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Multiphoton microscopy (MPM) has become a preferred technique for intravital imaging deep in living tissues with subcellular detail, where resolution and working depths are typically optimized utilizing high numerical aperture, water-immersion objectives with long focusing distances. However, this approach requires the maintenance of water between the specimen and the objective lens, which can be challenging or impossible for many intravital preparations with complex tissues and spatial arrangements. We introduce the novel use of cohesive hyaluronan gel (HG) as an immersion medium that can be used in place of water within existing optical setups to enable multiphoton imaging with equivalent quality and far superior stability. We characterize and compare imaging performance, longevity, and feasibility of preparations in various configurations. This combination of HG with MPM is highly accessible and opens the doors to new intravital imaging applications.
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Determining the long-term cognitive impact of infections is clinically challenging. Using functional cortical connectivity, we demonstrate that interhemispheric cortical connectivity is decreased in individuals with acute Zika virus (ZIKV) encephalitis. This correlates with decreased presynaptic terminals in the somatosensory cortex. During recovery from ZIKV infection, presynaptic terminals recover, which is associated with recovered interhemispheric connectivity. This supports the contribution of synapses in the cortex to functional networks in the brain, which can be detected by widefield optical imaging. Although myeloid cell and astrocyte numbers are still increased during recovery, RNA transcription of multiple proinflammatory cytokines that increase during acute infection decreases to levels comparable to mock-infected mice during recovery. These findings also suggest that the immune response and cytokine-mediated neuroinflammation play significant roles in the integrity of brain networks during and after viral encephalitis.
Aim
We hypothesized that widefield optical imaging would allow us to assess functional cortical network disruption by ZIKV, including hippocampal-cortical networks.
Approach
We use widefield optical imaging to measure cortical functional connectivity (FC) in mice during acute infection with, and recovery from, intracranial infection with a mouse-adapted strain of ZIKV.
Results
Acute ZIKV infection leads to high levels of myeloid cell activation, with loss of neurons and presynaptic termini in the cerebral cortex and associated loss of FC primarily within the somatosensory cortex. During recovery, neuron numbers, synapses, and FC recover to levels near those of healthy mice. However, hippocampal injury and impaired spatial cognition persist. The magnitude of activated myeloid cells during acute infection predicted both recovery of synapses and the degree of FC recovery after recovery from ZIKV infection.
Conclusions
These findings suggest that a robust inflammatory response may contribute to the health of functional brain networks after recovery from infection.
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Stroke is the leading cause of chronic disability in the United States. How stroke size affects post-stroke repair and recovery is poorly understood.
Aim
We aim to investigate the effects of stroke size on early repair patterns and determine how early changes in neuronal circuits and networks predict functional outcomes after stroke.
Approach
We used wide-field optical imaging, photothrombosis, and the cylinder-rearing assay to examine changes in neuronal circuit and network activity in the context of functional recovery after stroke.
Results
Larger strokes ablating S1FP caused diffuse and widespread forepaw stimulus-evoked cortical activation, including contralesional regions evolving within 4 weeks post-stroke; smaller strokes resulted in more focused ipsilesional activation. Larger strokes decreased neuronal fidelity and bilateral coherence during stimulation of either the affected or unaffected forepaw within this 4-week period. Mice in the larger lesion group demonstrated hyperconnectivity within the contralesional hemisphere at the resting state. Greater degrees of remapping diffusivity, neuronal fidelity degradation, and hyperconnectivity predicted worse 8-week recovery after statistically controlling for the effect of infarct size.
Conclusions
These results suggest that diffuse patterns of remapping, and desynchronization and hyperconnectivity of cortical networks, evolving early after stroke may reflect maladaptive plasticity, predicting poor long-term functional recovery.
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