Abstract
Studies of neuronal network emergence during sensory processing and motor control are greatly facilitated by technologies that allow us to simultaneously record the membrane potential dynamics of a large population of neurons in single cell resolution. To achieve whole-brain recording with the ability to detect both small synaptic potentials and action potentials, we developed a voltage-sensitive dye (VSD) imaging technique based on a double-sided microscope that can image two sides of a nervous system simultaneously. We applied this system to the segmental ganglia of the medicinal leech. Double-sided VSD imaging enabled simultaneous recording of membrane potential events from almost all of the identifiable neurons. Using data obtained from double-sided VSD imaging, we analyzed neuronal dynamics in both sensory processing and generation of behavior and constructed functional maps for identification of neurons contributing to these processes.
Highlights
One of the principal goals in neuroscience is to clarify how neuronal circuits process sensory information and control behavior
Sensory information and behavioral states are represented as dynamic activity patterns of neuronal populations in large neuronal networks
We developed a double-sided microscope for voltage-sensitive dye (VSD) imaging, consisting of precisely aligned upright and inverted fluorescent microscopes, and imaged voltage changes from the neuronal membrane stained by a highly-sensitive, fast VSD (Woodford et al, 2015)
Summary
One of the principal goals in neuroscience is to clarify how neuronal circuits process sensory information and control behavior. Sensory information and behavioral states are represented as dynamic activity patterns of neuronal populations in large neuronal networks. To clarify the neuronal mechanisms underlying sensory processing and behavioral generation, it is necessary to determine which neurons are involved in functionally relevant neuronal dynamics and how those neuronal components interact with each other within the larger network. Voltage sensors have enabled neuroscientists to examine ethologically relevant neuronal dynamics and to functionally map parts of the nervous systems of sea slugs (Bruno et al, 2015; Hill et al, 2015; Hill et al, 2014) and the medicinal leech Hirudo verbana (Briggman et al, 2005; Briggman and Kristan, 2006; Frady et al, 2016). The segmental ganglion of the leech is well suited for comprehensive recording
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