Characterization of large-scale brain networks using blood-oxygenation-level-dependent functional magnetic resonance imaging is typically based on the assumption of network stationarity across the duration of scan. Recent studies in humans have questioned this assumption by showing that within-network functional connectivity fluctuates on the order of seconds to minutes. Time-varying profiles of resting-state networks (RSNs) may relate to spontaneously shifting, electrophysiological network states and are thus mechanistically of particular importance. However, because these studies acquired data from awake subjects, the fluctuating connectivity could reflect various forms of conscious brain processing such as passive mind wandering, active monitoring, memory formation, or changes in attention and arousal during image acquisition. Here, we characterize RSN dynamics of anesthetized macaques that control for these accounts, and compare them to awake human subjects. We find that functional connectivity among nodes comprising the "oculomotor (OCM) network" strongly fluctuated over time during awake as well as anaesthetized states. For time dependent analysis with short windows (<60 s), periods of positive functional correlations alternated with prominent anticorrelations that were missed when assessed with longer time windows. Similarly, the analysis identified network nodes that transiently link to the OCM network and did not emerge in average RSN analysis. Furthermore, time-dependent analysis reliably revealed transient states of large-scale synchronization that spanned all seeds. The results illustrate that resting-state functional connectivity is not static and that RSNs can exhibit nonstationary, spontaneous relationships irrespective of conscious, cognitive processing. The findings imply that mechanistically important network information can be missed when using average functional connectivity as the single network measure.
Diversity in behavioral responses to sensory stimuli has been attributed to variations in preparatory set. Variability in oculomotor responses toward identical visual stimuli has been well documented, but the neuronal processes underlying this variability are poorly understood. Here, we report evidence for set-related activity for saccadic eye movements in single neurons in the frontal eye field (FEF) in monkeys trained on a task in which they either had to look toward a visual stimulus (pro-saccade) or away from the stimulus (anti-saccade) depending on a previous instruction. A portion of FEF neurons were identified as neurons projecting directly to the superior colliculus (SC) with antidromic activation techniques. Saccade-related neurons in the FEF had lower prestimulus and stimulus-related activity on anti-saccade trials compared with pro-saccade trials. The level of prestimulus activity correlated with saccadic reaction times, express saccade occurrence, and errors in the anti-saccade task. In addition, saccade-related activity in the FEF was higher for pro-saccades than for anti-saccades. These results demonstrate that the direct descending pathway from the FEF to the SC carries preparatory set-related activity for pro-saccades and anti-saccades. The results also provide insights into the neuronal basis of variations in saccadic reaction times and in the control of the prepotent response to glance to a flashed stimulus.
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