Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology最新文献_第6页

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

Effects of dietary corn silk (Zea mays L.) on growth, immune and antioxidant pathways, histological morphology, gut microbiome, and sensitivity to acute ammonia exposure in the koi carp (Cyprinus carpio var. koi) 饲粮添加玉米丝对锦鲤生长、免疫和抗氧化途径、组织学形态、肠道微生物组以及急性氨暴露敏感性的影响

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-07 DOI: 10.1016/j.cbpb.2025.111127

Nguyen Vu Linh , Luu Tang Phuc Khang , Nguyen Dinh-Hung , Suwanna Wisetkaeo , Phatthanaphong Therdtatha , Papungkorn Sangsawad , Supreya Wannavijit , Jakree Jitjumnong , Patima Permpoonpattana , Hien Van Doan

Corn silk is a promising aquafeed additive due to its antioxidant, antibacterial, and anti-inflammatory properties. This study evaluated the impact of graded dietary levels of powdered corn silk (PCS) on growth, immunity, antioxidant enzyme activities, histology, gut microbiota composition, gene expression profiles, and sensitivity to acute ammonia exposure in koi carp (Cyprinus carpio var. koi). A total of 300 fish were assigned to five dietary treatments (0, 5, 10, 20, and 30 g/kg PCS) for eight weeks. Fish fed the PCS-30 diet showed significant improvements in growth parameters, including weight gain (WG) and specific growth rate (SGR) (p < 0.05). Antioxidant capacity (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid); ABTS) and superoxide dismutase (SOD) activity, increased dose-dependently, while malondialdehyde (MDA) levels remained unchanged (p > 0.05). Expression of the immune and antioxidant genes interleukin 8 (il-8), lysozyme (lyz), catalase (cat), nuclear factor-κB (Nf-κB), and toll-like receptor 4 (tlr4) was significantly upregulated in the PCS-20 and PCS-30 groups (p < 0.05). Histological analysis revealed progressive improvements in anterior intestinal morphology, with the PCS-30 group exhibiting the greatest villus height and width. Gut microbiota analysis showed increased relative abundance of beneficial taxa (Gammaproteobacteria, Lactobacillales, Prevotella), which positively correlated with growth and antioxidant markers, and negatively with MDA levels. Survival under acute ammonia exposure was highest in the PCS-20 and PCS-30 groups. These findings demonstrate the efficacy of PCS, particularly at 30 g/kg, in enhancing growth, immunity, and stress resilience.

玉米丝具有抗氧化、抗菌和抗炎的特性，是一种很有前途的水产饲料添加剂。本试验研究了不同饲料水平玉米丝粉（PCS）对锦鲤生长、免疫、抗氧化酶活性、组织学、肠道菌群组成、基因表达谱和急性氨暴露敏感性的影响。将300尾鱼分为5种饲料处理（0、5、10、20和30 g/kg PCS），为期8周。饲喂PCS-30饲料的鱼在增重（WG）和特定生长率（SGR）等生长参数方面均有显著提高（p  0.05）。免疫和抗氧化基因白介素8 （il-8）、溶菌酶（lyz）、过氧化氢酶（cat）、核因子-κB （Nf-κB）和toll样受体4 （tlr4）的表达在PCS-20和PCS-30组中显著上调(p

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引用次数: 0

Microplastic and Vibrio harveyi co-exposure induces oxidative stress in big-belly seahorse Hippocampus abdominalis 微塑料与哈维弧菌共暴露诱导大腹海马海马腹肌氧化应激

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-05 DOI: 10.1016/j.cbpb.2025.111124

