Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology最新文献_第5页

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 对过度拥挤应激下尼罗罗非鱼肠道抗氧化反应的影响及其与肠道微生物群和血浆皮质醇的关系

IF 1.8 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-08-09 DOI: 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。

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引用次数: 0

Polyamine distributions in invertebrates VI: Comparative cellular polyamine analysis of unicellular choanoflagellates and multicellular sponges, comb jellies, hydras, jellyfishes, sea anemones and soft corals 多胺在无脊椎动物中的分布VI：单细胞长鞭毛虫和多细胞海绵、栉水母、水螅、水母、海葵和软珊瑚的细胞多胺比较分析。

IF 1.8 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-08-05 DOI: 10.1016/j.cbpb.2025.111141

Koei Hamana , Hidenori Hayashi , Takemitsu Furuchi , Masaru Niitsu

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.

超过35种描述的生物源多胺在生理过程中发挥重要作用，从酸碱缓冲到清除氧自由基。因此，它们在环境适应、细胞生长、细胞分化、受精和生物矿化方面具有关键的细胞和有机体水平的功能。为了确定动物细胞多胺在系统发育树底部的分布特征，我们使用高效液相色谱和高效气相色谱对单细胞鞭毛动物和5个多细胞无脊椎动物群（共20种）的培养细胞中酸提取的多胺进行了定量分析。尾鞭毛虫和水螅均含有腐胺和亚精胺。二氨基丙烷、腐胺、尸胺、去亚精胺、亚精胺、同亚精胺、去精胺、精胺、热精胺和精氨酸在其他无脊椎动物中普遍存在。在异常/稀有多胺中，海绵中检出了氨基丙基同精胺、同精胺、钙多巴胺和同精胺；蜂胶中的氨基丙基同亚精胺和犬avalmine；水母中的鼠尾草碱、氨基丙基鼠尾草碱和同丙戊胺；还有海葵中的鸭avalmine和homopoppentamine。然而，在软珊瑚中并没有发现长链多胺。

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引用次数: 0

Transfection with SV40 LT promotes oxidative damage in primary cultures of California sea lion muscle cells 转染SV40 LT促进加州海狮肌肉细胞原代培养的氧化损伤

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-23 DOI: 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.

开发永生化细胞系可以显著加快海洋哺乳动物对屏气潜水、缺血/再灌注周期和氧化应激的适应性研究。在这项研究中，将加利福尼亚海狮（Zalophus California）的骨骼肌来源细胞转染类人猿病毒40大T抗原（SV40 LT）， SV40 LT是一种已知的病毒癌蛋白，可灭活细胞周期调节因子，如p53和视网膜母细胞瘤（pRB）。虽然转染和嘌呤霉素选择是成功的，但转染后的细胞表现出形态异常和活力降低，表明细胞增殖途径发生了改变。转染细胞的超氧化物歧化酶（SOD）和谷胱甘肽s -转移酶（GST）活性（分别为2-和6.1倍）、蛋白质氧化损伤（3.9倍）和过氧化氢酶（CAT）活性（6.9倍）显著高于对照（未转染）细胞。这些发现表明，过氧化氢（H₂O₂）积累，可能是由基因毒性应激引发的，破坏了来自加州海狮的SV40 rt转染的骨骼肌来源细胞的细胞增殖和/或细胞死亡途径。未来的研究应考虑与人类端粒酶逆转录酶（hTERT）共转染，并使用慢病毒传递系统来提高转染效率，减少基因毒性作用，提高培养稳定性。这项研究强调了当前的挑战，并为产生永生化海洋哺乳动物细胞系提供了潜在的解决方案。

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引用次数: 0

Metabolic biochemistry of the fighting limbs and muscles of the freshwater prawn, Macrobrachium rosenbergii, reflects agonistic behavior 罗氏沼虾（Macrobrachium rosenbergii）战斗肢体和肌肉的代谢生化反应反映了竞争行为

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-23 DOI: 10.1016/j.cbpb.2025.111137

Jiangtao Li, Zheng Xiao, Kexuan Zhang, Zhongjie You, Licheng Zeng, Li Lin, Xiaojing Lv

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.

