改变皮质醇水平、抗氧化反应（超氧化物歧化酶、过氧化氢酶和丙二醛）和肠道微生物群来影响福利。Agavin是一种以果糖和葡萄糖为基础的多糖，含有β-(2-1)和β-(2-6)键，以其减轻氧化应激不良影响的潜力而闻名。本研究评估了agavin对肠道抗氧化反应的影响，并研究了其与肠道微生物群组成和皮质醇水平的相关性。饲喂对照饲料（DC0，不添加）或在饲料中添加20 g kg-1 （D20）或40 g kg-1 (D40) 110 d。在第90天，所有鱼都受到高密度应激（63 kg m-3）。肠道超氧化物活性、过氧化氢酶活性和丙二醛浓度与先前发表的血浆皮质醇和微生物群数据相关。在高密度条件下，与对照组相比，D20饲粮提高了超氧化物歧化酶和过氧化氢酶活性，降低了丙二醛浓度。血浆皮质醇与丙二醛浓度呈正相关（r = 0.67），与超氧化物歧化酶活性呈负相关（r = - 0.50）。69个操作分类单位（otu）和81个操作分类单位（otu）分别与过氧化氢酶和超氧化物歧化酶活性显著相关，与胞噬目家族呈显著正相关。细胞吞噬体也与血浆皮质醇和丙二醛浓度呈负相关。这些结果表明，在应激条件下，agavin促进抗氧化反应，调节肠道微生物群并减少皮质醇释放，这可能表明其作为水产养殖益生元的用途。
{"title":"The effect of agavin on the intestinal antioxidant response and its relationship to gut microbiota and plasma cortisol in Nile tilapia (Oreochromis niloticus) subjected to overcrowding stress","authors":"Lizeth C. Flores-Méndez , Victor Hugo Caña-Bozada , Bruno Gómez-Gil , Crisantema Hernández","doi":"10.1016/j.cbpb.2025.111144","DOIUrl":"10.1016/j.cbpb.2025.111144","url":null,"abstract":"<div><div>High-density aquaculture of Nile tilapia (<em>Oreochromis niloticus</em>) affects welfare by altering cortisol levels, antioxidant response (superoxide dismutase, catalase, and malondialdehyde), and gut microbiota. Agavin is a fructose- and glucose-based polysaccharide containing β-(2-1) and β-(2-6) linkages, known for its potential to mitigate the adverse effects of oxidative stress. This study evaluated the impact of agavin on the intestinal antioxidant response and investigated its correlation with gut microbiota composition and cortisol levels. Fish were fed a control diet (DC0, no agavin) or diets supplemented with 20 g kg<sup>−1</sup> (D20) or 40 g kg<sup>−1</sup> (D40) of agavin for 110 days. On day 90, all fish were subjected to high-density stress (63 kg m<sup>−3</sup>). Intestinal superoxide activity, catalase activity, and malondialdehyde concentrations were correlated with previously published plasma cortisol and microbiota data from the same study. Under high-density conditions, the D20 diet increased superoxide dismutase and catalase activity and reduced malondialdehyde concentration compared to the control. Plasma cortisol and malondialdehyde concentration showed a positive correlation (<em>r</em> = 0.67), but plasma cortisol was negatively correlated with superoxide dismutase activity (<em>r</em> = − 0.50). Sixty-nine and 81 operational taxonomic units (OTUs) were significantly correlated with catalase and superoxide dismutase activity, respectively, with families from the Order Cytophagales being strongly positively correlated. Cytophagales was also negatively correlated with plasma cortisol and malondialdehyde concentrations. These results suggest that, under stress conditions, agavin promotes the antioxidant response, modulating the intestinal microbiota and reducing cortisol release, which could indicate its utility as a prebiotic in aquaculture.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111144"},"PeriodicalIF":1.8,"publicationDate":"2025-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144823274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The more than 35 described biogenic polyamines have important roles in physiological processes ranging from acid-base buffering to the scavenging of oxygen free radicals. As such they have key cellular- and organismal-level functions in environmental adaptation, cell growth, cell differentiation, fertilization, and biomineralization. To determine cellular polyamine distribution profiles in animals at the base of the phylogenetic tree, the acid-extracted polyamines from cultured cells of a unicellular choanoflagellate and whole bodies of five multicellular invertebrate groups (total 20 species) were quantitatively analyzed using high-performance liquid chromatography and high-performance gas chromatography. Both the choanoflagellate and hydra contained putrescine and spermidine. Diaminopropane, putrescine, cadaverine, norspermidine, spermidine, homospermidine, norspermine, spermine, thermospermine and agmatine were commonly identified among the other invertebrates. In unusual/rare polyamines, aminopropylhomospermidine, homospermine, caldopentamine and homopentamine were found in sponges; aminopropylhomospermidine and canavalmine in the comb jelly; canavalmine, aminopropylcanavalmine and homopyropentamine in jellyfishes; and canavalmine and homopyropentamine in sea anemones. However, long-chain polyamines were not found in soft corals.
