Vivek Venkatachalam

Pan-neuronal imaging in roaming Caenorhabditis elegans.

Significance A full understanding of sensorimotor transformation during complex behaviors requires quantifying brainwide dynamics of behaving animals. Here, we characterize brainwide dynamics of individual nematodes exposed to a defined thermosensory input. We show that it is possible to uncover representations of sensory input and motor output in individual neurons of behaving animals. Panneuronal imaging in roaming animals will facilitate systems neuroscience in behaving Caenorhabditis elegans. We present an imaging system for pan-neuronal recording in crawling Caenorhabditis elegans. A spinning disk confocal microscope, modified for automated tracking of the C. elegans head ganglia, simultaneously records the activity and position of ∼80 neurons that coexpress cytoplasmic calcium indicator GCaMP6s and nuclear localized red fluorescent protein at 10 volumes per second. We developed a behavioral analysis algorithm that maps the movements of the head ganglia to the animal’s posture and locomotion. Image registration and analysis software automatically assigns an index to each nucleus and calculates the corresponding calcium signal. Neurons with highly stereotyped positions can be associated with unique indexes and subsequently identified using an atlas of the worm nervous system. To test our system, we analyzed the brainwide activity patterns of moving worms subjected to thermosensory inputs. We demonstrate that our setup is able to uncover representations of sensory input and motor output of individual neurons from brainwide dynamics. Our imaging setup and analysis pipeline should facilitate mapping circuits for sensory to motor transformation in transparent behaving animals such as C. elegans and Drosophila larva.

Local thermometry of neutral modes on the quantum Hall edge

In most electrical conductors, heat is transported by charge carriers and so both usually flow in the same direction; but in two-dimensional electron systems subject to strong magnetic fields, certain fractional quantum Hall states can cause charge and heat to flow in opposite directions.

Bidirectional thermotaxis in Caenorhabditis elegans is mediated by distinct sensorimotor strategies driven by the AFD thermosensory neurons

Significance The nematode Caenorhabditis elegans offers the opportunity to map complex behaviors to the specific roles of each neuron in a 302-neuron nervous system. Thermotaxis is a complex behavior where the worm inverts the behavioral mode—positive thermotaxis up gradients or negative thermotaxis down gradients—to move toward a remembered temperature. How are both long-term memory and multiple behavioral modes encoded? A long-standing model has been that separate circuits for positive and negative thermotaxis compete for control of body movement. In contrast, we find that different modes of thermotaxis are driven by one set of AFD thermosensory neurons. Circuits for different thermotactic behaviors diverge from the AFD neurons, probably by coupling sensory inputs to motor programs in different ways to create different thermotactic behaviors. The nematode Caenorhabditis elegans navigates toward a preferred temperature setpoint (Ts) determined by long-term temperature exposure. During thermotaxis, the worm migrates down temperature gradients at temperatures above Ts (negative thermotaxis) and performs isothermal tracking near Ts. Under some conditions, the worm migrates up temperature gradients below Ts (positive thermotaxis). Here, we analyze positive and negative thermotaxis toward Ts to study the role of specific neurons that have been proposed to be involved in thermotaxis using genetic ablation, behavioral tracking, and calcium imaging. We find differences in the strategies for positive and negative thermotaxis. Negative thermotaxis is achieved through biasing the frequency of reorientation maneuvers (turns and reversal turns) and biasing the direction of reorientation maneuvers toward colder temperatures. Positive thermotaxis, in contrast, biases only the direction of reorientation maneuvers toward warmer temperatures. We find that the AFD thermosensory neuron drives both positive and negative thermotaxis. The AIY interneuron, which is postsynaptic to AFD, may mediate the switch from negative to positive thermotaxis below Ts. We propose that multiple thermotactic behaviors, each defined by a distinct set of sensorimotor transformations, emanate from the AFD thermosensory neurons. AFD learns and stores the memory of preferred temperatures, detects temperature gradients, and drives the appropriate thermotactic behavior in each temperature regime by the flexible use of downstream circuits.

Local charge of the ν = 5/2 fractional quantum Hall state

Electrons moving in two dimensions under the influence of strong magnetic fields effectively lose their kinetic energy and display exotic behaviour dominated by Coulomb forces. When the ratio of electrons to magnetic flux quanta in the system (ν) is near 5/2, the electrons are predicted to condense into a correlated phase with fractionally charged quasiparticles and a ground-state degeneracy that grows exponentially as these quasiparticles are introduced. The only way for electrons to transform between the many ground states would be to braid the fractional excitations around each other. This property has been proposed as the basis of a fault-tolerant quantum computer. Here we present observations of localized quasiparticles at ν = 5/2, confined to puddles by disorder. Using a local electrometer to compare how quasiparticles at ν = 5/2 and ν = 7/3 charge these puddles, we were able to extract the ratio of local charges for these states. Averaged over several disorder configurations and samples, we found the ratio to be 4/3, suggesting that the local charges are = e/3 and = e/4, where e is the charge of an electron. This is in agreement with theoretical predictions for a paired state at ν = 5/2. Confirming the existence of localized e/4 quasiparticles shows that proposed interferometry experiments to test statistics and computational ability of the state at ν = 5/2 would be possible.