Jin A Kim , Jong Yoon Lee , Seong Don Hwang , Jun-Hwan Kim , Cheol Young Choi

Microplastics in marine environments can act as vectors for pathogenic bacteria, increasing the risk of infection and physiological stress in marine organisms. Seahorses, which inhabit coastal ecosystems, may be vulnerable to the combined effects of microplastics and bacterial pathogens such as Vibrio harveyi. Therefore, the aim of this study was to investigate the effect of microplastic and bacteria co-exposure on big-belly seahorse (Hippocampus abdominalis). Microbeads (MBs) were introduced via water exposure, while V. harveyi was administered by injection. Seahorses were exposed to microbeads (MBs; concentration; 50 beads/L; size, 0.2 and 1.0 μm), V. harveyi (1 × 10³ CFU/mL), and a combination of both for 4 days to evaluate oxidative stress responses. Tissue sampling was performed at 0, 1, 2, and 4 days. Notably, MB accumulation in the gills and intestine was significantly higher in the group co-exposured to MB and bacteria than in the single exposure groups. V. harveyi copy number in the liver was significantly higher in the co-exposure groups, especially in the Vh + MB1.0 group. Compared with the control group, MB and V. harveyi co-exposure significantly upregulated antioxidant response, including superoxide dismutase, catalase, and 1-cysteine peroxiredoxin mRNA and protein levels. Additionally, hydrogen peroxide levels and DNA damage were elevated in the Vh + MB1.0 group, indicating enhanced oxidative stress and genotoxicity. In conclusion, MB and V. harveyi co-exposure induces oxidative stress and DNA damage in seahorse, suggesting potential ecological risks associated with simultaneous exposure to microplastics and marine pathogens in coastal ecosystems.

海洋环境中的微塑料可作为致病菌的载体，增加了海洋生物感染和生理应激的风险。生活在沿海生态系统中的海马可能很容易受到微塑料和哈维弧菌等细菌病原体的综合影响。因此，本研究旨在探讨微塑料和细菌共同暴露对大腹海马（Hippocampus abdominalis）的影响。微珠（MBs）通过水暴露引入，而哈维氏弧菌通过注射给药。海马暴露于微珠（MBs）；浓度;50个珠子/ L;大小，0.2和1.0 μm)， V. harveyi (1 × 103 CFU/mL)，以及两者的组合4天，以评估氧化应激反应。在第0、1、2和4天进行组织取样。值得注意的是，与单独暴露组相比，MB和细菌共同暴露组的鳃和肠道中的MB积累量显著增加。共暴露组肝脏中V. harveyi拷贝数显著升高，特别是在Vh + MB1.0组。与对照组相比，MB和V. harveyi共暴露显著上调抗氧化反应，包括超氧化物歧化酶、过氧化氢酶和1-半胱氨酸过氧化物还蛋白mRNA和蛋白水平。此外，过氧化氢水平和DNA损伤在Vh + MB1.0组升高，表明氧化应激和遗传毒性增强。综上所述，MB和V. harveyi共暴露可诱导海马氧化应激和DNA损伤，提示沿海生态系统中同时暴露于微塑料和海洋病原体可能存在生态风险。

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引用次数: 0

Cloning of the Na+-K+-2Cl−(NKCC) transporter in Macrobrachium rosenbergii and its role in osmoregulation 罗氏沼虾Na+-K+-2Cl - (NKCC)转运体的克隆及其在渗透调节中的作用

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-07-03 DOI: 10.1016/j.cbpb.2025.111123

Weifeng Gao, Lu Chen, Qianqian Ding, Xiaojian Gao, Yao Zhang, Xiaojun Zhang, Qun Jiang

The full-length cDNA sequences of two Na-K-2Cl cotransporter (NKCC) isoforms, MrNKCC1 and MrNKCC2, were identified in Macrobrachium rosenbergii. MrNKCC1 contains a 3183 bp open reading frame (ORF) encoding 1060 amino acids, while MrNKCC2 has a 2679 bp ORF encoding 892 amino acids. Both isoforms share a similar predicted secondary structure, featuring 12 transmembrane domains and a C-terminal, and their amino acid sequences and domain arrangement are highly conserved. Phylogenetic analysis groups fish and crustacean NKCC isoforms into distinct branches, each containing two isoforms. Expression analysis using real-time quantitative PCR (qPCR) revealed distinct tissue distribution patterns. MrNKCC1 was broadly expressed, with the highest levels in the intestine, whereas MrNKCC2 was predominantly found in the gill, hepatopancreas, and epidermis, with peak expression in the gill. To examine their roles in osmoregulation, prawns were exposed to 24 ‰ seawater for 96 h. Hemolymph concentrations of Cl⁻, K⁺, and Na⁺ increased, peaking at 24, 24, and 12 h, respectively. During this period, MrNKCC1 expression was suppressed, while MrNKCC2 expression significantly increased at 2 h after seawater exposure and then returned to initial levels. Gill filaments showed significant histological changes under seawater exposure, including the expansion and proliferation of chloride-secreting cells. Following a 96-h seawater exposure, prawns were transferred to freshwater. This transition led to a decline in plasma Cl⁻, K⁺, and Na⁺ concentrations, accompanied by a marked increase in MrNKCC1 and MrNKCC2 expression. The rapid transcriptional response of both MrNKCC isoforms to salinity fluctuations highlights their critical role in the osmoregulatory mechanisms of M. rosenbergii.

