Live Imaging of the Genesis and Differentiation of the Sympathetic Nervous System

交感神经系统起源和分化的实时成像

• 批准号: 0719251
• 负责人: Frances Lefcort
• 金额: $ 39.09万
• 依托单位: Montana State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0719251&HistoricalAwards=false
• 关键词: Live; Imaging; Genesis; Differentiation; Sympathetic

A major focus in biology is to understand the cellular and molecular mechanisms that orchestrate the morphogenesis of a developing embryo. Traditionally, most of this analysis has been conducted on fixed tissue, which although informative, does not allow for the capture of the dynamic cellular movements that mediate morphogenetic events. Given these constraints, it has become increasingly recognized in the field that live imaging can provide us with an extremely powerful mechanism for elucidating multiple key biological events. A fascinating developing system to which this technology can be applied, is the migrating neural crest. The neural crest is a heterogeneous population of progenitor cells that migrate throughout the vertebrate embryo and give rise to a plethora of key derivatives, including the vast majority of the neurons and glia of the peripheral nervous system, the cartilage and mesectododerm of the face, and the melanocytes of the skin. This system has been a major topic of study for over 100 years and these studies have contributed significantly to our understanding of stem cells, the effects of environmental cues on cellular differentiation and the role of prepatterning, intrinsic cues in specifiying cell fate. Because of the critical role the autonomic nervous system plays in maintaining homeostasis of all vertebrates, it is this system which we will investigate in this proposal. With the advent of fluorescent probes, migrating cells can be followed in living tissue for hours or days. From these data, scientists have developed new views of how neural crest cells migrate and form their distinctive derivatives. In addition, in the chick embryo there are numerous mechanisms for conducting gain-and-loss of function molecular perturbations. Thus, the approach is to combine these two powerful technologies in order to elucidate the cellular and molecular mechanisms that mediate the formation of the sympathetic nervous system from their precursor neural crest cells. The experiments proposed here provide an excellent educational training opportunity for both the undergraduate and graduate students who will conduct the proposed experiments, and data gained from these studies will be discussed and analyzed in the undergraduate courses the PI teaches on campus. Furthermore the PI often brings elementary students to the lab so that they can experience the thrill of watching live cells move in a real live embryo on the confocal microscope. There is no more effective means to make science immediate and tangible for the non-scientist than first hand experience in the lab.

生物学的一个主要重点是了解细胞和分子机制，协调发育胚胎的形态发生。 传统上，大多数这种分析已经进行了固定的组织，虽然信息，不允许捕获的动态细胞运动介导的形态发生事件。 鉴于这些限制，它已成为越来越多地认识到，在现场成像可以为我们提供一个非常强大的机制，阐明多个关键的生物事件。 这项技术可以应用于一个迷人的发展系统，是迁移神经嵴。 神经嵴是一种异质的祖细胞群体，其在整个脊椎动物胚胎中迁移并产生过多的关键衍生物，包括外周神经系统的绝大多数神经元和神经胶质、面部的软骨和中外胚层以及皮肤的黑素细胞。 这个系统已经是一个主要的研究课题超过100年，这些研究已经大大有助于我们的干细胞的理解，环境线索对细胞分化的影响和预图案化的作用，内在的线索在specifying细胞命运。由于自主神经系统在维持所有脊椎动物的内环境稳定方面起着关键作用，因此我们将在本建议中研究该系统。 随着荧光探针的出现，迁移的细胞可以在活组织中跟踪数小时或数天。从这些数据中，科学家们对神经嵴细胞如何迁移并形成其独特的衍生物提出了新的观点。 此外，在鸡胚中，有许多机制进行功能分子扰动的获得和丧失。因此，该方法是联合收割机这两个强大的技术，以阐明细胞和分子机制，介导的交感神经系统的形成，从他们的前体神经嵴细胞。这里提出的实验提供了一个很好的教育培训的机会，为本科生和研究生谁将进行拟议的实验，从这些研究中获得的数据将在PI在校园里教的本科课程进行讨论和分析。 此外，PI经常带小学生到实验室，让他们体验在共聚焦显微镜上观察真实的活胚胎中活细胞运动的刺激。对于非科学家来说，没有比在实验室中获得第一手经验更有效的方法来使科学变得直接和有形。