Name: measuring and altering mating drive in male drosophila melanogaster
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Description: Watch this Scientific Journal Video about Measuring and Altering Mating Drive in Male Drosophila melanogaster at JoVE.com

The authors thank Mike Crickmore, Dragana Rogulja, and Michelle Frank for comments on the manuscript. Pavel Gorelik provided technical support for manufacturing the behavioral arenas. This work was conducted in Mike Crickmore's lab and is also supported by the Whitehall Foundation (Principal Investigator: Dragana Rogulja). S.X.Z. is a Stuart H.Q. and Victoria Quan Fellow at Harvard Medical School.

NOTA: Este protocolo describe la preparación (secciones 1 - 3), ejecución (Sección 4), y el análisis (Sección 4) de los ensayos de saciedad en 2-D. Luego, utilizando la estimulación dopaminérgica como ejemplo, la sección 5 muestra la forma de combinar la estimulación thermogenetic con los ensayos de saciedad 2-D para inducir hipersexualidad. Sección 6 describe 3 maneras de verificar los resultados de los ensayos de saciedad en 2-D. Por último, la sección 7 muestra cómo medir la recuperación de la unidad de...

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Estados motivacionales pueden ser saciados, mantienen, y se recuperaron 34. Presentamos un ensayo de saciedad 2-D de forma rápida y robusta que mide todos estos aspectos de apareamiento unidad en la marcha. Este ensayo se abre la posibilidad de utilizar manipulaciones genéticas mosca avanzada para estudiar los componentes moleculares y de circuito de un comportamiento motivado. El ensayo de saciedad se basa en la capacidad del varón a la corte con éxito y copular,...

El trabajo en Drosophila ha proporcionado la penetración profunda y pionera en muchos fenómenos biológicos, incluyendo la naturaleza del gen 1, los principios del desarrollo embrionario 2, 3 ritmos circadianos, y el desarrollo y el cableado del sistema nervioso 4, 5, 6. La motivación sigue siendo mucho menos conocido que estos fenómenos, tal vez debido a las limitaciones de los sistemas que se han estudiado hasta el momento. La motivación en la mosca se estudia principalmente en el contexto de hambre, que presenta muchos retos debido a su extremadamente pequeña ingesta de alimentos por combate alimentación y exoesqueleto que impide signos manifiestos de la deposición de grasa. En consecuencia, hay una necesidad de ampliar los sistemas utilizados para estudiar la motivación en la marcha. Se describe un marco de comportamiento para el estudio de la unidad de acoplamiento enDrosophila. Este sistema se aprovecha de las herramientas neurogenéticas en la mosca, así como la accesibilidad 7, 8, 9, 10, 11, 12 y la conectoma putativo de su circuito de dimorfismo sexual 8, 13. Además, gran parte de la innata 14, 15, 16, 17, 18, 19, 20, 21 y aprendimos 22, 23, 24 circuitos sensorio-motor que controla el cortejo se ha trabajado de manera detallada, proporcionando una oportunidad únicapara localizar el nodo de circuito exacta sobre la que incide la motivación. Recientemente hemos informado de que, en la marcha, al igual que en los seres humanos, los niveles de dopamina son fundamentales para la unidad de acoplamiento 25, 26, 27. Hemos tenido acceso a la genética a nivel de circuito molecular- y detallada productora de dopamina y la recepción de las neuronas en la marcha, lo que facilita los análisis pertinentes de este fenómeno conservada usando los ensayos que aquí describimos 25. Añadimos a los ensayos de comportamiento en Zhang et al. 25 una nueva arena de comportamiento plana que permite la creación de vídeo, lo que llamamos un ensayo de saciedad de 2 dimensiones (2-D), una mejora importante sobre los métodos anteriores. En consecuencia, el nuevo ensayo es más escalable y cuantificable, y por tanto más adecuado para pantallas genéticas de los genes y las neuronas implicadas en la motivación. Utilizamos este nuevo ensayo, junto con ensayos de cortejo y neurogemanipulaciones néticas, para demostrar cómo medir y alterar la unidad de acoplamiento de la marcha.