在罗氏沼虾（Macrobrachium rosenbergii）中鉴定了两个Na-K-2Cl共转运体（NKCC）亚型MrNKCC1和MrNKCC2的全长cDNA序列。MrNKCC1含有3183 bp的开放阅读框（ORF），编码1060个氨基酸，MrNKCC2含有2679 bp的ORF，编码892个氨基酸。两种异构体具有相似的预测二级结构，均具有12个跨膜结构域和一个c -末端，其氨基酸序列和结构域排列高度保守。系统发育分析将鱼类和甲壳类的NKCC亚型分为不同的分支，每个分支包含两个亚型。实时定量PCR （qPCR）表达分析揭示了不同的组织分布模式。MrNKCC1广泛表达，在肠道中表达量最高，而MrNKCC2主要存在于鳃、肝胰腺和表皮中，在鳃中表达量最高。为了研究它们在渗透调节中的作用，将对虾暴露在24‰海水中96 h，血淋巴中Cl−、K+和Na+浓度分别在24、24和12 h达到峰值。在此期间，MrNKCC1的表达受到抑制，而MrNKCC2的表达在海水暴露后2 h显著增加，然后恢复到初始水平。在海水照射下，鳃丝发生了明显的组织学变化，包括分泌氯化物的细胞的扩张和增殖。在海水中暴露96小时后，将对虾转移到淡水中。这种转变导致血浆Cl−、K+和Na+浓度下降，同时MrNKCC1和MrNKCC2表达显著增加。两种MrNKCC亚型对盐度波动的快速转录反应突出了它们在罗氏m.r obergii渗透调节机制中的关键作用。

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引用次数: 0

Comparative sensitivity of marine invertebrate lipids to oxidation under prooxidant conditions in vitro 海洋无脊椎动物脂类在体外促氧化条件下对氧化的比较敏感性。

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-06-26 DOI: 10.1016/j.cbpb.2025.111121

Aleksandra Istomina, Viktor Chelomin, Olesya Elovskaya, Nadezhda Dovzhenko, Valentina Slobodskova, Lyudmila Kolosova, Yuliya Fedorets

The current study compared the degree of oxidative damage induced by Fenton reagents (ferrous sulfate and hydrogen peroxide) in the digestive tissue homogenates of 13 species of marine invertebrates (mollusks and echinoderms). Lipid peroxidation (thiobarbituric acid reactive substances, TBARS) was used to estimate the potential resistance to lipid peroxidation in the studied organisms. These data showed that invertebrates can be divided into two groups. The first group included sea urchins and gastropod mollusks, which had a high level of integrated antiradical activity (IAA) and a low level of lipids available for peroxidation (LPA). The second group included sea stars and bivalves, which have low levels of IAA and varying amounts of LPA. On the basis of these data, it is hypothesized that species occupying the upper littoral and intertidal zones (sea urchins, gastropod mollusks), and which are exposed to fluctuations in factors that affect metabolic processes and thus alter the levels of reactive oxygen species, are adapted for such settings through a high antioxidant potential. Conversely, species that inhabit deeper waters under more stable conditions (bivalves, sea stars), do not need to maintain a high concentration of low-molecular-weight antioxidants. This may represent an important adaptation of these species to environmental fluctuations in their natural habitats.

本研究比较了Fenton试剂（硫酸亚铁和过氧化氢）对13种海洋无脊椎动物（软体动物和棘皮动物）消化组织匀浆的氧化损伤程度。脂质过氧化（硫代巴比妥酸反应物质，TBARS）被用来评估研究生物对脂质过氧化的潜在抗性。这些数据表明，无脊椎动物可以分为两类。第一组包括海胆和腹足类软体动物，它们具有高水平的综合抗自由基活性（IAA）和低水平的过氧化脂质（LPA）。第二组包括海星和双壳类动物，它们的IAA含量低，LPA含量不同。在这些数据的基础上，假设占据上滨海和潮间带的物种（海胆、腹足类软体动物），暴露于影响代谢过程的因素波动中，从而改变活性氧的水平，通过高抗氧化潜力适应这种环境。相反，在更稳定的条件下生活在更深水域的物种（双壳类，海星）不需要维持高浓度的低分子量抗氧化剂。这可能代表了这些物种对其自然栖息地环境波动的重要适应。