本研究研究了罗氏沼虾（Macrobrachium rosenbergii）与战斗活动相关的行为和生理反应，旨在阐明不同战斗持续时间下运动器官的能量代谢变化。根据观察到的打斗时间，将个体分为对照组（无打斗）、短时组（16.67±8.22 min）和长时间组（38.33±9.65 min）。分析了第二知觉足类、多足类和尾足类动物的运动频率，以及这些附属物和腹部肌肉的能量底物和代谢酶活性。结果表明，在长时间的打斗中，第二感觉足类动物的殴打频率增加最为明显，其次是pleopod和uro足类动物，这反映了它们在运动和攻击方面的等级参与。糖原含量随着战斗时间的延长而显著下降，第二准足类动物的糖原消耗最多（高达36.78%），而pleopod和腹部肌肉的糖原含量则略有下降。这表明主要战斗附属物的代谢负担占主导地位。长时间的战斗导致第二表足类动物和腹部肌肉中乳酸水平以及己糖激酶（HK）和乳酸脱氢酶（LDH）活性的显著增加，表明向无氧代谢的转变。相比之下，短时间战斗的特征是第二表足类和多足类动物的苹果酸脱氢酶（MDH）和细胞色素c氧化酶（COX）活性升高，反映了对有氧途径的依赖。这些结果表明罗氏芽孢杆菌在攻击过程中采用了器官特异性代谢策略。这一机制见解为未来水产养殖应用奠定了基础，包括行为调节、饮食干预和选择性育种，以减少集约化养殖系统中的能量损失和改善动物福利。

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引用次数: 0

Characterization of myostatin genes in Black Sea trout, Salmo labrax, and their differential responses to high temperature and starvation stressors 黑海鳟鱼肌肉生长抑制素基因的特征及其对高温和饥饿胁迫的差异反应

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-21 DOI: 10.1016/j.cbpb.2025.111136

Burcu Naz Uzun , Mehtap Bayır , Eyüp Çakmak , Gökhan Arslan , Wenjing Tao , Serpil Turhan , Selma Saoula , Hakan Özkan , Abdulkadir Bayır

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.

鲑鱼中肌生长抑制素（mstn）基因功能的鉴定由于两次全基因复制（WGDs）而变得复杂，导致它们的基因组中存在多达四个mstn基因，而在其他脊椎动物中发现的是一个或两个拷贝。本研究在黑海鳟鱼（Salmo labrax）中克隆并鉴定了4个mstn基因sl -mstna1、Sl-mstna2、Sl-mstnb1和sl -mstnb2。我们认为Sl-mstnb2是由于非功能化事件导致的假基因，而Sl-mstna1、Sl-mstna2和Sl-mstnb1由于亚功能化而保留在Salmo labrax基因组中。值得注意的是，Sl-mstna1和Sl-mstna2基因在不同组织中的表达水平存在差异，表明其调控机制存在差异。饥饿和高温使Sl-mstna1和Sl-mstna2的mRNA表达升高，而Sl-mstnb1的转录水平均降低。Sl-mstnb1的组织特异性转录和调控与Sl-mstna重复序列的组织特异性转录和调控明显不同，表明Sl-mstna和Sl-mstnb1基因在功能上存在差异。

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引用次数: 0

Functional modulation of gill (Na+, K+)-ATPase K-phosphatase activity in mangrove crabs with different degrees of terrestriality 不同陆生程度红树林蟹鳃(Na+, K+)-atp酶K-磷酸酶活性的功能调节

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-19 DOI: 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.

红树林生态系统是潮汐湿地，具有厌氧、盐饱和的土壤，支持多种动物。尽管对生活在不同渗透生态位的十足类甲壳类动物的渗透调节机制进行了深入的研究，但对生活在红树林中的十足类甲壳类动物鳃(Na+, K+)- atp酶的动力学特征知之甚少。本文对两种半陆生短爪蟹（Goniopsis cruentata）和冠湖蟹（Cardisoma guanhumi）鳃(Na+, K+)- atp酶的K-磷酸酶活性进行了全面的动力学表征，这两种蟹分别生活在红树林环境垂直梯度上的间歇淹水下带和干旱上带。这项研究揭示了两个物种之间不同的动力学特征。与冠湖田葵相比，小角田葵的k -磷酸酶活性是冠湖田葵的2倍，对对硝基苯磷酸的亲和力是冠湖田葵的3倍。底物的水解是由Goniopsis cruentata酶遵循合作动力学，而冠湖心肌则表现出典型的Michaelis-Menten行为。在这两个物种中，K+离子的刺激表现出合作动力学。值得注意的是，当K+和NH₄+同时存在时，冠湖心病的K-磷酸酶活性也提高了25%。然而，这些差异是由系统发育差异引起的，还是反映了对不同红树林微生境的生理适应，目前尚不清楚。鉴于半/陆生螃蟹(Na+, K+)- atp酶动力学研究的缺乏，我们的数据为生活在可变盐度环境中的十足类动物离子运输的酶基础提供了新的见解。

