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

Montana State University, Bozeman MT

Investigators

Abstract

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.