BAcTrace, a tool for retrograde tracing of neuronal circuits in Drosophila.
Journal
Nature methods
ISSN: 1548-7105
Titre abrégé: Nat Methods
Pays: United States
ID NLM: 101215604
Informations de publication
Date de publication:
12 2020
12 2020
Historique:
received:
23
01
2020
accepted:
05
10
2020
pubmed:
4
11
2020
medline:
9
2
2021
entrez:
3
11
2020
Statut:
ppublish
Résumé
Animal behavior is encoded in neuronal circuits in the brain. To elucidate the function of these circuits, it is necessary to identify, record from and manipulate networks of connected neurons. Here we present BAcTrace (Botulinum-Activated Tracer), a genetically encoded, retrograde, transsynaptic labeling system. BAcTrace is based on Clostridium botulinum neurotoxin A, Botox, which we engineered to travel retrogradely between neurons to activate an otherwise silent transcription factor. We validated BAcTrace at three neuronal connections in the Drosophila olfactory system. We show that BAcTrace-mediated labeling allows electrophysiological recording of connected neurons. Finally, in a challenging circuit with highly divergent connections, BAcTrace correctly identified 12 of 16 connections that were previously observed by electron microscopy.
Identifiants
pubmed: 33139893
doi: 10.1038/s41592-020-00989-1
pii: 10.1038/s41592-020-00989-1
pmc: PMC7610425
mid: EMS118369
doi:
Substances chimiques
Botulinum Toxins, Type A
EC 3.4.24.69
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1254-1261Subventions
Organisme : Medical Research Council
ID : MC_U105188491
Pays : United Kingdom
Organisme : NIH HHS
ID : P40 OD018537
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : European Research Council
ID : 211089
Pays : International
Organisme : NIH HHS
ID : P40 OD010949
Pays : United States
Organisme : Medical Research Council
ID : MC-U105188491
Pays : United Kingdom
Références
Luo, L., Callaway, E. M. & Svoboda, K. Genetic dissection of neural circuits: a decade of progress. Neuron 98, 256–281 (2018).
pubmed: 29673479
pmcid: 5912347
Ugur, B., Chen, K. & Bellen, H. J. Drosophila tools and assays for the study of human diseases. Dis. Model. Mech. 9, 235–244 (2016).
pubmed: 26935102
pmcid: 4833332
Venken, K. J. T., Simpson, J. H. & Bellen., H. J. Genetic manipulation of genes and cells in the nervous system of the fruit fly. Neuron 72, 202–230 (2011).
pubmed: 22017985
pmcid: 3232021
Ohyama, T. et al. A multilevel multimodal circuit enhances action selection in Drosophila. Nature 520, 633–639 (2015).
pubmed: 25896325
Zheng, Z. et al. A complete electron microscopy volume of the brain of adult Drosophila melanogaster. Cell 174, 730–743 (2018).
Takemura, S.-Y, Lu, Z. & Meinertzhagen, I. A. Synaptic circuits of the Drosophila optic lobe: the input terminals to the medulla. J. Comp. Neurol. 509, 493–513 (2008).
pubmed: 18537121
pmcid: 2481516
Talay, M. et al. Transsynaptic mapping of second-order taste neurons in flies by trans-Tango. Neuron 96, 783–795 (2017).
pubmed: 29107518
pmcid: 5693608
Huang, T.-H. et al. Tracing neuronal circuits in transgenic animals by transneuronal control of transcription (TRACT). eLife 6, e32027 (2017).
pubmed: 29231171
pmcid: 5777821
Dong, M. et al. Sv2 is the protein receptor for botulinum neurotoxin A. Science 312, 592–596 (2006).
pubmed: 16543415
Montal, M. Botulinum neurotoxin: a marvel of protein design. Annu. Rev. Biochem. 79, 591–617 (2010).
pubmed: 20233039
Pirazzini, M., Rossetto, O., Eleopra, R. & Montecucco, C. Botulinum neurotoxins: biology, pharmacology, and toxicology. Pharmacol. Rev. 69, 200–235 (2017).