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引用次数: 0

Metallothionein CgMTIII is involved in zinc binding and accumulation in the Pacific oyster Crassostrea gigas 金属硫蛋白CgMTIII参与了太平洋牡蛎长牡蛎中锌的结合和积累。

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-10 DOI: 10.1016/j.cbpb.2025.111128

Xiang Li , Ning Kong , Wentong Hao , Lixin Feng , Lingling Wang , Linsheng Song

Metallothioneins (MTs) are a family of small, metal-binding proteins that play a pivotal role in metal storage and detoxification. The Pacific oyster Crassostrea gigas is known for its exceptionally high zinc content, and investigating the role of MTs in oyster zinc metabolism will help elucidate the mechanisms underlying its zinc accumulation. In this study, a MT homolog CgMTIII was cloned from C. gigas to investigate its function in zinc binding and accumulation. The transcripts of CgMTIII were distributed in all examined oyster tissues including gill, mantle, adductor muscle, hepatopancreas, gonad, labial palp and haemocytes, with the highest expression in the gill and the lowest in the haemocytes. Immobilized metal ion affinity chromatography (IMAC) demonstrated that the recombinant CgMTIII protein exhibited high retention on the zinc-charged column, confirming its zinc-binding activity. Overexpression of CgMTIII in HEK293T cells led to a 2.34-fold increase in intracellular zinc content after 24 h of exposure to 100 μM zinc, compared to the control group. Moreover, knockdown of CgMTIII through RNA interference resulted in a 24% reduction in zinc content in the gill tissue of oysters. Collectively, CgMTIII exhibited zinc-binding activity and contributed significantly to zinc accumulation in C. gigas. These findings deepen our understanding of the zinc-enrichment mechanism in oysters and provide a theoretical foundation for breeding high‑zinc oyster varieties.

金属硫蛋白（MTs）是一类小的金属结合蛋白，在金属储存和解毒中起关键作用。太平洋牡蛎长牡蛎以其极高的锌含量而闻名，研究MTs在牡蛎锌代谢中的作用将有助于阐明其锌积累的机制。本研究从C. gigas中克隆了一个MT同源基因CgMTIII，以研究其在锌结合和积累中的功能。CgMTIII转录本分布在牡蛎的鳃、套、内收肌、肝胰腺、性腺、唇瓣和血细胞中，其中鳃中表达量最高，血细胞中表达量最低。固定化金属离子亲和层析（IMAC）表明，重组CgMTIII蛋白在带锌柱上具有较高的保留率，证实了其与锌的结合活性。与对照组相比，暴露于100 μM锌24 h后，HEK293T细胞中CgMTIII的过表达导致细胞内锌含量增加2.34倍。此外，通过RNA干扰敲除CgMTIII导致牡蛎鳃组织中锌含量降低24% %。综上所述，CgMTIII表现出锌结合活性，并对C. gigas中锌的积累有显著贡献。这些发现加深了我们对牡蛎锌富集机制的认识，为培育高锌牡蛎品种提供了理论基础。

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引用次数: 0

Developmental exposure to constant elevated temperatures and diel thermal variation alters microRNA expression and performance in zebrafish (Danio rerio). 在斑马鱼的发育过程中，暴露于持续升高的温度和昼夜温差会改变microRNA的表达和表现。

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-09 DOI: 10.1016/j.cbpb.2025.111122

Sana Gavarikar, Paul M. Craig

In their natural environments, fish are exposed to diel thermal fluctuations, the range of which is expected to increase dramatically with climate change. Recently, it has been posited that epigenetic modulators like microRNAs (miRNAs) could buffer fish against such changes. To investigate this, we conducted two sequential experiments using zebrafish (Danio rerio). In Experiment 1, we exposed zebrafish to constant control (CTRL; 28°C), fluctuating (FLUX; 28±5°C), or elevated (HEAT; 33°C) temperatures throughout early development (embryo to juvenile, up to 30 days post-fertilization [dpf]). Throughout early development, we assessed survival, metabolism, and the expression of seven thermosensitive miRNAs and three heat shock proteins (HSPs). While HEAT conditions significantly reduced survival, FLUX exposure did not. Both FLUX and HEAT juveniles exhibited metabolic compensation, though this was more pronounced in FLUX fish. Notably, miR-181a-5p, which regulates mitochondrial biogenesis and respiration, was significantly upregulated in FLUX juveniles, suggesting its involvement in improving metabolic performance. Experiment 2 evaluated the long-term effects of ontogenetic thermal variability. Starting at 30 dpf, remaining fish in both CTRL and FLUX treatments were maintained under CTRL conditions until adulthood. We then assessed miRNA expression in the brain and thermal tolerance (CT_max). miR-181a-5p remained elevated in the brains of adult fish with FLUX developmental histories, though CT_max was unaffected. Together, these findings suggest that early-life exposure to diel thermal fluctuations induces lasting epigenetic changes, potentially optimizing metabolic performance in the short-term at the cost of reduced capacity for further thermal tolerance and stress recovery later in life.

