斑马鱼资源的开发保藏与国家斑马鱼资源中心

斑马鱼是一种新兴的脊椎模式动物。在过去的30年中,斑马鱼已被广泛应用于生命科学、健康科学、环境农业等诸多科研领域。为了满足不同的科研需要,研究人员开发和利用各种技术创建了大量的斑马鱼基因突变和转基因品系,这些品系已成为开展相关科学研究的宝贵资源。为了更好地保藏和利用这些资源,在全球范围内建设有多个规模不一的斑马鱼资源库。2012年,我国的国家斑马鱼资源中心（http：//zfish.cn）在中国科学院水生生物研究所正式成立。本文将重点介绍全球斑马鱼资源的开发和保藏情况,以及我国国家斑马鱼资源中心的最新建设进展。     

模型动物斑马鱼及其特定病原净化

虽然斑马鱼等实验用鱼已经成为生命科学研究的一类重要模型动物,但对于在集约化人工养殖过程中实验鱼的疾病却研究很少。实验用斑马鱼严重的健康问题使其在研究中常出现大面积甚至毁灭性的死亡。常见的感染也可以危及很多实验室的鱼群。因此,培育健康的实验用鱼群,建立必要的质量控制标准已经势在必行。本文收集和分析了大量相关的资料,为我国实验用鱼疾病的控制以及标准化研究和培育SPF斑马鱼提供帮助。     

实验用鱼遗传质量控制及标准化

在遗传学及其他生命科学研究领域, 实验用鱼已成为一类应用越来越广的实验动物, 但是尚缺少标准化的质量控制标准和监管。在我国, 实验动物实行严格的许可证制度和质量监督制度。实验用鱼遗传质量控制标准是实验用鱼质量控制的基础。为了规范实验用鱼的遗传质量, 避免实验用鱼种质退化、遗传漂移, 导致实验结果误差, 开展了本标准的研究。依据《实验动物管理条例》, 参考国内外实验用鱼遗传学相关的研究成果, 结合我国实验用鱼生产和使用的实际情况, 在全面收集、分析实验数据和广泛征询专家意见的基础上, 以实验用斑马鱼和剑尾鱼遗传质量控制为规范对象, 研究制定了实验用鱼遗传质量控制标准, 供科研工作者参考、讨论。本标准对实验用斑马鱼和剑尾鱼的遗传分类及命名原则、实验用鱼的繁殖方法、近交系和封闭群的遗传质量监测进行了规范。新标准将为实验用鱼的使用和管理提供理论支撑。     

斑马鱼基因工程的研究进展

对国内外斑马鱼基因工程研究进展。包括最近获得的大批转基因斑马鱼品系、靶向筛选的转基因斑马鱼、斑马鱼转基因技术的发展和斑马鱼基因工程的热点问题与应用前景进行了综述。指出我国已建立日趋成熟的斑马鱼转基因技术、细胞核移植技术和染色体组操作技术,具有斑马鱼细胞遗传学和胚胎学基础,我国有望在不久的将来获得定向整合的基因工程斑马鱼,并推动生物技术应用研究领域的进步和发展。     

生长激素促进斑马鱼尾鳍的再生

目的建立生长激素过表达的转基因斑马鱼,研究生长激素在斑马鱼尾鳍再生过程中的作用。方法利用Gateway技术构建表达质粒"pDestTol2CG2; ubi:GH-polyA",在一细胞期显微注射表达质粒和转座酶mRNA后,通过荧光显微镜和qPCR技术筛选鉴定GH过表达的转基因斑马鱼。将斑马鱼分为对照组(野生型)和生长激素过表达组,尾鳍横切后,记录分析斑马鱼尾鳍再生过程。结果转基因斑马鱼中心脏被绿色荧光蛋白标记。荧光定量PCR检测结果显示GH表达水平显著高于对照组(P<0.05)。斑马鱼尾鳍横断后,生长激素过表达组的再生速度显著提高(P<0.05)。结论建立稳定生长激素过表达的转基因斑马鱼品系,过表达生长激素能够提高斑马鱼尾鳍再生速度。     

斑马鱼脊柱区的发育

斑马鱼(Danio rerio)作为模式生物,有关它的研究不胜枚举,然而对其脊柱区发育的相关研究却相对较少。本文从中枢神经系统发育、骨骼发育、骨骼肌发育、相关结缔组织及脊柱区的运动5个方面对斑马鱼脊柱区的发育进行综述,以便进一步认识这一优良动物模型,并对其相关基础研究起到启示作用。     

台湾地区斑马鱼研究现况

台湾地区斑马鱼研究起始于1996年,在经历了约8年的萌芽期(1996~2003年)之后,目前已进入到茁壮期(2004~现今),现今全台湾共有83个实验室使用斑马鱼作为实验材料,台湾地区斑马鱼研究社群的研究主题可大致分成3大类:(1)胚胎发育;(2)人类疾病;(3)生物技术。累积至今与斑马鱼相关论文发表总数已达到342篇。自2010年起,台湾也成立了两个斑马鱼种质资源库(TZCAS与ZeTH)。在种质资源库的充分协助下,目前许多医院的临床医师、工程与生物信息相关领域的研究人员,也开始加入斑马鱼研究社群进行跨领域的整合性研究,成为现今台湾地区斑马鱼研究的一大特色。     