pubmed: 28356439
pmcid: 5394922
Pauli, A. et al. Cell-type-specific TEV protease cleavage reveals cohesin functions in Drosophila neurons. Dev. Cell 14, 239–251 (2008).
pubmed: 18267092
pmcid: 2258333
Kubala, M. H., Kovtun, O., Alexandrov, K. & Collins, B. M. Structural and thermodynamic analysis of the GFP:GFP-nanobody complex. Protein Sci. 19, 2389–2401 (2010).
pubmed: 20945358
pmcid: 3009406
Riabinina, O. et al. Improved and expanded Q-system reagents for genetic manipulations. Nat. Methods 12, 219–222 (2015).
pubmed: 25581800
pmcid: 4344399
Washbourne, P., Pellizzari, R., Baldini, G., Wilson, M. C. & Montecucco, C. Botulinum neurotoxin types A and E require the SNARE motif in SNAP-25 for proteolysis. FEBS Lett. 418, 1–5 (1997).
pubmed: 9414082
Gupta, G. D. et al. Analysis of endocytic pathways in Drosophila cells reveals a conserved role for GBF1 in internalization via GEECs. PLoS ONE 4, e6768 (2009).
pubmed: 19707569
pmcid: 2728541
Tobin, W. F., Wilson, R. I. & Lee, W.-C. A. Wiring variations that enable and constrain neural computation in a sensory microcircuit. eLife 6, e24838 (2017).
Masse, N. Y., Turner, G. C. & Jefferis, G. S. X. E. Olfactory information processing in Drosophila. Curr. Biol. 19, R700–R713 (2009).
pubmed: 19706282
Turner, G. C., Bazhenov, M. & Laurent, G. Olfactory representations by Drosophila mushroom body neurons. J. Neurophysiol. 99, 734–746 (2008).
pubmed: 18094099
Murthy, M., Fiete, I. & Laurent, G. Testing odor response stereotypy in the Drosophila mushroom body. Neuron 59, 1009–1023 (2008).
pubmed: 18817738
pmcid: 2654402
Caron, S. J. C., Ruta, V., Abbott, L. F. & Axel., R. Random convergence of olfactory inputs in the Drosophila mushroom body. Nature 497, 113–117 (2013).
pubmed: 23615618
pmcid: 4148081
Gruntman, E. & Turner, G. C. Integration of the olfactory code across dendritic claws of single mushroom body neurons. Nat. Neurosci. 16, 1821–1829 (2013).
pubmed: 24141312
pmcid: 3908930
Nern, A., Pfeiffer, B. D., Svoboda, K. & Rubin., G. M. Multiple new site-specific recombinases for use in manipulating animal genomes. Proc. Natl Acad. Sci. USA 108, 14198–14203 (2011).
pubmed: 21831835
Aso, Y. et al. The neuronal architecture of the mushroom body provides a logic for associative learning. eLife 3, e04577 (2014).
pubmed: 25535793
pmcid: 4273437
Couto, A., Alenius, M. & Dickson., B. J. Molecular, anatomical, and functional organization of the Drosophila olfactory system. Curr. Biol. 15, 1535–1547 (2005).
pubmed: 16139208
Fishilevich, E. & Vosshall, L. B. Genetic and functional subdivision of the Drosophila antennal lobe. Curr. Biol. 15, 1548–1553 (2005).
pubmed: 16139209
Rybak, J. et al. Synaptic circuitry of identified neurons in the antennal lobe of Drosophila melanogaster. J. Comp. Neurol. 524, 1920–1956 (2016).
pubmed: 26780543
pmcid: 6680330
Klapoetke, N. C. et al. Independent optical excitation of distinct neural populations. Nat. Methods 11, 338–346 (2014).
pubmed: 24509633
pmcid: 3943671
Dolan, M.-J. et al. Communication from learned to innate olfactory processing centers is required for memory retrieval in Drosophila. Neuron 100, 651–668 (2018).
pubmed: 30244885
pmcid: 6226615
Huoviala, P. et al. Neural circuit basis of aversive odour processing in Drosophila from sensory input to descending output. Preprint at bioRxiv https://doi.org/10.1101/394403 (2018).
