Closed-loop targeted optogenetic stimulation of C. elegans populations

TL;DR

This paper introduces a high-throughput, closed-loop optogenetic system for targeted stimulation of moving C. elegans, enabling detailed behavioral analysis and significantly increasing experimental efficiency.

Contribution

The authors developed a novel system that delivers targeted, behavior-triggered optogenetic stimuli to multiple freely moving worms simultaneously, enhancing experimental throughput and precision.

Findings

Probabilistic responses depend on anterior and posterior stimulation intensities.

Stimuli during turning confirm that turning gates reversals.

Over 25-fold increase in stimulus delivery rate compared to prior methods.

Abstract

We present a high-throughput optogenetic illumination system capable of simultaneous closed-loop light delivery to specified targets in populations of moving Caenorhabditis elegans. The instrument addresses three technical challenges: it delivers targeted illumination to specified regions of the animal's body such as its head or tail; it automatically delivers stimuli triggered upon the animal's behavior; and it achieves high throughput by targeting many animals simultaneously. The instrument was used to optogenetically probe the animal's behavioral response to competing mechanosensory stimuli in the the anterior and posterior soft touch receptor neurons. Responses to more than $10^4$ stimulus events from a range of anterior-posterior intensity combinations were measured. The animal's probability of sprinting forward in response to a mechanosensory stimulus depended on both the anterior and posterior stimulation intensity, while the probability of reversing depended primarily on the posterior stimulation intensity. We also probed the animal's response to mechanosensory stimulation during the onset of turning, a relatively rare behavioral event, by delivering stimuli automatically when the animal began to turn. Using this closed-loop approach, over $10^3$ stimulus events were delivered during turning onset at a rate of 9.2 events per worm-hour, a greater than 25-fold increase in throughput compared to previous investigations. These measurements validate with greater statistical power previous findings that turning acts to gate mechanosensory evoked reversals. Compared to previous approaches, the current system offers targeted optogenetic stimulation to specific body regions or behaviors with many-fold increases in throughput to better constrain quantitative models of sensorimotor processing.