MicroRNAs对斑马鱼发育的调控

microRNAs(miRNAs)是一类长度约为22nt的非编码小RNA,从单细胞到多细胞真核生物中都广泛存在,在进化过程中高度保守,对动物发育、生理功能及病理过程都具有重要调控作用。斑马鱼(Danio rerio)是现代生物学研究中广泛使用的模式动物,以斑马鱼为模型研究miRNAs可以揭示miRNAs在脊椎动物中的功能。文章就miRNAs整体缺失对斑马鱼胚胎发育的影响及一些miRNAs在斑马鱼早期发育过程中的调控机制进行了综述,从而为探索miRNAs在脊椎动物中的功能及鱼类的生产育种提供理论基础。     

斑马鱼:在生命科学中畅游

模式动物是生命科学研究的重要材料,为生命科学的发展做出了重要的贡献。自20世纪90年代初以来,斑马鱼因其多方面的优点已成为模式动物家族中重要的一员,受到越来越多的重视和利用。目前,斑马鱼被广泛地用于发育生物学、遗传学、肿瘤学、药物学、毒理与环保等方面的研究,不断涌现新的研究成果。     

Manganese and cobalt activate zebrafish ovarian cancer G-protein-coupled receptor 1 but not GPR4

Mammalian ovarian G-protein-coupled receptor 1 (OGR1) is activated by some metals in addition to extracellular protons and coupling to multiple intracellular signaling pathways. In the present study, we examined whether zebrafish OGR1, zebrafish GPR4, and human GPR4 (zOGR1, zGPR4, and hGPR4, respectively) could sense the metals and activate the intracellular signaling pathways. On one hand, we found that only manganese and cobalt of the tested metals stimulated SRE-promoter activities in zOGR1-overexpressed HEK293T cells. On the other hand, none of the metals tested stimulated the promoter activities in zGPR4- and hGPR4-overexpressed cells. The OGR1 mutant (H4F), which is lost to activation by extracellular protons, did not stimulate metal-induced SRE-promoter activities. These results suggest that zOGR1, but not GPR4, is also a metal-sensing G-protein-coupled receptor in addition to a proton-sensing G-protein-coupled receptor, although not all metals that activate hOGR1 activated zOGR1.     

脑膜炎球菌多糖疫苗培养基的标准化研究

目的用干粉原料完全替代以消化液为主要原料的培养基配方,实现脑膜炎球菌多糖疫苗培养基的标准化生产。方法用筛选改良的酪蛋白酸水解物干粉替代50%盐酸酪蛋白水解液,以改良酵母浸出粉替代酵母透析液和酵母浸出粉,适当调整培养基配方,改进并稳定培养基制备工艺,确定适宜质量指标,并对各项质量指标数据进行分析。结果改进后的脑膜炎球菌多糖疫苗干粉培养基有效地降低了批间差异,各项质量指标均符合脑膜炎球菌培养要求,培养基的各项质量指标更加稳定可控,在生产中得到了良好、稳定的生长结果。结论用干粉原料完全替代脑膜炎球菌多糖疫苗培养基中的消化液和酵母透析液是成功的,同时改良的干粉培养基进一步明确了配方标准,使原料准备、制备工艺、质量指标控制更加标准化,有效提高了培养基质量,有利于规模化生产,促进了脑膜炎球菌多糖疫苗生产用培养基的标准化工作。     

斑马鱼超弱光特性的初步研究

本文报道有关鱼类(斑马鱼)超弱发光的新资料:光谱分布、发光动力学曲线及有关参数与pH,O_2,剥鳞和化学物质对超弱发光的影响,以及鱼体表面积和鱼类检测时的精度和准度。     

Sodium selenite/selenium nanoparticles (SeNPs) protect cardiomyoblasts and zebrafish embryos against ethanol induced oxidative stress

Alcoholic cardiomyopathy is the damage caused to the heart muscles due to high level of alcohol consumption resulting in enlargement and inflammation of the heart. Selenium is an important trace element that is beneficial to human health. Selenium protects the cells by preventing the formation of free radicals in the body. In the present study, protein mediated synthesis of SeNPs was investigated. Two different sizes of SeNPs were synthesized using BSA and keratin. The synthesized SeNPs were characterized by scanning electron microscopy (SEM) with elemental composition analysis Energy Dispersive X-ray spectroscopy(EDX) and X-ray diffraction (XRD). This study demonstrates the in vitro and in vivo antioxidative effects of sodium selenite and SeNPs. Further selenium and SeNPs were evaluated for their ability to protect against 1% ethanol induced oxidative stress in H9C2 cell line. The selenium and SeNPs were found to reduce the 1% ethanol-induced oxidative damage through scavenging intracellular reactive oxygen species. The selenium and SeNPs could also prevent pericardial edema induced ethanol treatment and reduced apoptosis and cell death in zebrafish embryos. The results indicate that selenium and SeNPs could potentially be used as an additive in alcoholic beverage industry to control the cardiomyopathy.     

