Functional analysis of conserved C. elegans bHLH family members uncovers lifespan control by a peptidergic hub neuron.

Throughout the animal kingdom, several members of the basic helix-loop-helix (bHLH) family act as proneural genes during early steps of nervous system development. Roles of bHLH genes in specifying terminal differentiation of postmitotic neurons have been less extensively studied. We analyze here the function of 5 Caenorhabditis elegans bHLH genes, falling into 3 phylogenetically conserved subfamilies, which are continuously expressed in a very small number of postmitotic neurons in the central nervous system.

Functional analysis of conserved bHLH family members uncovers lifespan control by a peptidergic hub neuron.

Throughout the animal kingdom, several members of the basic helix-loop-helix (bHLH) family act as proneural genes during early steps of nervous system development. Roles of bHLH genes in specifying terminal differentiation of postmitotic neurons have been less extensively studied. We analyze here the function of five bHLH genes, falling into three phylogenetically conserved subfamilies, which are continuously expressed in a very small number of postmitotic neurons in the central nervous system.

Segmentation-free measurement of locomotor frequency in Caenorhabditis elegans using image invariants.

An animal's locomotor rate is an important indicator of its motility. In studies of the nematode Caenorhabditis elegans (C. elegans), assays of the frequency of body bending waves have often been used to discern the effects of mutations, drugs, or aging.

Automated multimodal imaging of behavior in multi-well plates.

Assays of behavior in model organisms play an important role in genetic screens, drug testing, and the elucidation of gene-behavior relationships. We have developed an automated, high-throughput imaging and analysis method for assaying behaviors of the nematode . We use high-resolution optical imaging to longitudinally record the behaviors of 96 animals at a time in multi-well plates, and computer vision software to quantify the animals' locomotor activity, behavioral states, and egg laying events.

Segmentation-free measurement of locomotor frequency in Caenorhabditis elegans using image invariants.

An animal's locomotor rate is an important indicator of its motility. In studies of the nematode C. elegans, assays of the frequency of body bending waves have often been used to discern the effects of mutations, drugs, or aging.

A robotic system for automated genetic manipulation and analysis of .

The nematode is one of the most widely studied organisms in biology due to its small size, rapid life cycle, and manipulable genetics. Research with depends on labor-intensive and time-consuming manual procedures, imposing a major bottleneck for many studies, especially for those involving large numbers of animals. Here, we describe a general-purpose tool, WormPicker, a robotic system capable of performing complex genetic manipulations and other tasks by imaging, phenotyping, and transferring on standard agar media.

Sleep is required to consolidate odor memory and remodel olfactory synapses.

Animals with complex nervous systems demand sleep for memory consolidation and synaptic remodeling. Here, we show that, although the Caenorhabditis elegans nervous system has a limited number of neurons, sleep is necessary for both processes. In addition, it is unclear if, in any system, sleep collaborates with experience to alter synapses between specific neurons and whether this ultimately affects behavior.

Adult-restricted gene knock-down reveals candidates that affect locomotive healthspan in C. elegans.

Understanding how we can age healthily is a challenge at the heart of biogerontological interest. Whereas myriad genes are known to affect the lifespan of model organisms, effects of such interventions on healthspan-the period of life where an animal is considered healthy, rather than merely alive-are less clear. To understand relationships between life- and healthspan, in recent years several platforms were developed with the purpose of assessing both readouts simultaneously.

Long-Term Culture and Monitoring of Isolated Caenorhabditis elegans on Solid Media in Multi-Well Devices.

The nematode Caenorhabditis elegans is among the most common model systems used in aging research owing to its simple and inexpensive culture techniques, rapid reproduction cycle (~3 days), short lifespan (~3 weeks), and numerous available tools for genetic manipulation and molecular analysis. The most common approach for conducting aging studies in C. elegans, including survival analysis, involves culturing populations of tens to hundreds of animals together on solid nematode growth media (NGM) in Petri plates.

Pharyngeal timing and particle transport defects in feeding mutants.

The nematode uses rhythmic muscle contractions (pumps) of the pharynx, a tubular feeding organ, to filter, transport, and crush food particles. A number of feeding mutants have been identified, including those with slow pharyngeal pumping rate, weak muscle contraction, defective muscle relaxation, and defective grinding of bacteria. Many aspects of these pharyngeal behavioral defects and how they affect pharyngeal function are not well understood.

An Imaging System for Monitoring C. elegans Behavior and Aging.

Many experiments in C. elegans neurobiology rely on imaging its behavior. Here we describe procedures for building a flexible and inexpensive imaging system using standard optical and mechanical components.

Optogenetic Perturbation of Individual C. elegans Pharyngeal Neurons While Monitoring Feeding Behavior.