{"title":"Polyamine distributions in invertebrates VI: Comparative cellular polyamine analysis of unicellular choanoflagellates and multicellular sponges, comb jellies, hydras, jellyfishes, sea anemones and soft corals","authors":"Koei Hamana , Hidenori Hayashi , Takemitsu Furuchi , Masaru Niitsu","doi":"10.1016/j.cbpb.2025.111141","DOIUrl":"10.1016/j.cbpb.2025.111141","url":null,"abstract":"<div><div>The more than 35 described biogenic polyamines have important roles in physiological processes ranging from acid-base buffering to the scavenging of oxygen free radicals. As such they have key cellular- and organismal-level functions in environmental adaptation, cell growth, cell differentiation, fertilization, and biomineralization. To determine cellular polyamine distribution profiles in animals at the base of the phylogenetic tree, the acid-extracted polyamines from cultured cells of a unicellular choanoflagellate and whole bodies of five multicellular invertebrate groups (total 20 species) were quantitatively analyzed using high-performance liquid chromatography and high-performance gas chromatography. Both the choanoflagellate and hydra contained putrescine and spermidine. Diaminopropane, putrescine, cadaverine, norspermidine, spermidine, homospermidine, norspermine, spermine, thermospermine and agmatine were commonly identified among the other invertebrates. In unusual/rare polyamines, aminopropylhomospermidine, homospermine, caldopentamine and homopentamine were found in sponges; aminopropylhomospermidine and canavalmine in the comb jelly; canavalmine, aminopropylcanavalmine and homopyropentamine in jellyfishes; and canavalmine and homopyropentamine in sea anemones. However, long-chain polyamines were not found in soft corals.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111141"},"PeriodicalIF":1.8,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144796201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-23DOI: 10.1016/j.cbpb.2025.111138
Andrea C. Murillo-Cisneros , Ramón Gaxiola-Robles , Claudia J. Camacho-Hernández , Martha Reyes-Becerril , Orlando Lugo-Lugo , Tania Zenteno-Savín
Developing immortalized cell lines could significantly accelerate studies on marine mammal adaptations to breath-hold diving, ischemia/reperfusion cycles and oxidative stress. In this study, skeletal muscle-derived cells from California sea lions (Zalophus californianus) were transfected with simian virus 40 large T antigen (SV40 LT), a viral oncoprotein known to inactivate cell cycle regulators such as p53 and retinoblastoma (pRB). Although transfection and puromycin selection were successful, transfected cells exhibited morphological abnormalities and reduced viability, suggesting altered cellular proliferation pathways. Significantly higher superoxide dismutase (SOD) and glutathione S-transferase (GST) activities (2- and 6.1-fold, respectively), higher protein oxidative damage (3.9-fold), and lower catalase (CAT) activity (6.9-fold) were observed in transfected cells relative to control (untransfected) cells. These findings suggest that peroxide (H₂O₂) accumulation, likely triggered by genotoxic stress, disrupted cellular proliferation and/or cell death pathways in SV40 LT-transfected skeletal muscle-derived cells from California sea lions. Future studies should consider co-transfection with human telomerase reverse transcriptase (hTERT) and the use of lentiviral delivery systems to enhance transfection efficiency, reduce genotoxic effects, and improve culture stability. This study highlights current challenges and offers potential solutions for generating immortalized marine mammal cell lines.
