A Sulfated Polysaccharide from Saccharina japonica Suppresses LPS-Induced Inflammation Both in a Macrophage Cell Model via Blocking MAPK/NF-κB Signal Pathways In Vitro and a Zebrafish Model of Embryos and Larvae In Vivo

Inflammation is a complicated host-protective response to stimuli and toxic conditions, and is considered as a double-edged sword. A sulfated Saccharina japonica polysaccharide (LJPS) with a sulfate content of 9.07% showed significant inhibitory effects against lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells and zebrafish. Its chemical and structural properties were investigated via HPLC, GC, FTIR, and NMR spectroscopy. In vitro experiments demonstrated that LJPS significantly inhibited the generation of nitric oxide (NO) and prostaglandin E₂ (PGE₂) via the downregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and suppressed pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1β production via the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signal pathways in LPS-induced RAW 264.7 cells. Moreover, LJPS showed strong protective effects against LPS-induced inflammatory responses in zebrafish, increasing the survival rate, reducing the heart rate and yolk sac edema size, and inhibiting cell death and the production of intracellular reactive oxygen species (ROS) and NO. Its convenience for large-scale production and significant anti-inflammatory activity indicated the potential application of LJPS in functional foods, cosmetics, and pharmaceutical industries.     

Using fish models to investigate the links between microbiome and social behaviour: The next step for translational microbiome research?

Recent research has revealed surprisingly important connections between animals’ microbiome and social behaviour. Social interactions can affect the composition and function of the microbiome; conversely, the microbiome affects social communication by influencing the hosts’ central nervous system and peripheral chemical communication. These discoveries set the stage for novel research focusing on the evolution and physiology of animal social behaviour in relation to microbial transmission strategies. Here, we discuss the emerging roles of teleost fish models and their potential for advancing research fields, linked to sociality and microbial regulation. We argue that fish models, such as the zebrafish (Danio rerio, Cyprinidae), sticklebacks (?Gasterosteidae), guppies (Poeciliidae) and cleaner–client dyads (e.g., obligate cleaner fish from the Labridae and Gobiidae families and their visiting clientele), will provide valuable insights into the roles of microbiome in shaping social behaviour and vice versa, while also being of direct relevance to the food and ornamental fish trades. The diversity of fish behaviour warrants more interdisciplinary research, including microbiome studies, which should have a strong ecological (field‐derived) approach, together with laboratory‐based cognitive and neurobiological experimentation. The implications of such integrated approaches may be of translational relevance, opening new avenues for future investigation using fish models.     

Aluminium chloride‐induced toxicity in zebrafish larvae

Embryos at shield stage and larvae at protruding mouth stage were exposed to different concentrations of aluminium chloride (AlCl₃) for 72 h with the purpose to analyse their phenotype and lethality. After 24, 48 and 72 h of treatment, higher toxicity of the metal was observed on larvae with minimal lethal concentration of 0.25, 0.20 and 0.08 mm , respectively, while for embryos the corresponding values were 40, 25 and 16 mm . We observed pericardial oedema and alteration of heart rate in 50% of larvae after 48 h of exposure to 100 μm . In larvae exposed to the same concentration, there was also a neurological injury at the level of glial cells, with the number of glial fibrillary acidic protein‐positive cells being significantly reduced. This study confirms the toxic nature of this metal and shows that aluminium could also interestingly represent a cardiotoxin in addition to its neurotoxic ability.     

斑马鱼幼鱼嗜中性粒细胞对副溶血弧菌清除的动态变化

副溶血弧菌(Vibrio parahaemolyticus)是人-兽-鱼共患的致病菌,严重威胁食品安全和人类的健康。为了研究宿主嗜中性粒细胞清除细菌感染的动态变化过程,利用红色荧光蛋白标记的副溶血弧菌Vp57~(RFP)菌株感染受精后48 h(hours post fertilization,hpf)的斑马鱼(Danio rerio)幼鱼耳泡,建立局部感染模型,并以大肠杆菌(Escherichia coli)Ec01菌株为对照,系统比较了副溶血弧菌和大肠杆菌的半数致死剂量(median lethal dose,LD_(50))、存活曲线以及和嗜中性粒细胞相互作用的动态过程,RT-qPCR验证炎症相关基因il1b和il10在感染前后的表达量变化。结果显示副溶血弧菌和大肠杆菌感染斑马鱼幼鱼的LD_(50)分别为7.90×10~8 CFU/mL和1.14×10~(11) CFU/mL,与感染大肠杆菌相比,斑马鱼感染副溶血弧菌后招募嗜中性粒细胞的速度更快,且数量更多。RT-qPCR结果发现斑马鱼感染副溶血弧菌后在相同时间点能够激活更高的il1b和il10基因的表达,引起强烈的炎症反应。基于以上研究,我们建立的副溶血弧菌-斑马鱼幼鱼耳泡局部感染模型,为进一步研究嗜中性粒细胞清除副溶血弧菌感染的动态过程,以及深入揭示天然免疫应答机制提供了依据。     

Transmission of Mycobacterium chelonae and Mycobacterium marinum in laboratory zebrafish through live feeds

The zebrafish (Danio rerio) is a popular vertebrate model organism used in a wide range of research fields. Mycobacteriosis, caused by Mycobacterium species, is particularly concerning because it is a common disease associated with chronic infections in these fish. Infections are also a source of uncontrolled experimental variance that may influence research results. Live feeds for zebrafish are common and include paramecia (Paramecium caudatum), brine shrimp (Artemia franciscana) and rotifers (Branchionus spp.). Although nutritionally beneficial, live feeds may pose a biosecurity risk. In this study, we investigate transmission of Mycobacterium chelonae and Mycobacterium marinum through these three live feeds. We show that all three live feeds ingest both M. marinum and M. chelonae and can transmit mycobacterial infections to zebrafish. This observation emphasizes the need for live feeds to be included in the consideration of potential biosecurity risks. This study is of importance to other beyond the zebrafish community, including those of additional aquatic models and those using live feeds for other types of aquaculture.     

