Genetic control of post-embryonic developmental progression in zebrafish

描述（由申请人提供）：R 03：斑马鱼胚胎后发育进程的遗传控制项目概述/摘要我们对脊椎动物胚胎后发育阶段的形态发生和分化机制仍然知之甚少。尽管如此，了解调控这些后期发育期的因素对于了解成人性状如何形成至关重要，并将有助于深入了解人类胚胎后胎儿和新生儿时期出现的形态缺陷和障碍。这项研究利用了斑马鱼，它经历了广泛的胚胎后发育，涉及各种器官系统的变化，这些变化与产前人类发生的过程相似或相同。这项提议采取了双管齐下的策略，以了解斑马鱼从幼虫到成虫过渡的遗传控制。第一个目标采用了有针对性的方法，集中在两个突变体，在幼虫发育过程中进行完全体细胞逮捕，逮捕后两个和三个星期的发展，分别。这些表型表明胚胎后发育进程绝对需要的基因受损;绘制和克隆突变并表征途径将揭示胚胎发生后发生的发育过程所需的一些基因。由于对胚胎后发育过程中所涉及的分子变化知之甚少，本提案的第二个目的代表了一种探索性策略，该策略将广泛表征关键发育过渡期间发生的转录变化。将为皮肤和大脑产生转录组，皮肤在胚胎后阶段经历了充分表征的组织水平转化，大脑在幼虫到成虫的转化过程中经历了广泛的神经元成熟和神经发生。此外，许多调节生长和发育的全球内分泌级联都起源于大脑。将在每个器官的不同阶段定量已知和未知mRNA（包括任何剪接变体）的转录本丰度。由于microRNA在调节无脊椎动物的阶段转换中至关重要，因此小的非编码RNA的时间表达也将被充分表征。这些胚胎后发育转录组将作为更大的研究社区的资源，并将允许检测可能介导胚胎后转变的分子和途径。动态表达谱将进一步允许发育停滞突变体的分子表型的充分表征。总体而言，这些努力将为胚胎后过渡的未来研究提供资源，并将确定调节胚胎后发育阶段正常进展所需的新因子。

DESCRIPTION (provided by applicant): R03: Genetic Control of Post-Embryonic Developmental Progression in Zebrafish Project Summary/Abstract We still very little about the mechanisms of morphogenesis and differentiation during post- embryonic developmental stages in vertebrates. Nonetheless, understanding the factors regulating these later developmental periods is essential to understanding how adult traits form, and will lend insight into morphological defects and disorders that arise during human post- embryonic fetal and neonatal periods. This research utilizes the zebrafish, which undergoes extensive post-embryonic development involving changes in a variety of organ systems, changes similar or identical to processes that occur in prenatal humans. This proposal takes a two-pronged strategy towards understanding the genetic controls of the larval-to-adult transition in zebrafish. The first aim adopts a targeted approach, concentrating on two mutants that undergo complete somatic arrest during larval development, arresting after two and three weeks of development, respectively. These phenotypes suggest an impairment of genes absolutely required for post-embryonic developmental progression; mapping and cloning the mutations and characterizing the pathways will reveal some of the genes required for developmental processes occurring after embryogenesis. Since so little is known about the molecular changes involved in post-embryonic developmental progression, the second aim of this proposal represents an exploratory strategy that will extensively characterize transcriptional changes that occur during key developmental transitions. Transcriptomes will be produced for the skin, which undergoes well-characterized tissue-level transformations at post-embryonic stages, and the brain, which undergoes extensive neuronal maturation and neurogenesis during larval-to-adult transformation. Further, many of the global endocrine cascades that regulate growth and development originate in the brain. Transcript abundance will be quantified at different stages in each organ for both known and unknown mRNAs, including any splice variants. Since microRNAs are so crucial in regulating stage transitions in invertebrates, temporal expression of small, non-coding RNAs will also be fully characterized. These post-embryonic developmental transcriptomes will serve as resources to the larger research community and will allow detection of molecules and pathways that may mediate post-embryonic transitions. Dynamic expression profiles will further allow full characterization of the molecular phenotypes of the developmental arrest mutants. Overall, these efforts will generate resources for future studies of post- embryonic transitions and will identify novel factors that regulate and are required for normal progression through post-embryonic developmental stages.