在它们的自然环境中，鱼类面临着剧烈的热波动，其范围预计将随着气候变化而急剧增加。最近，有人认为表观遗传调节剂如microRNAs （miRNAs）可以缓冲鱼类对这些变化的影响。为了研究这一点，我们用斑马鱼（Danio rerio）进行了两次连续实验。在实验1中，我们将斑马鱼暴露于恒定控制(CTRL；28 °C)，波动(FLUX；28 ± 5 °C)，或升高(HEAT；33 °C)的温度贯穿整个早期发育（胚胎到幼崽，直至受精后30 天[dpf]）。在整个早期发育过程中，我们评估了存活、代谢和7种热敏mirna和3种热休克蛋白（HSPs）的表达。虽然HEAT条件显著降低了生存率，但FLUX暴露没有。FLUX和HEAT幼鱼都表现出代谢补偿，尽管这在FLUX鱼中更为明显。值得注意的是，调节线粒体生物发生和呼吸的miR-181a-5p在FLUX幼鱼中显著上调，表明其参与改善代谢表现。实验2评估了个体发生热变率的长期影响。从30 dpf开始，其余的鱼在CTRL和FLUX处理下都保持在CTRL条件下直到成年。然后我们评估了miRNA在脑中的表达和热耐受性（CTmax）。miR-181a-5p在具有FLUX发育历史的成年鱼的大脑中保持升高，但CTmax不受影响。总之，这些发现表明，生命早期暴露于昼夜热波动会引起持久的表观遗传变化，可能会在短期内优化代谢性能，但代价是在生命后期进一步的热耐受性和应激恢复能力降低。

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引用次数: 0

Soluble expression and functional characterization of multi-subunit hemoglobins from the blood clam, Tegillarca granosa 泥蚶多亚基血红蛋白的可溶性表达及功能表征

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-09 DOI: 10.1016/j.cbpb.2025.111125

Weifeng Zhang , Tiancheng Chen , Yazhuo Hu , Xuejia Li , Xizhi Shi , Yongbo Bao

The Escherichia coli expression system, combined with site-directed mutagenesis, is a powerful tool for investigating the structure-function relationship of hemoglobin (Hb). However, producing biologically active recombinant hemoglobin (rHb) from invertebrates, especially in multimeric forms, remains difficult. Here, we describe the recombinant expression, purification, and characterization of homodimeric rHbI and heterotetrameric rHbII from the blood clam Tegillarca granosa. Optimization of expression conditions demonstrated that high yields (65.4–135.6 g/L) of soluble T. granosa rHbs could be achieved using the JM109(DE3) strain with 15 °C induction, 0.25 mM isopropyl β-D-thiogalactoside (IPTG) levels, and codon optimization. Circular dichroism and ultraviolet-visible spectroscopy verified that purified rHbs exhibited a secondary structure similar to that of native Hbs (nHbs) and preserved their diverse ligand-binding capacities. Oxygen-binding assays further showed that the oxygen affinity of rHbs was comparable to that of nHbs. Nitrite reduction assays revealed that T. granosa nHbs exhibited weak nitrite reductase activity (0.049–0.116 M⁻¹ s⁻¹ at 25 °C), generally lower than that of vertebrate globins. However, rHbs, particularly rHbI, displayed substantially higher reduction rate constants (0.147–4.589 M⁻¹ s⁻¹ at 25 °C) than nHbs, possibly due to differences in structural conformation or the N-terminal tag carried by rHbs. Overall, the recombinant expression and purification method established in this study provides a valuable framework for future research on the structure-function relationships of T. granosa Hb and invertebrate Hbs more broadly.

