Visualization of axonal protein allocation in Drosophila with whole-brain localization microscopy

Long-term memory (LTM) formation requires learning-induced protein synthesis in specific neurons and synapses within a neural circuit. Precisely how neural activity allocates new proteins to specific synaptic ensembles, however, remains unknown. We developed a deep-tissue super-resolution imaging tool suitable for single-molecule localization in intact adult Drosophila brain, and focused on the axonal protein allocation in mushroom body (MB), a central neuronal structure involved in olfactory memory formation. We found that insufficient training suppresses LTM formation by inducing the synthesis of vesicular monoamine transporter (VMAT) proteins within a dorsal paired medial (DPM) neuron, which innervates all axonal lobes of the MB. Surprisingly, using our localization microscopy, we found that these learning-induced proteins are distributed only in a subset of DPM axons in specific sectors along the MB lobes. This neural architecture suggests that sector-specific modulation of neural activity from MB neurons gates consolidation of early transient memory into LTM.

[1]  Paola Cognigni,et al.  Do the right thing: neural network mechanisms of memory formation, expression and update in Drosophila , 2018, Current Opinion in Neurobiology.

[2]  Maximilian T. Strauss,et al.  Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT , 2017, Nature Communications.

[3]  A. Chiang,et al.  Long-term memory requires sequential protein synthesis in three subsets of mushroom body output neurons in Drosophila , 2017, Scientific Reports.

[4]  Louis K. Scheffer,et al.  A connectome of a learning and memory center in the adult Drosophila brain , 2017, eLife.

[5]  Robert H Singer,et al.  Quantitative mRNA imaging throughout the entire Drosophila brain , 2016, Nature Methods.

[6]  I. Katona,et al.  Presynaptic Protein Synthesis Is Required for Long-Term Plasticity of GABA Release , 2016, Neuron.

[7]  Yoshinori Aso,et al.  Dopaminergic neurons write and update memories with cell-type-specific rules , 2016, eLife.

[8]  S. Sprecher,et al.  Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue , 2016, Front. Cell. Neurosci..

[9]  Wesley R. Legant,et al.  High density three-dimensional localization microscopy across large volumes , 2016, Nature Methods.

[10]  Min Zhang,et al.  Dissecting neural pathways for forgetting in Drosophila olfactory aversive memory , 2015, Proceedings of the National Academy of Sciences.

[11]  Gerald M. Rubin,et al.  Plasticity-driven individualization of olfactory coding in mushroom body output neurons , 2015, Nature.

[12]  Dheeraj S. Roy,et al.  Memory engram storage and retrieval , 2015, Current Opinion in Neurobiology.

[13]  Jeff W. Lichtman,et al.  Clarifying Tissue Clearing , 2015, Cell.

[14]  Edward S. Boyden,et al.  Expansion microscopy , 2015, Science.

[15]  Ronald L. Davis,et al.  Aging Impairs Protein-Synthesis-Dependent Long-Term Memory in Drosophila , 2015, The Journal of Neuroscience.

[16]  G. Rubin,et al.  The neuronal architecture of the mushroom body provides a logic for associative learning , 2014, eLife.

[17]  Guy M. Hagen,et al.  ThunderSTORM: a comprehensive ImageJ plug-in for PALM and STORM data analysis and super-resolution imaging , 2014, Bioinform..

[18]  Liang Gao,et al.  3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy , 2014, Nature Protocols.

[19]  E. Kandel,et al.  The Molecular and Systems Biology of Memory , 2014, Cell.

[20]  S. Waddell.  Reinforcement signalling in Drosophila; dopamine does it all after all , 2013, Current Opinion in Neurobiology.

[21]  Sjoerd Stallinga,et al.  Measuring image resolution in optical nanoscopy , 2013, Nature Methods.

[22]  Ann-Shyn Chiang,et al.  Drosophila ORB protein in two mushroom body output neurons is necessary for long-term memory formation , 2013, Proceedings of the National Academy of Sciences.

[23]  Ann-Shyn Chiang,et al.  Systems memory consolidation in Drosophila , 2013, Current Opinion in Neurobiology.

[24]  Alexander Y Katsov,et al.  Fast and sensitive multi-color 3D imaging using aberration-corrected multi-focus microscopy , 2012, Nature Methods.

[25]  Chenglong Xia,et al.  Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes , 2012, Proceedings of the National Academy of Sciences.

[26]  Yoshinori Aso,et al.  Three Dopamine Pathways Induce Aversive Odor Memories with Different Stability , 2012, PLoS genetics.

[27]  A. Chiang,et al.  Visualizing Long-Term Memory Formation in Two Neurons of the Drosophila Brain , 2012, Science.

[28]  A. Rohrbach,et al.  Propagation stability of self-reconstructing Bessel beams enables contrast-enhanced imaging in thick media , 2012, Nature Communications.

[29]  A. Diaspro,et al.  Live-cell 3D super-resolution imaging in thick biological samples , 2011, Nature Methods.

[30]  Bing Zhang,et al.  Elevated Levels of the Vesicular Monoamine Transporter and a Novel Repetitive Behavior in the Drosophila Model of Fragile X Syndrome , 2011, PloS one.

[31]  Atsushi Miyawaki,et al.  Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain , 2011, Nature Neuroscience.

[32]  Tzumin Lee,et al.  Serotonin–mushroom body circuit modulating the formation of anesthesia-resistant memory in Drosophila , 2011, Proceedings of the National Academy of Sciences.

[33]  Jianyong Tang,et al.  Three-Dimensional Super-resolution Imaging of Thick Biological Samples , 2009, Microscopy and Microanalysis.

[34]  Tim Tully,et al.  Excess protein synthesis in Drosophila Fragile X mutants impairs long-term memory , 2008, Nature Neuroscience.

[35]  M. Heilemann,et al.  Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes. , 2008, Angewandte Chemie.

[36]  E. Betzig,et al.  Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics , 2008, Nature Methods.

[37]  Mark Bates,et al.  Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy , 2008, Science.

[38]  Yueqing Peng,et al.  Dopamine-Mushroom Body Circuit Regulates Saliency-Based Decision-Making in Drosophila , 2007, Science.

[39]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[40]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[41]  Michael J. Krashes,et al.  Drosophila Dorsal Paired Medial Neurons Provide a General Mechanism for Memory Consolidation , 2006, Current Biology.

[42]  Ronald L. Davis,et al.  Drosophila DPM Neurons Form a Delayed and Branch-Specific Memory Trace after Olfactory Classical Conditioning , 2005, Cell.

[43]  R. Greenspan,et al.  Dopaminergic Modulation of Arousal in Drosophila , 2005, Current Biology.

[44]  Thomas Preat,et al.  Exclusive Consolidated Memory Phases in Drosophila , 2004, Science.

[45]  Jay Hirsh,et al.  Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase. , 2003, Journal of neurobiology.

[46]  A. Chiang,et al.  Three‐dimensional mapping of brain neuropils in the cockroach, Diploptera punctata , 2001, The Journal of comparative neurology.

[47]  S. Hell,et al.  Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. , 1994, Optics letters.

[48]  K. Tsurugi,et al.  RNA N-glycosidase activity of ricin A-chain. Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. , 1987, The Journal of biological chemistry.

[49]  K. Tsurugi,et al.  The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28 S ribosomal RNA caused by the toxins. , 1987, The Journal of biological chemistry.