第三讲模型生物-2012.pptVIP

发育生物学Developmental Biology 大规模筛选鉴定得到的几种突变体 侧面观 头面观(腹侧) weh wt One-eyed pinhead突变体 hag wt 隐性致死突变体(weh) 显性断纹突变体(hag) （二）单倍体筛选 ♂ 依据斑马鱼的单倍体能够存活几天的特性进行诱变和快速呈现其隐性突变 (三)纯合二倍体筛选 左边－受精后最初的几分钟施以压力，抑制减数分裂第二次纺锤体形成 右边－受精后，热休克处理，抑制第一次有丝分裂，获得50％的突变体和50％野生型，胚胎的所有位点(loci)均为纯合，更便于遗传筛选 (四)病毒介导的大规模插入突变筛选 36 000 胚胎为建群鱼(P) ，选择至少3处插入的10 000多 F1代喂养，F1代彼此杂交，饲养10 000多个F2家系，每个F2家系至少进行6组姊妹交配。 从而在F3代鉴定获得大约1 000种纯合突变。 (五)行为筛选：视觉适应性突变体 斑马鱼成体night blindness杂合突变体b逃离反应的视阈比正常个体的平均值2-3 log units。 (六)荧光报告筛选 适用于活胚胎特异性酶处理过程突变筛选 Fat free 突变体幼苗胆汁荧光梯度大大减弱 小 结 斑马鱼模型系统将经典遗传学分析与胚胎观察和操作结合起来，提供了鉴定和分析调控决定动物器官发生和胚胎发育未知基因的机会。 筛选了几千个有明显异常表型的突变体，构建了胚胎发育控制基因鉴定与分析的巨大材料库。 斑马鱼创造性的遗传呈现艺术为其它模型系统的研究提供了借鉴和启迪。 斑马鱼与人类基因不仅具有很高的同源性，而且，在染色体上的基因顺序也很保守，因而，斑马鱼的遗传呈现为揭示人类发育和疾病的复杂性提供了一种行之有效的途径。 -developmental biology -genomics -development of immune system -infection of pathogens -congenitial heart disease, blood diseases -molecular neurobiology -neural development (hearing, visual, sensory systems) -ageing -apoptosis -cancer -toxicology, developmental toxicology and toxicologic pathology -drug screening As model system has been used to study A star with stripes: zebrafish as an infection model Astrid M. van der Sara, , Ben J. Appelmelka, Christina M.J.E. Vandenbroucke-Graulsa, b and Wilbert Bittera, 2004. Trends in Microbiology, 12(10): 451-457. Example 1: Development and comparison of immune system C.Elegans has a primitive innate immune system, consisting mainly of antimicrobial proteins and peptides. Drosophila immune system is more complex, but only the innate immune system seems to be present. Zebrafish have a fully developed immune system, and the adaptive immune system is an evolutionary novelty of jawed fish. This means that the interaction of a pathogen with not noly the innate but also the adaptive immune system can be studied in detail. Real-time analysis of bacterial infections in zebrafish embryos. (a) Zebrafish embryos are infected by microinjection at 28?hours postfertilization in the caudal vein (blue ar