大肠杆菌表达系统，结合定点诱变，是研究血红蛋白（Hb）结构-功能关系的有力工具。然而，从无脊椎动物中生产具有生物活性的重组血红蛋白（rHb），特别是以多聚体形式，仍然很困难。在这里，我们描述了从泥蚶中重组表达、纯化和表征同二聚体和异四聚体的rHbII。优化表达条件表明，JM109（DE3）菌株在15°C诱导、0.25 mM异丙基β- d -硫代半乳糖苷（IPTG）水平和密码子优化条件下，可获得高产量（65.4-135.6 g/L）的可溶性大叶黄菌。圆二色性和紫外可见光谱验证了纯化的rHbs具有与天然Hbs （nHbs）相似的二级结构，并保留了其不同的配体结合能力。氧结合实验进一步表明，rHbs的氧亲和力与nHbs相当。亚硝酸盐还原实验表明，T. granosa nHbs具有较弱的亚硝酸盐还原酶活性（25°C时为0.049 ~ 0.116 M−1 s−1），普遍低于脊椎动物珠蛋白。然而，rHbs，特别是rHbI，表现出明显高于nHbs的还原速率常数（25°C时为0.147-4.589 M−1 s−1），这可能是由于rHbs结构构象的差异或其携带的n端标签。综上所述，本研究建立的重组表达和纯化方法为今后更广泛地研究巨噬菌Hb和无脊椎动物Hb的结构-功能关系提供了有价值的框架。

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引用次数: 0

Effects of gonadorelin on gonadotropin expression, plasma sex steroid concentrations and ovarian follicle dynamics in mature tambaqui (Colossoma macropomum) 促性腺激素对成熟柽柽鱼促性腺激素表达、血浆性类固醇浓度及卵泡动力学的影响

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-09 DOI: 10.1016/j.cbpb.2025.111126

Rosilane Gomes de Souza Oliveira , Iraní Silva de Morais , Rômulo Veiga Paixão , Izabel Correa Bandeira , Wallice Luiz Paxiúba Duncan , Fernanda Loureiro de Almeida O'Sullivan

In tambaqui (Colossoma macropomum), a rheophilic Amazonian fish of significant aquaculture importance that typically fails to spawn under captive conditions, gonadorelin has demonstrated potential in inducing ovulation and enabling strip spawning. However, the molecular mechanisms underlying gonadorelin action, particularly on the transcriptional regulation of follicle-stimulating hormone (fshβ) and luteinizing hormone (lhβ), remain poorly understood and warrant further investigation. The present study aimed to investigate the efficacy of this synthetic drug in modulating the expression of gonadotropins and in enhancing circulating concentrations of 17β-estradiol (E2), 17α-hydroxyprogesterone (17α-OHP), and testosterone (T) in tambaqui; as well as its effects on the developmental dynamics and maturation stages of ovarian follicles. Gonadorelin at 60 μg kg⁻¹ body weight had no statistical effect on lhβ expression, but decreased fshβ expression in females (p = 0.021). E₂, 17α-OHP and T increased in all treated fish. In the treated group, the appearance of post-ovulatory complexes and the presence of eggs in the coelomic cavity indicated that 6 of 9 (67 %) of the females ovulated, while none of the control group did. The findings indicate that gonadorelin suppresses fshβ transcription in mature female tambaqui, and this downregulation may promote final oocyte maturation and ovulation by favoring a hormonal environment with increased progesterone influence. Hence, our data provides the first detailed analysis of the effects of a drug on tambaqui gonadotropins and sex steroids and sets a foundation for refining dose strategies to improve large-scale controlled breeding practices in tambaqui aquaculture.

在tambaqui （Colossoma macropomum）中，一种具有重要水产养殖意义的亚马逊嗜流鱼，通常在圈养条件下无法产卵，促性腺激素已被证明具有诱导排卵和使条形产卵的潜力。然而，促性腺激素作用的分子机制，特别是促卵泡激素（fshβ）和黄体生成素（lhβ）的转录调节，仍然知之甚少，需要进一步研究。本研究旨在探讨该合成药物对调节促性腺激素的表达和提高血中17β-雌二醇（E2）、17α-羟孕酮（17α-OHP）和睾酮(T)浓度的作用；以及其对卵巢卵泡发育动态和成熟阶段的影响。60 μg kg−1体重的促性腺激素对雌性体内lhβ表达无显著影响，但可降低fshβ表达（p = 0.021）。E2、17α-OHP和T均升高。在治疗组中，排卵后复合体的出现和体腔中卵子的存在表明，9只雌性中有6只（67%）排卵，而对照组没有排卵。研究结果表明，促性腺激素抑制成熟雌性坦巴基鱼的fshβ转录，这种下调可能通过增加孕酮影响的激素环境促进最终卵母细胞成熟和排卵。因此，我们的数据首次详细分析了一种药物对坦巴基鱼促性腺激素和性类固醇的影响，并为完善剂量策略以改善坦巴基鱼养殖的大规模控制养殖实践奠定了基础。

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引用次数: 0