One of the most formidable problems in neuroscience is to understand the relationship between neural activity and behavior. Behaviors are typically produced by activity in large numbers of neurons, which are often spread widely throughout the brain and spinal cord (as well as in the periphery). The patterns of neural activity during behavior have been the subject of intense study with invasive recording techniques. Systems neuroscience has experienced a revolution in the development of novel tools for imaging and recording neuronal activity in animals. These genetically encoded tools for monitoring neuronal activity capitalize upon the sequences of molecular events following neuronal activation. Each of these tools takes different approaches to capture neural activity and thus have their own unique challenges.
Fig.1 Schematic of molecular processes underlying neuronal activation. (Wang, 2019)
The very first step in action potential (AP) generation is membrane depolarization, and hence indicators of neuronal voltage provide the truest readout of neuronal activity. The best voltage indicators can resolve high-frequency APs and detect subthreshold electrical activity and hyperpolarization
Because APs reliably lead to rises in intracellular calcium, calcium imaging provides a good correlate of neural activity. Calcium indicators are evaluated by their sensitivity (large change in fluorescence attributes, or ΔF/F) and speed (desired response time is <100 ms), but other factors such as brightness, wavelength, photostability, and toxicity matter for in vivo applications as well.
pH and neurotransmitter indicators have also been developed for allowing real-time imaging of different stages of neuronal activation. The ability to sense synaptic vesicle release could enable monitoring of presynaptic activity within intact neuronal circuitry. For faithful detection of neuronal activity, these synaptic vesicle release indicators must exhibit high specificity and sensitivity, fast kinetics, good photostability, and proper targeting to synapses.
Fig.2 Schematics of genetically encoded pH and neurotransmitter indicators. (Wang, 2019)
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