Optogenetic approaches have proven to be powerful for examining the roles of specific neurons in generating behaviors, especially in systems where electrophysiological manipulation is not possible. Here we describe a method for optogenetically manipulating single pharyngeal neurons in intact C. elegans while monitoring pharyngeal behavior.

Cryopreservation of dehydrated with multiple recoveries using a granular medium.

Ultracold storage is widely used to preserve genetic stocks. Standard cryopreservation methods for the nematode are vulnerable to refrigeration failures, which can result in the loss of stock viability due to freeze-thaw damage. In previous work our laboratory developed a method for cryopreserving worms in a dehydrated form that remains viable after multiple freeze-thaw cycles.

Phase response analyses support a relaxation oscillator model of locomotor rhythm generation in .

Neural circuits coordinate with muscles and sensory feedback to generate motor behaviors appropriate to an animal's environment. In the mechanisms by which the motor circuit generates undulations and modulates them based on the environment are largely unclear. We quantitatively analyzed locomotion during free movement and during transient optogenetic muscle inhibition.

DAF-16/FoxO and DAF-12/VDR control cellular plasticity both cell-autonomously and via interorgan signaling.

Many cell types display the remarkable ability to alter their cellular phenotype in response to specific external or internal signals. Such phenotypic plasticity is apparent in the nematode Caenorhabditis elegans when adverse environmental conditions trigger entry into the dauer diapause stage. This entry is accompanied by structural, molecular, and functional remodeling of a number of distinct tissue types of the animal, including its nervous system.

Thermal laser ablation with tunable lesion size reveals multiple origins of seizure-like convulsions in Caenorhabditis elegans.

Laser microsurgery has long been an important means of assessing the functions of specific cells and tissues. Most laser ablation systems use short, highly focused laser pulses to create plasma-mediated lesions with dimensions on the order of the wavelength of light. While the small size of the lesion enables ablation with high spatial resolution, it also makes it difficult to ablate larger structures.

Sleep Induction by Mechanosensory Stimulation in Drosophila.

People tend to fall asleep when gently rocked or vibrated. Experimental studies have shown that rocking promotes sleep in humans and mice. However, the mechanisms underlying the phenomenon are not well understood.

Preconditioning of Caenorhabditis elegans to anoxic insult by inactivation of cholinergic, GABAergic and muscle activity.

For most metazoans, oxygen deprivation leads to cell dysfunction and if severe, death. Sublethal stress prior to a hypoxic or anoxic insult ("preconditioning") can protect cells from subsequent oxygen deprivation. The molecular mechanisms by which sublethal stress can buffer against a subsequent toxic insult and the role of the nervous system in the response are not well understood.

Dehydrated Stocks Are Resistant to Multiple Freeze-Thaw Cycles.

Ultracold preservation is widely used for storage of genetic stocks of Current cryopreservation protocols are vulnerable to refrigeration failures, which can result in the loss of stock viability due to damage during re-freezing. Here we present a method for preserving worms in a dehydrated and frozen form that retains viability after multiple freeze-thaw cycles. After dehydration in the presence of trehalose or glycerol, stocks can be frozen and thawed multiple times while maintaining viability.

A quiescent state following mild sensory arousal in Caenorhabditis elegans is potentiated by stress.

An animal's behavioral and physiological response to stressors includes changes to its responses to stimuli. How such changes occur is not well understood. Here we describe a Caenorhabditis elegans quiescent behavior, post-response quiescence (PRQ), which is modulated by the C.

Optimized criteria for locomotion-based healthspan evaluation in C. elegans using the WorMotel system.

Getting a grip on how we may age healthily is a central interest of biogerontological research. To this end, a number of academic teams developed platforms for life- and healthspan assessment in Caenorhabditis elegans. These are very appealing for medium- to high throughput screens, but a broader implementation is lacking due to many systems relying on custom scripts for data analysis that others struggle to adopt.

Quantitative imaging of sleep behavior in Caenorhabditis elegans and larval Drosophila melanogaster.

Sleep is nearly universal among animals, yet remains poorly understood. Recent work has leveraged simple model organisms, such as Caenorhabditis elegans and Drosophila melanogaster larvae, to investigate the genetic and neural bases of sleep. However, manual methods of recording sleep behavior in these systems are labor intensive and low in throughput.

A sleep state in larvae required for neural stem cell proliferation.

Sleep during development is involved in refining brain circuitry, but a role for sleep in the earliest periods of nervous system elaboration, when neurons are first being born, has not been explored. Here we identify a sleep state in larvae that coincides with a major wave of neurogenesis. Mechanisms controlling larval sleep are partially distinct from adult sleep: octopamine, the analog of mammalian norepinephrine, is the major arousal neuromodulator in larvae, but dopamine is not required.