Fluorescent stereomicroscope, microinjection training module and objective upgrade for C. elegans transgenic generation, husbandry and phenotypic screening

器官是细胞的功能单位，在生物体内执行特定的任务。胚胎发育研究中的一个重要问题是不同类型的细胞如何迁移、改变形状并以协调的方式相互作用形成一个器官。在我的实验室里，我们使用秀丽线虫，一种自由生活的蛔虫，以及它强大的遗传学和活体成像工具来研究器官发育。特别是，我们研究了蠕虫前肠的唾液腺细胞如何利用与前肠其他细胞的相互作用来形成它们的细长形状。我们的研究将结合多个个体细胞在空间和时间上的相互作用，形成器官发育的4维模型。与细胞培养研究相比，这种方法将提供器官发育的视角，更接近地模拟其真正的复杂性。早期器官发育中微小的形态发生变化会影响最终的器官结构，因此可以为器官功能和结构的进化提供一种机制。线虫遗传学的强大工具将确定推动这一变化的分子途径。这项研究的长期目标是将这些基本发现应用于植物寄生线虫的高度专门化前景的开发，即与农业相关的马铃薯胞囊线虫。对我的研究项目来说，至关重要的是产生转基因动物的能力。在转基因线虫的产生过程中，唾液腺发育所需基因的DNA被重新引入动物体内。基因序列的修饰被用来探测该基因的功能。秀丽线虫作为一种模式生物，也提供了基因筛查的能力，可以识别调节唾液腺发育的重要但未确定特征的基因的突变。基因筛选和转基因菌株饲养需要高倍率的荧光立体显微镜。确定亚细胞结构的变化需要具有高倍率物镜的显微镜。转基因动物的产生需要一个显微注射系统。所要求的设备对我的研究计划的推进和对学生进行先进显微镜和显微技术的培训是必不可少的。

Organs are functional units of cells that perform specific tasks within an organism. An important question in the study of embryological development is how distinct cell types migrate, change shape and interact in a coordinated manner to form an organ. In my laboratory we use Caenorhabditis elegans, a free-living roundworm, with its powerful tools of genetics and live-imaging to study organ development. In particular, we study how the salivary gland cells of the worm's foregut use interactions with other cells of the foregut to form their elongated shape. Our research will combine multiple individual cellular interactions in space and time to form a 4-dimensional model of organ development. When compared to cell culture studies, this approach will provide a perspective of organ development that more closely models its true complexity. Small morphogenic changes in early organ development can impact final organ structure and, therefore, can provide a mechanism for the evolution of organ function and structure. The powerful tool of C. elegans genetics will identify the molecular pathways that are the driving force behind this change. The long-term goal of the research is to apply these basic findings to the development of the highly specialized foregut of the plant-parasitic nematodes, i.e., the agriculturally relevant Potato Cyst Nematode. Essential to my research program is the ability to generate transgenic animals. During the generation of transgenic nematodes, the DNA of genes required for salivary gland development is reintroduced into the animal. Modification of a gene's sequence is used to probe the function of that gene. C. elegans as a model organism also offers the power of genetic screens to identify mutations in important but uncharacterized genes regulating salivary gland development. Genetic screens and transgenic strain husbandry require a high-magnification fluorescent stereomicroscope. Defining changes in subcelluar structure require microscopes with high magnification objectives. The generation of transgenic animals requires a microinjection system. The requested equipment is essential for the advancement of my research program and the training of students in advanced microscopy and microtechniques.