Cantrellparrish9782

The sensitivity of the internal friction to the disulfide bond status suggests that one or both of the cross-linked regions play a critical role in driving the friction-limited folding. We speculate that the molecular mechanisms giving rise to the internal friction of disulfide-reduced mSOD1 might play a role in the amyotrophic lateral sclerosis-linked aggregation of SOD1. Published by Elsevier Inc.We report the use of pulsed interleaved excitation (PIE)-fluorescence lifetime imaging microscopy (FLIM) to measure the activities of two different biosensor probes simultaneously in single living cells. Many genetically encoded biosensors rely on the measurement of Förster resonance energy transfer (FRET) to detect changes in biosensor conformation that accompany the targeted cell signaling event. One of the most robust ways of quantifying FRET is to measure changes in the fluorescence lifetime of the donor fluorophore using FLIM. The study of complex signaling networks in living cells demands the ability to track more than one of these cellular events at the same time. Here, we demonstrate how PIE-FLIM can separate and quantify the signals from different FRET-based biosensors to simultaneously measure changes in the activity of two cell signaling pathways in the same living cells in tissues. The imaging system described here uses selectable laser wavelengths and synchronized detection gating that can be tailored and optimized for each FRET pair. Proof-of-principle studies showing simultaneous measurement of cytosolic calcium and protein kinase A activity are shown, but the PIE-FLIM approach is broadly applicable to other signaling pathways. The one-dimensional information of genomic DNA is hierarchically packed inside the eukaryotic cell nucleus and organized in a three-dimensional (3D) space. Genome-wide chromosome conformation capture (Hi-C) methods have uncovered the 3D genome organization and revealed multiscale chromatin domains of compartments and topologically associating domains (TADs). Moreover, single-nucleosome live-cell imaging experiments have revealed the dynamic organization of chromatin domains caused by stochastic thermal fluctuations. However, the mechanism underlying the dynamic regulation of such hierarchical and structural chromatin units within the microscale thermal medium remains unclear. Microrheology is a way to measure dynamic viscoelastic properties coupling between thermal microenvironment and mechanical response. Here, we propose a new, to our knowledge, microrheology for Hi-C data to analyze the dynamic compliance property as a measure of rigidness and flexibility of genomic regions along with the time evolution. Our method allows the conversion of an Hi-C matrix into the spectrum of the dynamic rheological property along the genomic coordinate of a single chromosome. To demonstrate the power of the technique, we analyzed Hi-C data during the neural differentiation of mouse embryonic stem cells. We found that TAD boundaries behave as more rigid nodes than the intra-TAD regions. The spectrum clearly shows the dynamic viscoelasticity of chromatin domain formation at different timescales. Furthermore, we characterized the appearance of synchronous and liquid-like intercompartment interactions in differentiated cells. Together, our microrheology data derived from Hi-C data provide physical insights into the dynamics of the 3D genome organization. Autophagy is a key cellular degradative pathway, important for neuronal homeostasis and function. Disruption of autophagy is associated with neuronal dysfunction and neurodegeneration. Autophagy is compartmentalized in neurons, with specific stages of the pathway occurring in distinct subcellular compartments. Coordination of these stages drives progression of autophagy and enables clearance of substrates. Yet, we are only now learning how these distributed processes are integrated across the neuron. In this review, we focus on the cell biological course of autophagy in neurons, from biogenesis at the synapse to degradation in the soma. We describe how the steps of autophagy are distributed across neuronal subcellular compartments, how local machinery regulates autophagy, and the impact of coordinated regulation on neuronal physiology and disease. We also discuss how recent advances in our understanding of neuronal autophagic mechanisms have reframed how we think about the role of local regulation of autophagy in all tissues. The neural mechanisms underlying interoception-the sensation of internal physiological states-remain largely unresolved. In this issue of Neuron, Livneh et al. (2020) demonstrate that the insula bridges different timescales of interoception by tracking and predicting thirst and hunger states. By exploiting an arsenal of state-of-the-art imaging and genetic manipulation approaches, in this issue of Neuron, Marcus et al. (2020) reveal that the synapse-specific breakdown of endocannabinoid signaling in the prelimbic prefrontal cortex is a core neurobiological substrate for stress-induced, anxiety-like behaviors. In this issue of Neuron, Corkrum et al. (2020) demonstrate an unexpected role for dopamine D1 receptors on astrocytes located in the nucleus accumbens, a key structure of the brain's reward system. Activation of these receptors mediates dopamine-evoked depression of excitatory synaptic transmission, which contributes to amphetamine's psychomotor effects. Noradrenergic cells of the locus coeruleus were associated with aversive learning and arousal. In this issue of Neuron, Kaufman et al. (2020) show that they also shape the spatial map after translocation of reward. In this issue of Neuron, Sinha et al. (2020) demonstrate that synaptic organization at rod bipolar cell terminals is regulated by a leucine-rich repeat protein, LRRTM4. LRRTM4 is expressed specifically by rod bipolar cells; eliminating it in mouse retina perturbs the organization of synaptic ribbons and impairs the function of inhibitory synapses. Mosquitoes are considered to be the deadliest animals on Earth because the diseases they transmit claim at least a million human lives every year globally. Here, we discuss the scales at which the effects of ecological factors cascade to influence epidemiologically relevant behaviors of adult mosquitoes. In particular, we focused our review on the environmental conditions (coarse-scale variables) that shape the life-history traits of larvae and adult mosquitoes (fine-scale traits), and how these factors and their association, in turn, modulate adult behaviors to influence mosquito-borne disease transmission. PTC596 mw Finally, we explore the integration of physical, physiological, and behavioral information into predictive models with epidemiological applications.