Subcellular optogenetics: Optical control of subcellular signaling and cell behavior

2^nd World Chemistry Conference

August 08-10, 2016 Toronto, Canada

Ajith Karunarathne

The University of Toledo, USA

Posters & Accepted Abstracts: Chem Sci J

Abstract :

Ability to control cell signaling is a crucial aspect of pharmacology and disease management. Lack of approaches to do so precisely in space and time in single cells to exert control over signaling and cell behavior has been an impediment. We develop and employ optogenetic approaches to control signaling in subcellular regions of single cells, visualize molecular and cellular responses and dissect signal transduction pathways of pathologically important cell behaviors such as cell migration and neuron development. Confined activation of G protein coupled receptors (GPCRs) on the plasma membrane by localized chemical gradients control cell migration. While neurite outgrowth during development is attributed to nerve growth factors (NGF) mediated activation of receptor tyrosine kinases, our findings show that G protein coupled receptors may actively involve in this process. In order to interrogate and map these pathways, we engineer light sensitive organic molecule, opsin, cryptochrome and phototropin based optogenetic-signaling triggers and employ them to optically control (i) GPCR signaling networks, (ii) specific G protein subunits and (iii) selected signaling proteins in these pathways. We employ subcellular optical control to identify signaling activities and cross talk between GPCR active leading edge and inactive trailing edge of migratory cancer cells. Our data shows that neurite outgrowth and cell migration shares G�±i/o pathway to deliver the different responses. Using our optogenetic tools, we examine how immune cells and neurons differently decode the same signal to elicit different cellular outcomes. ajith.karunarathne@utoledo.edu