RESUMEN
Drosophila has become an excellent model system for investigating the organization and function of the gustatory system due to the relatively simple neuroanatomical organization of its brain and the availability of powerful genetic and transgenic technology. Thus, at the molecular and cellular levels, a great deal of insight into the peripheral detection and coding of gustatory information has already been attained. In contrast, much less is known about the central neural circuits that process this information and induce behaviorally appropriate motor output. Here, we combine functional behavioral tests with targeted transgene expression through specific driver lines to identify a single bilaterally homologous pair of bitter-sensitive interneurons that are located in the subesophageal zone of the brain. Anatomical and functional data indicate that these interneurons receive specific synaptic input from bitter-sensitive gustatory receptor neurons. Targeted transgenic activation and inactivation experiments show that these bitter-sensitive interneurons can largely suppress the proboscis extension reflex to appetitive stimuli, such as sugar and water. These functional experiments, together with calcium-imaging studies and calcium-modulated photoactivatable ratiometric integrator (CaMPARI) labeling, indicate that these first-order local interneurons play an important role in the inhibition of the proboscis extension reflex that occurs in response to bitter tastants. Taken together, our studies present a cellular identification and functional characterization of a key gustatory interneuron in the bitter-sensitive gustatory circuitry of the adult fly.
Asunto(s)
Interneuronas/fisiología , Percepción del Gusto/fisiología , Animales , Animales Modificados Genéticamente , Encéfalo/fisiología , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Interneuronas/metabolismo , Fenómenos Fisiológicos del Sistema Nervioso , Receptores de Superficie Celular/metabolismo , Células Receptoras Sensoriales/fisiología , Gusto/fisiología , Transgenes/genéticaRESUMEN
The ability to distinguish males from females is essential for productive mate selection and species propagation. Recent studies in Drosophila have identified different classes of contact chemosensory neurons that detect female or male pheromones and influence courtship decisions. Here, we examine central neural pathways in the male brain that process female and male pheromones using anatomical, calcium imaging, optogenetic, and behavioral studies. We find that sensory neurons that detect female pheromones, but not male pheromones, activate a novel class of neurons in the ventral nerve cord to cause activation of P1 neurons, male-specific command neurons that trigger courtship. In addition, sensory neurons that detect male pheromones, as well as those that detect female pheromones, activate central mAL neurons to inhibit P1. These studies demonstrate that the balance of excitatory and inhibitory drives onto central courtship-promoting neurons controls mating decisions.
Asunto(s)
Drosophila/fisiología , Vías Nerviosas/fisiología , Conducta Sexual Animal , Animales , Encéfalo/fisiología , Cortejo , Neuronas Eferentes/fisiología , Células Receptoras Sensoriales/fisiologíaRESUMEN
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca(2+) indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca(2+) detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).
Asunto(s)
Señalización del Calcio/fisiología , Calcio/metabolismo , Proteínas Luminiscentes/metabolismo , Neuronas/citología , Neuronas/fisiología , Optogenética/métodos , Animales , Rastreo Celular/métodos , Células Cultivadas , Drosophila , Luz , Proteínas Luminiscentes/genética , Microscopía Fluorescente/métodos , Ingeniería de Proteínas/métodos , Ratas , Pez CebraRESUMEN
Gustatory receptors and peripheral taste cells have been identified in flies and mammals, revealing that sensory cells are tuned to taste modality across species. How taste modalities are processed in higher brain centers to guide feeding decisions is unresolved. Here, we developed a large-scale calcium-imaging approach coupled with cell labeling to examine how different taste modalities are processed in the fly brain. These studies reveal that sweet, bitter, and water sensory cells activate different cell populations throughout the subesophageal zone, with most cells responding to a single taste modality. Pathways for sweet and bitter tastes are segregated from sensory input to motor output, and this segregation is maintained in higher brain areas, including regions implicated in learning and neuromodulation. Our work reveals independent processing of appetitive and aversive tastes, suggesting that flies and mammals use a similar coding strategy to ensure innate responses to salient compounds.
Asunto(s)
Encéfalo/citología , Encéfalo/fisiología , Neuronas/fisiología , Gusto/fisiología , Adenosina Trifosfato/farmacología , Animales , Animales Modificados Genéticamente , Encéfalo/efectos de los fármacos , Calcio/metabolismo , Relación Dosis-Respuesta a Droga , Drosophila , Proteínas de Drosophila/genética , Canales Epiteliales de Sodio , Femenino , Antagonistas del GABA/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Neuronas/efectos de los fármacos , Compuestos Organofosforados/farmacología , Receptores de Superficie Celular/metabolismo , Receptores Purinérgicos P2X2/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Sacarosa/farmacología , Edulcorantes/farmacología , Gusto/efectos de los fármacosRESUMEN
Anatomic and physiologic studies have suggested a model in which neurons of the piriform cortex receive convergent input from random collections of glomeruli. In this model, odor representations can only be afforded behavioral significance upon experience. We have devised an experimental strategy that permits us to ask whether the activation of an arbitrarily chosen subpopulation of neurons in piriform cortex can elicit different behavioral responses dependent upon learning. Activation of a small subpopulation of piriform neurons expressing channelrhodopsin at multiple loci in the piriform cortex, when paired with reward or shock, elicits either appetitive or aversive behavior. Moreover, we demonstrate that different subpopulations of piriform neurons expressing ChR2 can be discriminated and independently entrained to elicit distinct behaviors. These observations demonstrate that the piriform cortex is sufficient to elicit learned behavioral outputs in the absence of sensory input. These data imply that the piriform does not use spatial order to map odorant identity or behavioral output.