Inhibitory control in zebrafish,Danio rerio

We assessed whether zebrafish, Danio rerio, display inhibitory control using a simple and rapid behavioural test. Zebrafish were exposed to a prey stimulus placed inside a transparent tube, which initially elicited attack behaviour. However, zebrafish showed a rapid reduction in the number of attacks towards the prey, which indicated the ability to inhibit their foraging behaviour. Zebrafish also exhibited mnemonic retention of foraging inhibition, as indicated by a reduced number of attacks in a subsequent exposure to the unreachable prey. The ability to inhibit the foraging behaviour varied across three genetically separated wild-type strains and across different individuals within strains, suggesting that zebrafish show heritable within-species differences in inhibitory control. Our behavioural test might be suitable for screening large zebrafish populations in mutational studies and assessing the effects of pharmacologically active substances on inhibitory control.     

Autotaxin regulates vascular development via multiple lysophosphatidic acid (LPA) receptors in zebrafish

Autotaxin (ATX) is a multifunctional ecto-type phosphodiesterase that converts lysophospholipids, such as lysophosphatidylcholine, to lysophosphatidic acid (LPA) by its lysophospholipase D activity. LPA is a lipid mediator with diverse biological functions, most of which are mediated by G protein-coupled receptors specific to LPA (LPA1-6). Recent studies on ATX knock-out mice revealed that ATX has an essential role in embryonic blood vessel formation. However, the underlying molecular mechanisms remain to be solved. A data base search revealed that ATX and LPA receptors are conserved in wide range of vertebrates from fishes to mammals. Here we analyzed zebrafish ATX (zATX) and LPA receptors both biochemically and functionally. zATX, like mammalian ATX, showed lysophospholipase D activity to produce LPA. In addition, all zebrafish LPA receptors except for LPA5a and LPA5b were found to respond to LPA. Knockdown of zATX in zebrafish embryos by injecting morpholino antisense oligonucleotides (MOs) specific to zATX caused abnormal blood vessel formation, which has not been observed in other morphant embryos or mutants with vascular defects reported previously. In ATX morphant embryos, the segmental arteries sprouted normally from the dorsal aorta but stalled in midcourse, resulting in aberrant vascular connection around the horizontal myoseptum. Similar vascular defects were not observed in embryos in which each single LPA receptor was attenuated by using MOs. Interestingly, similar vascular defects were observed when both LPA1 and LPA4 functions were attenuated by using MOs and/or a selective LPA receptor antagonist, Ki16425. These results demonstrate that the ATX-LPA-LPAR axis is a critical regulator of embryonic vascular development that is conserved in vertebrates.     

黍稷的名实考证及规范

黍稷是起源于中国最古老的作物,糯者为黍,粳者为稷。长期以来对黍稷的称谓一直不能统一,特别是对稷的称谓在农史界一直争论不休,难以定论。本文从黍稷的起源演化过程、细胞学和遗传学的研究、黍稷是人类最早驯化的作物、我国甲骨文和古农书的记载等方面,论证了黍稷的起源早于粟。并以此为据,进一步辩证了稷不是粟,同时指出将黍稷称为软糜和硬糜、糜黍、糜子、黍子等不同称谓的不足之处,进而认为以黍稷作为规范称谓为好。     

Keratodont formulae in anuran tadpoles: proposals for a standardization

The various systems proposed to describe concisely, in the form of a formula, the number and arrangement of keratodont rows in the oral disk of anuran tadpoles, are reviewed and classified. A distinction is made between individual and collective keratodont formulae. The different systems are compared and evaluated, and proposals are made to standardize the use of various signs, numerals and letters in keratodont formulae. Nine standardized systems (seven all-cardinal and two cardinal-ordinal systems) are finally retained: these should cover all needs according to the kind of use and the level of precision required.     

Culture of cells from zebrafish (Brachydanio rerio) blastula-stage embryos

The zebrafish has become a popular model for studies of vertebrate development and toxicology. However,in vitro approaches utilizing this organism have not been fully exploited due to the absence of suitable cell culture systems. Previously, we developed methods for the culture of cells derived from zebrafish blastula-stage embryos. One of these cultures, ZEM-2, was derived in a complex medium containing trout embryo extract, trout serum and medium conditioned by buffalo rat liver cells. In this study we describe a zebrafish embryo cell line, ZEM-2A, derived from ZEM-2 following selection for growth in a simplified medium. Optimal growth of ZEM-2A cells is attained in nutrient medium supplemented with 5% fetal bovine serum.