{"title":"Transfection with SV40 LT promotes oxidative damage in primary cultures of California sea lion muscle cells","authors":"Andrea C. Murillo-Cisneros , Ramón Gaxiola-Robles , Claudia J. Camacho-Hernández , Martha Reyes-Becerril , Orlando Lugo-Lugo , Tania Zenteno-Savín","doi":"10.1016/j.cbpb.2025.111138","DOIUrl":"10.1016/j.cbpb.2025.111138","url":null,"abstract":"<div><div>Developing immortalized cell lines could significantly accelerate studies on marine mammal adaptations to breath-hold diving, ischemia/reperfusion cycles and oxidative stress. In this study, skeletal muscle-derived cells from California sea lions (<em>Zalophus californianus</em>) were transfected with simian virus 40 large T antigen (SV40 LT), a viral oncoprotein known to inactivate cell cycle regulators such as p53 and retinoblastoma (pRB). Although transfection and puromycin selection were successful, transfected cells exhibited morphological abnormalities and reduced viability, suggesting altered cellular proliferation pathways. Significantly higher superoxide dismutase (SOD) and glutathione S-transferase (GST) activities (2- and 6.1-fold, respectively), higher protein oxidative damage (3.9-fold), and lower catalase (CAT) activity (6.9-fold) were observed in transfected cells relative to control (untransfected) cells. These findings suggest that peroxide (H₂O₂) accumulation, likely triggered by genotoxic stress, disrupted cellular proliferation and/or cell death pathways in SV40 LT-transfected skeletal muscle-derived cells from California sea lions. Future studies should consider co-transfection with human telomerase reverse transcriptase (hTERT) and the use of lentiviral delivery systems to enhance transfection efficiency, reduce genotoxic effects, and improve culture stability. This study highlights current challenges and offers potential solutions for generating immortalized marine mammal cell lines.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111138"},"PeriodicalIF":1.9,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144702695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study investigated the behavioral and physiological responses of the freshwater prawn Macrobrachium rosenbergii in relation to fighting activity, aiming to elucidate how energy metabolism varies across locomotory organs under different combat durations. Based on observed fighting times, individuals were divided into control (no fighting), short-duration (16.67 ± 8.22 min), and long-duration (38.33 ± 9.65 min) groups. The movement frequency of the second pereiopods, pleopods, and uropods, along with energy substrates and metabolic enzyme activities in these appendages and abdominal muscles, was analyzed. Results showed that the second pereiopods exhibited the most pronounced increase in beating frequency during prolonged fighting, followed by pleopods and uropods, reflecting their hierarchical involvement in locomotion and aggression. Glycogen content decreased significantly with fighting duration, with the greatest depletion observed in the second pereiopods (up to 36.78 %), compared to moderate decreases in pleopods and abdominal muscles. This indicates a dominant metabolic burden on primary combat appendages. Long-duration fighting induced significant increases in lactate levels, as well as hexokinase (HK) and lactate dehydrogenase (LDH) activities in the second pereiopods and abdominal muscles, suggesting a shift toward anaerobic metabolism. In contrast, short-duration fighting was characterized by elevated malate dehydrogenase (MDH) and cytochrome c oxidase (COX) activities in the second pereiopods and pleopods, reflecting reliance on aerobic pathways. These results demonstrate that M. rosenbergii adopts organ-specific metabolic strategies during aggression. This mechanistic insight lays the groundwork for future aquaculture applications involving behavioral conditioning, dietary interventions, and selective breeding to reduce energy loss and improve animal welfare in intensive farming systems.