二甲基联苯胺对斑马鱼早期胚胎发育的表观影响

研究了3,3'-二甲基联苯胺对斑马鱼早期胚胎发育的表观毒性影响.试验证明4.32 mg/L以上高浓度的二甲基联苯胺都能导致斑马鱼受精卵发生卵黄凝聚,使胚胎发育终止而死亡.低浓度下2.16 mg/L的二甲基联苯胺对斑马鱼受精卵的发育有一定的毒害作用,但有近一半的胚胎存活,这可能是影响其胚胎发育成活的一个临界浓度;还发现低...     

Proteomics analysis of mitochondrial extract from liver of female zebrafish undergoing starvation and refeeding

Many fish species can withstand long period of food deprivation and consequently compensate for any weight loss by undergoing rapid growth during resumption of feeding. There is an interest in taking advantage of compensatory growth to reduce feed cost. In this present study, we attempted to compare the changes in hepatic mitochondrial proteome of zebrafish undergoing starvation and refeeding. Two‐dimensional gel separation and image analysis revealed a total of 65 spots that showed changes in expression after 15 days of starvation followed by 7 days of refeeding. A total of 35 proteins were selected for mass spectrometry analysis, resulting in the positive identification of 18 proteins. Identified proteins indicated that starvation resulted in reduction in glycolysis and increase in gluconeogenesis, while refeeding caused these activities to return to normal levels. Expression pattern of several proteins related to fatty acid and amino acid metabolism also suggested the utilization of non‐carbohydrate resources for energy during starving conditions. Proteins with chaperoning and antioxidative roles such as glucose‐regulated protein, paraxonase and heat‐shock protein were also upregulated in starved conditions.     

jnk3基因在金鱼和斑马鱼不同发育时期胚胎与成体不同组织中的分化表达

JNKs是丝裂原活化蛋白激酶(MAPK)家族重要成员,参与应激反应和动物体轴的构建。为了进一步了解JNK家族在动物发育中的功能,首次分离和克隆了金鱼jnk3基因,研究了jnk3在金鱼和斑马鱼组织特异性和胚胎发育特异性表达状况。金鱼jnk3基因cDNA总长1 794 bp,其中阅读框1 293 bp,编码430个氨基酸。对jnk3基因序列及其推导的氨基酸序列进行同源性分析,结果显示,jnk3基因在脊椎动物较为保守,金鱼jnk3基因核苷酸序列与斑马鱼、人的同源性分别为94.1%和80.7%,金鱼jnk3基因氨基酸序列与斑马鱼、人的同源性分别为99.7%和93.4%。jnk3基因在鱼类成体中的表达存在着显著的组织差异,在金鱼和斑马鱼脑和精巢组织的表达具专一性,此外在斑马鱼心脏中也检测到jnk3的表达。RT-PCR检测结果显示,在鱼类胚胎发育中,神经胚期开始检测到jnk3基因mRNA表达,从神经胚到心跳期胚胎,jnk3基因表达水平逐渐增高,此后直至出膜一直保持较高的表达水平。研究表明,jnk3基因可能在鱼类后期的胚胎发育和成体脑、精巢与心脏发育中具有重要作用。     

迟缓爱德华菌人工感染斑马鱼的致病性及组织病理学变化

用迟缓爱德华菌真鲷分离株(E.t-CD)对斑马鱼进行人工感染试验,观察其发病、死亡以及相关病理学变化。结果表明,该菌对斑马鱼的半数致死浓度(LD50)为2.69×102 cfu/尾;感染发病鱼呈现岀血、溃疡、腹水、败血等症状;病理组织学检查以肝脏水肿变性,肝细胞萎缩、坏死、脱落;脾脏散在增生性结节、充血、水肿、淋巴细胞大量缺失等病变为主。结果表明,斑马鱼可作为研究迟缓爱德华菌致病性的动物模型。     

omd基因对斑马鱼骨骼矿化的影响

omd基因编码骨调蛋白,可以调控人类骨骼中钙的沉积。目前关于omd基因对鱼类骨骼钙化的作用尚不清楚,为了探究omd基因对鱼类骨骼的影响,本文调查了omd基因在斑马鱼的不同发育阶段和不同组织的表达,并利用CRISPR/Cas9基因编辑技术,成功构建斑马鱼omd基因敲除型(omd_-/-^{)。结果发现}omd基因在斑马鱼体内随着发育表达量逐渐增加,在脊柱和肌肉中表达较高；相比野生型斑马鱼,omd_-/-^{品系斑马鱼的肌肉细胞未发现明显变化,脊柱的平均钙含量下调}59.81%；与野生型相比,omd_-/-^{品系斑马鱼平均游动距离下降}33.93%,平均游动速度下降39.44%,相对静止的时间增加88.26%,游动能力下降。这些结果表明 omd基因对斑马鱼脊柱的矿化起到重要作用,并在一定程度上影响了斑马鱼的活动能力。