Local Charge of the nu=5/2 Fractional Quantum Hall State

Electrons moving in two dimensions under the influence of strong magnetic fields effectively lose their kinetic energy and display exotic behaviour dominated by Coulomb forces. When the ratio of electrons to magnetic flux quanta in the system (ν) is near 5/2, the electrons are predicted to condense into a correlated phase with fractionally charged quasiparticles and a ground-state degeneracy that grows exponentially as these quasiparticles are introduced. The only way for electrons to transform between the many ground states would be to braid the fractional excitations around each other. This property has been proposed as the basis of a fault-tolerant quantum computer. Here we present observations of localized quasiparticles at ν = 5/2, confined to puddles by disorder. Using a local electrometer to compare how quasiparticles at ν = 5/2 and ν = 7/3 charge these puddles, we were able to extract the ratio of local charges for these states. Averaged over several disorder configurations and samples, we found the ratio to be 4/3, suggesting that the local charges are = e/3 and = e/4, where e is the charge of an electron. This is in agreement with theoretical predictions for a paired state at ν = 5/2. Confirming the existence of localized e/4 quasiparticles shows that proposed interferometry experiments to test statistics and computational ability of the state at ν = 5/2 would be possible.

Contrasting responses within a single neuron class enable sex-specific attraction in Caenorhabditis elegans

Significance Roundworms carry out crucial sensory behaviors with a relatively small number of neurons. We find that male roundworms have strong preferences for particular concentrations of sex-specific small molecule cues secreted by their potential mates. These preferences emerge from the dynamics of a population of four apparently identical male-specific neurons. The response of these sensory neurons is not uniform, with some being excitatory and others inhibitory, and the timing of response varies with concentration. These features allow this single neuronal class to prefer a concentration, and potentially to calculate a derivative of chemical concentration. This previously uncharacterized neural coding strategy might allow nematodes to efficiently use a small number of cells to carry out a crucial computation to enact innate social behaviors. Animals find mates and food, and avoid predators, by navigating to regions within a favorable range of available sensory cues. How are these ranges set and recognized? Here we show that male Caenorhabditis elegans exhibit strong concentration preferences for sex-specific small molecule cues secreted by hermaphrodites, and that these preferences emerge from the collective dynamics of a single male-specific class of neurons, the cephalic sensory neurons (CEMs). Within a single worm, CEM responses are dissimilar, not determined by anatomical classification and can be excitatory or inhibitory. Response kinetics vary by concentration, suggesting a mechanism for establishing preferences. CEM responses are enhanced in the absence of synaptic transmission, and worms with only one intact CEM show nonpreferential attraction to all concentrations of ascaroside for which CEM is the primary sensor, suggesting that synaptic modulation of CEM responses is necessary for establishing preferences. A heterogeneous concentration-dependent sensory representation thus appears to allow a single neural class to set behavioral preferences and recognize ranges of sensory cues.

Gap Junctions and NCA Cation Channels Are Critical for Developmentally-Timed Sleep and Arousal in Caenorhabditis elegans

The molecular mechanisms of sleep are not fully understood. Huang et al. demonstrate that loss of Caenorhabditis elegans UNC-7 or UNC-9 innexins dramatically reduces sleep during L4/A lethargus and that those innexins are partially required... An essential characteristic of sleep is heightened arousal threshold, with decreased behavioral response to external stimuli. The molecular and cellular mechanisms underlying arousal threshold changes during sleep are not fully understood. We report that loss of UNC-7 or UNC-9 innexin function dramatically reduced sleep and decreased arousal threshold during developmentally timed sleep in Caenorhabditis elegans. UNC-7 function was required in premotor interneurons and UNC-9 function was required in motor neurons in this paradigm. Simultaneous transient overexpression of UNC-7 and UNC-9 was sufficient to induce anachronistic sleep in adult animals. Moreover, loss of UNC-7 or UNC-9 suppressed the increased sleep of EGL-4 gain-of-function animals, which have increased cyclic-GMP–dependent protein kinase activity. These results suggest C. elegans gap junctions may act downstream of previously identified sleep regulators. In other paradigms, the NCA cation channels act upstream of gap junctions. Consistent with this, diminished NCA channel activity in C. elegans robustly increased arousal thresholds during sleep bouts in L4-to-adult developmentally timed sleep. Total time in sleep bouts was only modestly increased in animals lacking NCA channel auxiliary subunit UNC-79, whereas increased channel activity dramatically decreased sleep. Loss of EGL-4 or innexin proteins suppressed UNC-79 loss-of-function sleep and arousal defects. In Drosophila, the ion channel narrow abdomen, an ortholog of the C. elegans NCA channels, drive the pigment dispersing factor (PDF) neuropeptide release, regulating circadian behavior. However, in C. elegans, we found that loss of the PDF receptor PDFR-1 did not suppress gain-of-function sleep defects, suggesting an alternative downstream pathway. This study emphasizes the conservation and importance of neuronal activity modulation during sleep, and unequivocally demonstrates that gap junction function is critical for normal sleep.