{"title":"Metabolic biochemistry of the fighting limbs and muscles of the freshwater prawn, Macrobrachium rosenbergii, reflects agonistic behavior","authors":"Jiangtao Li, Zheng Xiao, Kexuan Zhang, Zhongjie You, Licheng Zeng, Li Lin, Xiaojing Lv","doi":"10.1016/j.cbpb.2025.111137","DOIUrl":"10.1016/j.cbpb.2025.111137","url":null,"abstract":"<div><div>This study investigated the behavioral and physiological responses of the freshwater prawn <em>Macrobrachium rosenbergii</em> in relation to fighting activity, aiming to elucidate how energy metabolism varies across locomotory organs under different combat durations. Based on observed fighting times, individuals were divided into control (no fighting), short-duration (16.67 ± 8.22 min), and long-duration (38.33 ± 9.65 min) groups. The movement frequency of the second pereiopods, pleopods, and uropods, along with energy substrates and metabolic enzyme activities in these appendages and abdominal muscles, was analyzed. Results showed that the second pereiopods exhibited the most pronounced increase in beating frequency during prolonged fighting, followed by pleopods and uropods, reflecting their hierarchical involvement in locomotion and aggression. Glycogen content decreased significantly with fighting duration, with the greatest depletion observed in the second pereiopods (up to 36.78 %), compared to moderate decreases in pleopods and abdominal muscles. This indicates a dominant metabolic burden on primary combat appendages. Long-duration fighting induced significant increases in lactate levels, as well as hexokinase (HK) and lactate dehydrogenase (LDH) activities in the second pereiopods and abdominal muscles, suggesting a shift toward anaerobic metabolism. In contrast, short-duration fighting was characterized by elevated malate dehydrogenase (MDH) and cytochrome <em>c</em> oxidase (COX) activities in the second pereiopods and pleopods, reflecting reliance on aerobic pathways. These results demonstrate that <em>M. rosenbergii</em> adopts organ-specific metabolic strategies during aggression. This mechanistic insight lays the groundwork for future aquaculture applications involving behavioral conditioning, dietary interventions, and selective breeding to reduce energy loss and improve animal welfare in intensive farming systems.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111137"},"PeriodicalIF":1.9,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144711308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The identification of myostatin (mstn) gene functions in salmonids has been complicated by two whole-gene duplications (WGDs), leading to the presence of up to four mstn genes in their genomes, as opposed to the one or two copies found in other vertebrates. In this research, we molecularly cloned and characterized four mstn genes—Sl-mstna1, Sl-mstna2, Sl-mstnb1, and Sl-mstnb2—in the Black Sea trout (Salmo labrax). We propose that Sl-mstnb2 is a pseudogene due to a nonfunctionalization event, while Sl-mstna1, Sl-mstna2, and Sl-mstnb1 are retained in the Salmo labrax genome due to subfunctionalization. Notably, the expression level of the Sl-mstna1 and Sl-mstna2 genes varied across different tissues, implying divergent regulatory mechanisms. Starvation and high temperature increased mRNA expression of Sl-mstna1 and Sl-mstna2, while Sl-mstnb1 transcription was decreased by both stress factors. The tissue-specific transcription and modulation of Sl-mstnb1 were distinctly different from those of the Sl-mstna duplicates, suggesting functional divergence between the Sl-mstna and Sl-mstnb1 genes.
{"title":"Characterization of myostatin genes in Black Sea trout, Salmo labrax, and their differential responses to high temperature and starvation stressors","authors":"Burcu Naz Uzun , Mehtap Bayır , Eyüp Çakmak , Gökhan Arslan , Wenjing Tao , Serpil Turhan , Selma Saoula , Hakan Özkan , Abdulkadir Bayır","doi":"10.1016/j.cbpb.2025.111136","DOIUrl":"10.1016/j.cbpb.2025.111136","url":null,"abstract":"<div><div>The identification of myostatin (<em>mstn</em>) gene functions in salmonids has been complicated by two whole-gene duplications (WGDs), leading to the presence of up to four <em>mstn</em> genes in their genomes, as opposed to the one or two copies found in other vertebrates. In this research, we molecularly cloned and characterized four <em>mstn</em> genes—<em>Sl-mstna1</em>, <em>Sl-mstna2</em>, <em>Sl-mstnb1</em>, and <em>Sl-mstnb2</em>—in the Black Sea trout (<em>Salmo labrax</em>). We propose that <em>Sl-mstnb2</em> is a pseudogene due to a nonfunctionalization event, while <em>Sl-mstna1</em>, <em>Sl-mstna2</em>, and <em>Sl-mstnb1</em> are retained in the <em>Salmo labrax</em> genome due to subfunctionalization. Notably, the expression level of the <em>Sl-mstna1</em> and <em>Sl-mstna2</em> genes varied across different tissues, implying divergent regulatory mechanisms. Starvation and high temperature increased mRNA expression of <em>Sl-mstna1</em> and <em>Sl-mstna2</em>, while <em>Sl-mstnb1</em> transcription was decreased by both stress factors. The tissue-specific transcription and modulation of <em>Sl-mstnb1</em> were distinctly different from those of the <em>Sl-mstna</em> duplicates, suggesting functional divergence between the <em>Sl-mstna</em> and <em>Sl-mstnb1</em> genes.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111136"},"PeriodicalIF":1.9,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144686088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-19DOI: 10.1016/j.cbpb.2025.111129
