|
Neuromuscular Synaptogenesis in Drosophila |
|
or centuries, a key question of interest to many has been how a synapse develops. Historically, muscles were thought to be passive structures that merely provided a substrate for the active axonal growth cone. However, we now know that faithful formation of synapses results from the precise spatiotemporal interactions of the molecular components responsible for target guidance and recognition, cytoskeletal remodeling and subsynaptic specialization. This project investigates the in vivo cellular dynamics that are suspected to have major impacts on the process of synaptogenesis. Using the Drosophila neuromuscular junction as a model, my research attempts to gain a better understanding of the three-dimensional interactions in vivo between synaptic partners, which are currently unknown. Available high-resolution bio-imaging techniques (GFP- and RFP-based bio-probes together with time-lapse confocal microscopy) allows me to perform in vivo analysis at high cellular resolutions using live, undissected organisms. This intact embryo context preserves the spatial three-dimensionality of those molecular and physical factors playing a role in axon guidance, target recognition and synaptogenesis. It also facilitates the study of mechanisms involved in brain organization within their native environment. GFP (green fluorescent protein) and RFP (red fluorescent protein) tagged proteins are being visualized using different acquisition techniques. Depending on the purpose of the experiment, 2D time-lapse (high temporal resolution), 3D (high spatial resolution) or 4D (3D time-lapse; a hybrid level resolution) image stacks are acquired using inverted microscopes with either laser scanning or spinning-disc confocal modules. The image stacks are then analyzed using various 3D reconstruction and deconvolution software packages. In order to facilitate the organization of this site and to facilitate discussion of this highly specific and complicated developmental event, I have divided this part of my research in three main areas. Nevertheless, the three remain linked and inter-dependent (see figure below) in many ways. Myopodia (post-synaptic filopodia) Neuromuscular synaptogenesis (Muscle Innervation) In summary, this project investigates the in vivo cellular dynamics that are suspected to have major impacts of the process of synaptogenesis. The results are anticipated to enhance our hitherto underappreciated cellular dynamics that surround, or perhaps even dictate the process of synaptogenesis. |
|
F |
|
Contact Info: |
|
Developmental timeline with approximate timing of events from the axon, muscle and muscle myopodia perspective. |
|
by Franklin Carrero-Martínez |
|
© This site is protected by copyright laws under US and International law. All rights reserved. © 2004 Franklin Carrero-Martínez, Dr. Akira Chiba |