Project description:We report the application of low cell number sequencing of identifiable Drosophila melanogaster neurons following behavior. We demonstate the feasibility of identifying the transcriptome of 5 Mushroom Body output Neurons and 2 classes of Kenyon Cells. We find these neurons display a diverse repertoire of receptors and signaling transcripts. This information alone seems to be enough to identify each class of neurons in the study. In additional we show that aversive long-term memory induces changes in gene transcript levels in a subset of these neurons. This study provides a framework for identifying neuronal classes in Drosophila melanogaster and gaining insight into the interplay between behavior and gene regulation.
Project description:The formation of long-term memories requires changes in the transcriptional program and de novo protein synthesis. One of the critical regulators for long-term memory (LTM) formation and maintenance is the transcription factor CREB. Genetic studies have dissected the requirement of CREB activity within memory circuits, however less is known about the genetic mechanisms acting downstream of CREB and how they may contribute defining LTM phases. To better understand the downstream mechanisms, we here used a targeted DamID approach (TaDa). We generated a CREB-Dam fusion protein using the fruit fly Drosophila melanogaster as model. Expressing CREB-Dam in the mushroom bodies (MBs), a brain center implicated in olfactory memory formation, we identified genes that are differentially expressed between paired and unpaired appetitive training paradigm. Of those genes we selected candidates for an RNAi screen in which we identified genes causing increased or decreased LTM.
Project description:The Drosophila melanogaster mushroom bodies (MBs) are brain structures critical for olfactory memory. The approximately 2000 intrinsic MB neurons are divisible into alpha/beta neurons, alpha'/beta' neurons and gamma neurons by morphology and roles in memory processing. It has also been shown that the different subtypes of MB neurons have different function in the process of memory formation. This difference in function, suggested to us that different types of MB neurons might have different expression profiles. In this study, we used cell-type specific gene expression profiling to gain insight into cellular properties of MB neurons.
Project description:Mushroom bodies (MBs) are the centers for olfactory associative learning and elementary cognitive functions in the Drosophila brain. To get insights of the repertoire of MB genes that control initiation and maintenance of neural differentiation as well as the repertoire of neural factors that may have functions in the synaptic plasticity of MB neurons during learning and memory, we compared the transcript profiles between wild type and MB-ablated brains using a Drosophila whole-genome microarray. Newly hatched larvae were briefly administered with a DNA-synthesis inhibitor, hydroxyurea, and raised to adults, from which total brain RNA was analyzed. Experiment Overall Design: Two conditions analyzed: Control Brains and Musroom Body-ablated brains. Experiment Overall Design: Each condition was analyzed in triplicate.
Project description:Mushroom bodies (MBs) are the centers for olfactory associative learning and elementary cognitive functions in the Drosophila brain. To get insights of the repertoire of MB genes that control initiation and maintenance of neural differentiation as well as the repertoire of neural factors that may have functions in the synaptic plasticity of MB neurons during learning and memory, we compared the transcript profiles between wild type and MB-ablated brains using a Drosophila whole-genome microarray. Newly hatched larvae were briefly administered with a DNA-synthesis inhibitor, hydroxyurea, and raised to adults, from which total brain RNA was analyzed. Keywords: Chemical Ablation of Mushroom bodies from Drosophila brain
Project description:Obesity predisposes humans and other mammals to a range of life-threatening comorbidities, including type 2 diabetes and cardiovascular disease. Obesity and its associated dietary habits also aggravate neural pathologies, such as Alzheimer’s disease, but this class of comorbidity is less understood. When Drosophila melanogaster (flies) are exposed to high fat diet (HFD) by supplementing a standard cornmeal-sucrose-yeast medium with coconut oil, they adopt an obese phenotype of decreased lifespan, increased triglyceride storage, and hindered climbing ability. The latter development is an indicator of neurological decline in flies. Our objective was to establish the obesity-like phenotype in Drosophila and identify a correlation, if any, between obesity and neurological decline in flies through behavioral and expression microarray. We found that mated female w1118 flies exposed to HFD maintained an obese phenotype throughout adult life with onset at seven days, evidenced by increased triglyceride stores, diminished life span, and impeded climbing ability. Analysis of gene expression of the fly head via microarray and qRT-PCR validation revealed functionally relevant genes with significant fold changes. These genes had functions including in memory, metabolism, olfaction, mitosis, cell signaling, and motor function. An Aversive Phototaxis Suppression assay indicated short term memory impairment as a result of HFD. Meanwhile, there was a decline but no significant difference in odor-seeking ability with HFD. Overall, our results point to the suitability of Drosophila melanogaster to investigate connections between diet-induced obesity and nervous or neurobehavioral pathology, and to the existence of such a dynamic in an evolutionarily broad range of organisms.
Project description:This project’s aim was to compare the transcriptional profiles of olfactory sensory neurons in Drosophila melanogaster in order to identify novel genes that specify neuron-specific functions/phenotypes or may otherwise be involved in the development of the olfactory system. The isolation of sufficient numbers of intact olfactory sensory neurons (OSN) from the antenna of Drosophila melanogaster has so far limited single-cell transcriptomic approaches being applied to the adult fly antenna. Targeted DamID (TaDa) provides an alternative approach for profiling transcriptional activity in a cell-specific manor that bypasses the need for isolating OSN. Using the Gal4/UAS system, we applied TaDa to seven OSN populations and compared differences in Pol II occupancy for genes across these datasets.