Francisco A. Leone , Leonardo M. Fabri , Samuel C. Faria , Cintya M. Moraes , Luana G.L. Antonio , Daniel L. Farias , Daniela P. Garçon
Mangrove ecosystems are tidal wetlands with anaerobic, salt-saturated soils that support diverse fauna. Despite intensive studies on the osmoregulatory mechanisms of decapods inhabiting distinct osmotic niches, there is little information on the kinetic characteristics of the gill (Na+, K+)-ATPase of decapod crustaceans inhabiting mangroves. Here, we provide a comprehensive kinetic characterization of the K-phosphatase activity of the gill (Na+, K+)-ATPase in two semi-terrestrial brachyuran crabs, Goniopsis cruentata and Cardisoma guanhumi, which respectively inhabit intermittently flooded lower zones and drier upper zones along the vertical gradient of mangrove environments. This study revealed distinct kinetic profiles between the two species. Goniopsis cruentata shows twice as much K-phosphatase activity and a 3-fold greater affinity for p-nitrophenyl phosphate than Cardisoma guanhumi. Substrate hydrolysis by the Goniopsis cruentata enzyme follows cooperative kinetics, whereas Cardisoma guanhumi displays typical Michaelis-Menten behavior. In both species, stimulation by K+ ions shows cooperative kinetics. Notably, Cardisoma guanhumi also exhibits 25% higher K-phosphatase activity when both K+ and NH₄+ are present. However, whether these differences arise from phylogenetic divergence or reflect physiological adaptation to different mangrove microhabitats remains unresolved. Given the paucity of kinetic studies on (Na+, K+)-ATPase in semi-/terrestrial crabs, our data contribute novel insights into the enzymatic basis of ion transport in decapods inhabiting variable salinity environments.
{"title":"Functional modulation of gill (Na+, K+)-ATPase K-phosphatase activity in mangrove crabs with different degrees of terrestriality","authors":"Francisco A. Leone , Leonardo M. Fabri , Samuel C. Faria , Cintya M. Moraes , Luana G.L. Antonio , Daniel L. Farias , Daniela P. Garçon","doi":"10.1016/j.cbpb.2025.111129","DOIUrl":"10.1016/j.cbpb.2025.111129","url":null,"abstract":"<div><div>Mangrove ecosystems are tidal wetlands with anaerobic, salt-saturated soils that support diverse fauna. Despite intensive studies on the osmoregulatory mechanisms of decapods inhabiting distinct osmotic niches, there is little information on the kinetic characteristics of the gill (Na<sup>+</sup>, K<sup>+</sup>)-ATPase of decapod crustaceans inhabiting mangroves. Here, we provide a comprehensive kinetic characterization of the K-phosphatase activity of the gill (Na<sup>+</sup>, K<sup>+</sup>)-ATPase in two semi-terrestrial brachyuran crabs, <em>Goniopsis cruentata</em> and <em>Cardisoma guanhumi,</em> which respectively inhabit intermittently flooded lower zones and drier upper zones along the vertical gradient of mangrove environments. This study revealed distinct kinetic profiles between the two species. <em>Goniopsis cruentata</em> shows twice as much K-phosphatase activity and a 3-fold greater affinity for <em>p</em>-nitrophenyl phosphate than <em>Cardisoma guanhumi.</em> Substrate hydrolysis by the <em>Goniopsis cruentata</em> enzyme follows cooperative kinetics, whereas <em>Cardisoma guanhumi</em> displays typical Michaelis-Menten behavior. In both species, stimulation by K<sup>+</sup> ions shows cooperative kinetics. Notably, <em>Cardisoma guanhumi</em> also exhibits 25% higher K-phosphatase activity when both K<sup>+</sup> and NH₄<sup>+</sup> are present<em>.</em> However, whether these differences arise from phylogenetic divergence or reflect physiological adaptation to different mangrove microhabitats remains unresolved. Given the paucity of kinetic studies on (Na<sup>+</sup>, K<sup>+</sup>)-ATPase in semi-/terrestrial crabs, our data contribute novel insights into the enzymatic basis of ion transport in decapods inhabiting variable salinity environments.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111129"},"PeriodicalIF":1.9,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144683628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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