Effects of Anxiety on Anticipation and Visual Search in Dynamic, Time-Constrained Situations

We tested the predictions of Attentional Control Theory (ACT) by examining how anxiety affects visual search strategies, performance efficiency, and performance effectiveness using a dynamic, temporal-constrained anticipation task. Higher and lower skilled players viewed soccer situations under 2 task constraints (near vs. far situation) and were tested under high (HA) and low (LA) anxiety conditions. Response accuracy (effectiveness) and response time, perceived mental effort, and eye-movements (all efficiency) were recorded. A significant increase in anxiety was evidenced by higher state anxiety ratings on the MRF-L scale. Increased anxiety led to decreased performance efficiency because response times and mental effort increased for both skill groups whereas response accuracy did not differ. Anxiety influenced search strategies, with higher skilled players showing a decrease in number of fixation locations for far situations under HA compared with LA condition when compared with lower skilled players. Findings provide support for ACT with anxiety impairing processing efficiency and, potentially, top-down attentional control across different task constraints.

A versatile toolkit to produce sensitive FRET biosensors to visualize signaling in time and space

Genetically encoded, ratiometric biosensors based on fluorescence resonance energy transfer (FRET) are powerful tools to study the spatiotemporal dynamics of cell signaling. However, many biosensors lack sensitivity. We present a biosensor library that contains circularly permutated mutants for both the donor and acceptor fluorophores, which alter the orientation of the dipoles and thus better accommodate structural constraints imposed by different signaling molecules while maintaining FRET efficiency. Our strategy improved the brightness and dynamic range of preexisting RhoA and extracellular signal-regulated protein kinase (ERK) biosensors. Using the improved RhoA biosensor, we found micrometer-sized zones of RhoA activity at the tip of F-actin bundles in growth cone filopodia during neurite extension, whereas RhoA was globally activated throughout collapsing growth cones. RhoA was also activated in filopodia and protruding membranes at the leading edge of motile fibroblasts. Using the improved ERK biosensor, we simultaneously measured ERK activation dynamics in multiple cells using low-magnification microscopy and performed in vivo FRET imaging in zebrafish. Thus, we provide a construction toolkit consisting of a vector set, which enables facile generation of sensitive biosensors.