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引用次数: 0

Iron mineral formation using chiton-derived ferritins 利用石壳衍生铁蛋白形成铁矿物。

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-06-20 DOI: 10.1016/j.cbpb.2025.111120

Shenghao Du, Juanli Liao, Dawei Sun, Chuang Liu

The teeth of chitons, a group of marine mollusks, rank among the hardest biomaterials found on earth. These teeth are composed of structurally ordered iron oxide minerals. Ferritin is an iron-storage protein known to regulate iron metabolism across diverse organisms; however, its role in iron biomineralization in chiton teeth remains unclear. Herein, ferritins were isolated from chiton hemolymph. Transcriptome- and proteome-based analysis revealed that the main ferritins in chiton hemolymph are also found in chiton radulae, which is supported by their elevated transcriptional abundance in radular tissues. In vitro assays revealed that these ferritins can bind iron ions and promote the formation of iron-based minerals. Ex vivo experiments using unmineralized radular scaffolds further showed that chiton ferritins facilitate iron oxide biomineralization, although the resulting crystals differed structurally from the native magnetite found in mature teeth. Phylogenetic analysis confirmed the conservation of ferritins among chitons, indicative of their universal role in tooth formation in chitons. Our findings advance the molecular understanding of iron-based biomineralization in chitons and provide insights for the design of synthetic iron-based nanomaterials.

石鳖是一种海洋软体动物，它们的牙齿是地球上发现的最坚硬的生物材料之一。这些牙齿是由结构有序的氧化铁矿物组成的。铁蛋白是一种铁储存蛋白，可以调节不同生物体的铁代谢；然而，其在石鳖牙齿铁生物矿化中的作用尚不清楚。本文从石鳖血淋巴中分离出铁蛋白。基于转录组学和蛋白质组学的分析表明，石鳖血淋巴中的主要铁蛋白也存在于石鳖结节组织中，这与它们在结节组织中的转录丰度升高有关。体外实验表明，这些铁蛋白可以结合铁离子，促进铁基矿物的形成。使用未矿化的根状支架进行的体外实验进一步表明，石壳铁蛋白促进氧化铁生物矿化，尽管所产生的晶体在结构上与成熟牙齿中发现的天然磁铁矿不同。系统发育分析证实了铁蛋白在石鳖中的保守性，表明铁蛋白在石鳖牙齿形成中的普遍作用。我们的发现促进了对石粒中铁基生物矿化的分子理解，并为合成铁基纳米材料的设计提供了见解。

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引用次数: 0

Effects on life parameters and fatty acid expression profiles in response to elevated temperatures in CRISPR/Cas9-mediated gene–targeted mutants of two desaturase (Δ9–3 and Δ5/6) genes in the water flea Daphnia magna 大水蚤（Daphnia magna）两种去饱和酶（Δ9-3和Δ5/6）基因CRISPR/ cas9介导的基因靶向突变体在高温下对生命参数和脂肪酸表达谱的影响

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-06-18 DOI: 10.1016/j.cbpb.2025.111119

Deok-Seo Yoon , Eunjin Byeon , Seong Chan Yun , Haksoo Jeong , Jin-Sol Lee , Yunfei Sun , Eun-Ji Won , Heum Gi Park , Zhou Yang , Atsushi Hagiwara , Min-Chul Lee , Jae-Seong Lee

Fatty-acid desaturation is central to membrane homeostasis and thermal performance in ectotherms. We disrupted the Δ9–3 (MUFA-specific) and Δ5/6 (long-chain PUFA-specific) desaturase genes in Daphnia magna with CRISPR/Cas9 and compared wild-type and knockout lines at 23 °C and 28 °C. Loss of Δ9–3 substantially depleted monounsaturated fatty acids, especially oleic and palmitoleic acids, with a compensatory rise in total polyunsaturates. These lipid shifts coincided with enhanced growth and fecundity under benign temperature but translated into reduced reproductive output when heat stress was imposed, indicating a context-dependent trade-off. In contrast, deletion of Δ5/6 selectively impaired the n-6 pathway at high temperature, leading to pronounced reductions in total PUFA reserves and a severe decline in offspring production; the usual growth benefit of warming was also abolished. Together, the data reveal that Δ9–3 supports MUFA supply needed for baseline metabolism, whereas Δ5/6 maintains PUFA reserves critical for reproduction under thermal challenge. Divergent desaturase functions thus play distinct roles in lipid remodeling and contribute to climate resilience in freshwater zooplankton.

