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Freifeld, L.

Publications and source records attributed to Freifeld, L..

2 recordsLinked to original sources

Establishing an assay for visual desensitization in larval zebrafish for understanding synaptic plasticity

Synaptic plasticity, the ability of neurons to modify the strength of their connections, is essential for learning. Desensitization is a type of synaptic plasticity during which a behavioral response to a stimulus is reduced after repeated presentations of a weakened version of the stimulus which does not produce the behavioral response and thus is deemed a subthreshold stimulus. Here we explore whether the larval zebrafish escape response from visual threats, such as an expanding dark spot, can act as a potential model to study desensitization. We developed a novel experimental setup to investigate visual desensitization, exposing larvae to looming visual threats of varying contrast, and used high-speed cameras to verify rapid escape movements. First, we established stimuli which reliably acted as subthreshold and suprathreshold stimuli. Next, we quantified the response rate before and after desensitization and compared those data to control larvae who were not desensitized. We observed desensitized larvae reduced their response rate to threatening looming stimuli. By providing an opportunity to explore the plasticity mechanisms governing sensory-motor transformations and behavioral adaptation, this research contributes to our understanding of fundamental principles of learning and neural plasticity across vertebrates.

neuroscience↗

Precision calcium imaging of dense neural populations via a cell body-targeted calcium indicator

Methods for one-photon fluorescent imaging of calcium dynamics in vivo are popular due to their ability to simultaneously capture the dynamics of hundreds of neurons across large fields of view, at a low equipment complexity and cost. In contrast to two-photon methods, however, one-photon methods suffer from higher levels of crosstalk between cell bodies and the surrounding neuropil, resulting in decreased signal-to-noise and artifactual correlations of neural activity. Here, we address this problem by engineering cell body-targeted variants of the fluorescent calcium indicator GCaMP6f. We screened fusions of GCaMP6f to both natural as well as engineered peptides, and identified fusions that localized GCaMP6f to within approximately 50 microns of the cell body of neurons in live mice and larval zebrafish. One-photon imaging of soma-targeted GCaMP6f in dense neural circuits reported fewer artifactual spikes from neuropil, increased signal-to-noise ratio, and decreased artifactual correlation across neurons. Thus, soma-targeting of fluorescent calcium indicators increases neuronal signal fidelity and may facilitate even greater usage of simple, powerful, one-photon methods of population imaging of neural calcium dynamics.

neuroscience↗