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measuring and altering mating drive in male drosophila melanogaster

A pesar de décadas de investigación, las bases neuronales y moleculares de los estados motivacionales siguen siendo un misterio. Recientemente hemos desarrollado una novela, reduccionista, y un sistema escalable para la investigación en profundidad de la motivación de utilizar la unidad de acoplamiento macho de Drosophila melanogaster (Drosophila), los métodos para los que detallamos aquí. El paradigma de comportamiento se centra en el descubrimiento de que la unidad de apareamiento masculino disminuye junto con la fertilidad a lo largo de cópulas repetidas y recupera más de ~ 3 d. En este sistema, las herramientas neurogenéticas potentes disponibles en la mosca convergen con la accesibilidad genética y diagrama de cableado putativo disponible para el comportamiento sexual. Esta convergencia permite un rápido aislamiento e interrogatorio de pequeñas poblaciones neuronales con funciones específicas de motivación. A continuación detallamos el diseño y ejecución del ensayo de saciedad que se utiliza para medir la motivación y alterar el cortejo en la mosca macho. Usando estoensayo, también demostramos que la unidad de apareamiento masculino baja puede ser superada mediante la estimulación de las neuronas dopaminérgicas. El ensayo de saciedad es simple, asequible y robusta a las influencias de los antecedentes genéticos. Esperamos que el ensayo de saciedad para generar muchos nuevos conocimientos sobre la neurobiología de los estados motivacionales.

Para la caracterización de la unidad de apareamiento de Drosophila, 3 días de edad, WT Cantón-S machos se probaron en un ensayo de saciedad 2-D. En el transcurso del ensayo (4,5 h), los machos se aparean un promedio de 4,8 ± 0,3 (media ± error estándar de la media, SEM) veces. Apareamientos inician sobre todo en las primeras 2 h (78%) (Figura 6A, 6B) y se vuelven menos frecuentes a medida que avanza el ensayo (Figura 6A, 6B). Esta disminuc...

Watch this Scientific Journal Video about Measuring and Altering Mating Drive in Male Drosophila melanogaster at JoVE.com

Measuring and Altering Mating Drive in Male Drosophila melanogaster

En este artículo se describe un ensayo de comportamiento que utiliza la unidad de apareamiento masculino en Drosophila melanogaste R a estudiar la motivación. Usando este método, los investigadores pueden utilizar técnicas de neurogenéticas mosca avanzada para descubrir los mecanismos genéticos, moleculares y celulares que subyacen a esta motivación.

La medición y la modificación de acoplamiento de unidad en Male<em&gt; Drosophila melanogaster</em

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, ...

Research

JoVE Journal

Neuroscience

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Grabaciones solo Sensillum en los insectos Drosophila melanogaster Y Anopheles gambiae</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

Investigación

Neurociencias

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenético estimulación de comportamiento de escape en<em&gt; Drosophila melanogaster</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determinación de la actividad locomotora espontánea en<em&gt; Drosophila melanogaster</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Ensayo sencillo para cuantificación de Prevención Social en<em&gt; Drosophila melanogaster</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Teñido Insectos de la Conducta ensayos: el comportamiento de apareamiento de anestesiada<em&gt; Drosophila</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

Conditions Affecting Social Space in Drosophila melanogaster

Conditions Affecting Social Space in Drosophila melanogaster

Condiciones que afectan Espacio Social en<em&gt; Drosophila melanogaster</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Forrajeo Protocolo Ruta de longitud de<em&gt; Drosophila melanogaster</em&gt; Las larvas

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

El alimentador de ensayo capilar para analizar el consumo de alimentos en<em&gt; Drosophila melanogaster</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Análisis metabólico del cerebro adulto y Larval de Drosophila melanogaster

This protocol describes a method for measuring the metabolism in Drosophila melanogaster larval and adult brains. Quantifying metabolism in whole organs ...

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Evaluación morfológica y funcional de los axónyis y sus sinapsis durante la muerte de Axon en Drosophila melanogaster

Axon degeneration is a shared feature in neurodegenerative disease and when nervous systems are challenged by mechanical or chemical forces. However, our ...