nematode worm culture in dog food agar plates for behavioral analysis

Investigación de comportamientos colectivos en nematodos utilizando medio de agar para alimentos para perros

Both nonscientists and scientists are fascinated with animals&#39; collective behaviors, as in flocks of birds and schools of fish. Collective behaviors have been analyzed in a broad range of fields, including physics, biology, mathematics, and robotics. In particular, active matter physics is a growing research field that focuses on systems composed of self-propelled elements, that is, dissipative systems, such as flocks of birds, schools of fish, biofilms of motile bacteria, cytoskeletons composed of active molecules, and groups of self-propelled colloids. The theory of active matter physics maintains that however complex the behaviors of individuals are, the collective motions of enormous numbers of living things are governed by a small number of simple rules. For example, the Vicsek model, a candidate for a unified description of the collective motion of self-propelled particles, predicts that short-range alignment interaction of moving objects is required to form a long-range ordered phase with eccentric fluctuation in 2D, as in herds of animals1. Top-down experimental approaches pertaining to the physics of active matter are developing rapidly. Previous experiments confirmed the formation of a long-range ordered phase in Escherichia coli2. Other recent works&#160;employed cells3,4, bacteria5, motile colloids6, or moving proteins7,8. Simple minimal models such as the Vicsek model successfully described these real phenomena. In contrast with these unicellular experimental systems, collective behaviors by animals are usually observed in the wild, as no one could hope to perform controlled experiments with 10,000 real birds or fish.
Biologists share the same interest as physicists: how individuals interact with each other and functionally behave as a group. One of the traditional research fields for analyzing individual behavior is neuroscience, in which the mechanisms underlying behavior have been examined at the neuronal and molecular levels. Many neuroscientific bottom-up approaches have been developed thus far. Top-down approaches in physics and bottom-up approaches in biology can be facilitated using model animals such as the fruit fly, the worm Caenorhabditis elegans, and the mouse9. However, there have been few findings on the large-scale collective behavior of these model animals in the laboratory10; it is still difficult to prepare genetically tractable model animals on a large scale in the laboratory. Therefore, in current research on collective behaviors in biology and physics, it has been difficult for scientists who usually do research in the laboratory to study animals&#39; collective behaviors.
In this study, we established a method for the large-scale cultivation of nematodes to study their collective behaviors. This system allows us to change environmental conditions and examine the effect of individual-level locomotion on collective behaviors using mutants10. In active matter physics, the parameters of the mathematical model can be controlled in both experiments and simulations, which enables verification of that model for identifying unified descriptions. Genetics is used to understand the neural circuit mechanism underlying collective behavior11.

Autor destacado: Análisis colectivo del comportamiento del nematodo, Caenorhabditis elegans

El cultivo a gran escala de nematodos para estudiar sus comportamientos colectivos

We are studying the mechanism underlying collective behavior from the perspective of active metaphysics and neuroscience. In the research field of collective behavior, experimental system has been established using non-living things and at the cellular level. However, it remains still challenging to design the animal-level experimental system to study the collective behavior in the laboratory.We successfully reconstructed the collective behavior with traditionally used model animal, nematode C.elegans. This system make it possible to control the parameter with mathematical model proposed for animals'collective behavior, and studies the neuromechanism underlying collective behavior using molecular genetics.

Cultivo de gusanos nematodos en placas de agar para alimentos para perros para análisis de comportamiento

Comience colocando cuatro gusanos adultos en un medio de crecimiento de nematodos o en una placa de agar NGM sembrada con E. coli OP50. Cultive los gusanos de primera generación hasta la inanición en la placa NGM a una temperatura de 23 grados centígrados durante siete días. A continuación, transfiera pequeñas cantidades de agar para comida para perros o medio DFA al centro de una nueva placa NGM sembrada con E. coli OP50.Para experimentos optogenéticos, vierta 40 microlitros de todo trans-retinal en el DFA antes de la inoculación del gusano. A continuación, 0,5 miligramos de comida para perros en el medio DFA, a unos dos milímetros de distancia de la tapa del plato. Evite la contaminación irradiando la placa NGM con luz ultravioleta durante 15 minutos.Reúna los gusanos hambrientos de las placas NGM con agua esterilizada. A continuación, inocule los gusanos recolectados en el medio DFA ubicado en el NGM. Propague los gusanos a 23 grados centígrados, lo que les permitirá trepar hacia la tapa de la placa durante 10 a 14 días.El aumento de la humedad exhibió tamaños de compartimentos más grandes de los patrones de la red de lombrices antes de colapsar finalmente, dejando grupos de lombrices latentes en la superficie interna de la tapa.

Research

JoVE Journal

Biology

Nematode Worm Culture in Dog Food Agar Plates for Behavioral Analysis

The Large-Scale Cultivation of Nematodes to Study Their Collective Behaviors

Animals exhibit dynamic collective behaviors, as observed in flocks of birds, schools of fish, and crowds of humans. The collective behaviors of animals ...

Investigación

Biología

Observation of the Collective Behavior and Optogenetic Experimentation of Nematode Dauer Worms

Observación del comportamiento colectivo y experimentación optogenética de gusanos nematodos Dauer