Instrumentation for microinjection and mainteance of experimental organisms

本申请中要求的仪器将主要用于包括胚胎在内的实验生物体的显微注射，并在注射之前或之后在高度受控的条件下维持这些生物体。该设备将有助于在正常生长和发育期间或在暴露于包括疾病在内的环境和生理压力时，对生物体中的DNA、RNA和蛋白质进行表征。除了申请中提到的两名研究人员外，该设备还将满足几名研究人员的需求。麦克雷博士将分析胚胎发育和滞育过程中的基因表达，重点是蛋白质合成和功能的调节，以及分子伴侣在生理应激耐受中的作用。赖特博士将研究斑马鱼细胞内脂结合蛋白(ILBP)基因转录的机制和进化。这些研究代表了对细胞、分子和发育生物学的基本贡献，也有许多应用。例如，麦克雷博士的研究涉及心脏病发作和中风、白内障，可能还与阿尔茨海默氏症等神经系统疾病有关。对昆虫常见的生理状态--滞育的研究，可能会揭示这些生物如何抵抗压力，这在昆虫往往是害虫的农业和林业中非常重要。更好地了解滞育也将加强卤虫在水产养殖中作为饲料的使用。了解斑马鱼胚胎发育过程中的转录和基因调控将产生对理解细胞生长调控至关重要的基本信息。显然，通过使用本申请中寻求的微注射器和生长室，有可能做出对健康和商业有直接影响的发现。这些设备还将促进解决基本问题的实验，这些基本问题是许多研究计划的基石，从而使申请者及其同事在有效使用赠款资金和在几个生物学科进行现代实验的能力方面具有一定的优势。

The instrumentation requested in this application will be used primarily for microinjection of experimental organisms, including embryos, and for maintenance of these organisms under highly controlled conditions either prior to or after injection. The equipment will contribute to characterization of DNA, RNA and protein in organisms during normal growth and development or when exposed to environmental and physiological stress, including disease. The needs of several researchers in addition to the two noted in the application will be met by the equipment. Dr. MacRae will analyze gene expression during embryo development and diapause with emphasis on the regulaton of protein synthesis and function, and the contribution of molecular chaperones to the tolerance of physiological stress. Dr. Wright will study mechanistic aspects and evolution of intracellular lipid-binding protein (ILBP) gene transcription in zebrafish. These studies represent fundamental contributions to cell, molecular and developmental biology, and there are also many applications of the work. As examples, Dr. MacRae's research relates to heart attack and stroke, to cataract, and possibly to neurological disorders such as Alzheimer's. Examination of diapause, a physiological state common to insects, may reveal how these organisms resist stress, this of importance in agriculture and forestry where insects are often pests. A greater understanding of diapause will also enhance the use of Artemia as feed in aquaculture. Understanding transcription and gene regulation during zebrafish embryo development will generate fundamental information of importance in understanding regulation of cell growth. Clearly, there is potential to make discoveries which have direct consequences to health and commerce through use of the microinjector and growth chamber sought in this application. The equipment will also facilitate experiments addressing basic questions which are the cornerstone of many research programs, thus placing the applicants and their associates at a definite advantage in the effective use of grant funds and in the ability to conduct modern experimentation in several biological disciplines.