脂肪酸去饱和对恒温动物的膜稳态和热性能至关重要。我们用CRISPR/Cas9破坏了大水蚤（Daphnia magna）的Δ9-3 （mufa特异性）和Δ5/6（长链pufa特异性）去饱和酶基因，并在23 °C和28 °C下比较了野生型和敲除系。Δ9-3大量消耗的单不饱和脂肪酸，特别是油酸和棕榈油酸，随着总多不饱和脂肪酸的补偿性增加而减少。这些脂质变化与良性温度下的生长和繁殖力增强相一致，但在施加热应激时转化为生殖产量降低，表明了环境依赖的权衡。相反，Δ5/6的缺失在高温下选择性地破坏了n-6通路，导致PUFA总储备明显减少，后代产量严重下降；气候变暖通常带来的增长效益也被取消了。总之，数据显示Δ9-3支持基线代谢所需的多聚脂肪酸供应，而Δ5/6维持在热挑战下繁殖所需的多聚脂肪酸储备。因此，不同的去饱和酶功能在淡水浮游动物的脂质重塑和气候适应中发挥着不同的作用。

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引用次数: 0

Molecular characterization of HSP70, HSP90a, expression responses and biochemical changes in yellowfin seabream Acanthopagrus latus (Houttuyn 1782) under temperature stress HSP70、HSP90a在温度胁迫下的分子特征、表达响应及生化变化

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology

Pub Date : 2025-06-17 DOI: 10.1016/j.cbpb.2025.111118

Jing-Jing Wu , Ya-Ling Li , Jin-Min Pan , Hua-Yang Guo , Bao-Suo Liu , Nan Zhang , Lin Xian , Ke-Cheng Zhu , Dian-Chang Zhang

This study investigates the molecular characteristics and expression profiles of heat shock proteins (HSP70, HSP90a, and HSP90a1) and biochemical changes in yellowfin seabream (Acanthopagrus latus) under acute temperature stress. The full-length cDNA sequences of these HSPs were cloned and analyzed, revealing open reading frames encoding 639, 730, and 724 amino acids, respectively, each containing conserved motifs indicative of their roles in the stress response. Tissue-specific expression analysis showed that AlHSP70 was predominantly expressed in the kidney, while AlHSP90a and AlHSP90a1 exhibited highest expression levels in the heart of healthy adult males. Under acute temperature stress, quantitative real-time PCR (qPCR) demonstrated that hepatic expression levels of these genes in juvenile fish initially increased, followed by a decrease and stabilization over time. Concurrently, activities of antioxidant enzymes—total superoxide dismutase (T-SOD) and catalase (CAT)—and the metabolic enzyme lactate dehydrogenase (LDH) in the liver showed a similar pattern of initial elevation followed by decline. These findings suggest that HSPs play a critical role in the physiological response to temperature-induced stress, contributing to protection against oxidative damage in A. latus. This study provides fundamental insights into the molecular mechanisms underlying thermal stress acclimation in this species.

本研究探讨了急性温度胁迫下黄鳍鲷（Acanthopagrus latus）热休克蛋白（HSP70、HSP90a和HSP90a1）的分子特征、表达谱及生化变化。克隆并分析了这些热休克蛋白的全长cDNA序列，揭示了分别编码639、730和724个氨基酸的开放阅读框，每个阅读框都含有保守的基序，表明它们在应激反应中的作用。组织特异性表达分析显示，AlHSP70主要在肾脏中表达，而AlHSP90a和AlHSP90a1在健康成年男性的心脏中表达水平最高。在急性温度胁迫下，实时荧光定量PCR （qPCR）结果显示，这些基因在幼鱼肝脏中的表达水平最初升高，随后随着时间的推移逐渐下降并趋于稳定。同时，肝脏中抗氧化酶——总超氧化物歧化酶（T-SOD）和过氧化氢酶（CAT）以及代谢酶乳酸脱氢酶（LDH）的活性也呈现出类似的先升高后下降的模式。这些发现表明，热休克蛋白在温度诱导应激的生理反应中起关键作用，有助于保护拉胡斯免受氧化损伤。本研究为该物种热胁迫驯化的分子机制提供了基本的见解。

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引用次数: 0