Acquisition of confocal scan to upgrade existing fluorescence microscope

University Of Kentucky, Lexington KY

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Abstract

Project Summary This proposal is for an equipment administrative supplement to an existing MIRA R35 supporting the PI’s research addressing the molecular mechanisms of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) signal transmission as guided by scaffold proteins. This research program focuses on understanding the mechanisms by which key regulators, such as the scaffold protein Shoc2, modify ERK1/2 signaling. These studies decipher and determine the mechanisms by which endosomal sorting regulates the ubiquitin-driven remodeling of Shoc2 complexes and signaling and illuminate the key steps that segregate Shoc2 into distinct intracellular sorting pathways. Preliminary studies have already yielded new insights into the molecular details of how Shoc2-activated ERK1/2 signals promote early developmental stages. For this work, zebrafish is used as a vertebrate model to investigate the in vivo effect of Shoc2 gene editing on the early stages of development since they offer distinct advantages for studies of development, including their transparency, as well as external and rapid development. Outcomes from this work are expected to produce an ambitious and comprehensive mechanistic understanding of how Shoc2 is involved in determining the specificity of ERK1/2 signaling outcomes. As such, this work is laying the groundwork for future studies on developmental disorders that will contribute to the advancement of novel therapeutic strategies and biomarkers. This administrative supplement application to acquire a confocal scanning disk as an ‘add-on’ to an existing 3i Marianas microscope platform will foster ongoing experiments in this research program. The advanced scanning capacities will significantly improve the quality of the imaging analysis to examine the distribution of Shoc2 signaling complexes in cells and will dramatically enhance these studies to examine the role of Shoc2- mediated signals in embryonic development.