Dolan, M.-J. et al. Neurogenetic dissection of the Drosophila lateral horn reveals major outputs, diverse behavioural functions, and interactions with the mushroom body. eLife 8, e43079 (2019).
Butcher, N. J., Friedrich, A. B., Lu, Z., Tanimoto, H. & Meinertzhagen., I. A. Different classes of input and output neurons reveal new features in microglomeruli of the adult Drosophila mushroom body calyx. J. Comp. Neurol. 520, 2185–2201 (2012).
pubmed: 22237598
Frechter, S. et al. Functional and anatomical specificity in a higher olfactory centre. eLife 8, e44590 (2019).
Felsenberg, J. et al. Integration of parallel opposing memories underlies memory extinction. Cell 175, 709–722 (2018).
pubmed: 30245010
pmcid: 6198041
Sayin, S. et al. A neural circuit arbitrates between persistence and withdrawal in hungry Drosophila. Neuron 104, 544–558 (2019).
pubmed: 31471123
pmcid: 6839618
Kazama, H. & Wilson, R. I. Origins of correlated activity in an olfactory circuit. Nat. Neurosci. 12, 1136–1144 (2009).
pubmed: 19684589
pmcid: 2751859
Kornfeld, J. & Denk, W. Progress and remaining challenges in high-throughput volume electron microscopy. Curr. Opin. Neurobiol. 50, 261–267 (2018).
pubmed: 29758457
Schlegel, P., Costa, M. & Jefferis, G. S. X. E. Learning from connectomics on the fly. Curr. Opin. Insect Sci. 24, 96–105 (2017).
pubmed: 29208230
Bates, A. S., Janssens, J., Jefferis, G. S. X. E. & Aerts, S. Neuronal cell types in the fly: single-cell anatomy meets single-cell genomics. Curr. Opin. Neurobiol. 56, 125–134 (2019).
pubmed: 30703584
Lacy, D. B., Tepp, W., Cohen, A. C., DasGupta, B. R. & Stevens, R. C. Crystal structure of botulinum neurotoxin type A and implications for toxicity. Nat. Struct. Biol. 5, 898–902 (1998).
pubmed: 9783750
Cachero, S. & Jefferis., G. S. X. E. Drosophila olfaction: the end of stereotypy? Neuron 59, 843–845 (2008).
pubmed: 18817725
Pfeiffer, B. D. et al. Refinement of tools for targeted gene expression in Drosophila. Genetics 186, 735–755 (2010).
pubmed: 20697123
pmcid: 2942869
Pfeiffer, B. D., Truman, J. W. & Rubin, G. M. Using translational enhancers to increase transgene expression in Drosophila. Proc. Natl Acad. Sci. USA 109, 6626–6631 (2012).
pubmed: 22493255
Gibson, D. G. et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6, 343–345 (2009).
pubmed: 19363495
R Core Team. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2016).
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
pubmed: 22743772
Cachero, S. et al. BAcTrace a new tool for retrograde tracing of neuronal circuits. Zenodo https://doi.org/10.5281/zenodo.3797211 (2020).
Chiang, A.-S. et al. Three-dimensional reconstruction of brain-wide wiring networks in Drosophila at single-cell resolution. Curr. Biol. 21, 1–11 (2011).
pubmed: 21129968
Costa, M., Manton, J. D., Ostrovsky, A. D., Prohaska, S. & Jefferis, G. S. X. E. NBLAST: rapid, sensitive comparison of neuronal structure and construction of neuron family databases. Neuron 91, 293–311 (2016).
pubmed: 27373836
pmcid: 4961245
Bates, A. S. et al. Complete connectomic reconstruction of olfactory projection neurons in the fly brain. Curr. Biol. 30, 3183–3199.e6 (2020).
pubmed: 32619485
pmcid: 7443706
Kohl, J., Ostrovsky, A. D., Frechter, S. & Jefferis., G. S. X. E. A bidirectional circuit switch reroutes pheromone signals in male and female brains. Cell 155, 1610–1623 (2013).
pubmed: 24360281
pmcid: 3898676
Davletov, B., Bajohrs, M. & Binz, T. Beyond Botox: advantages and limitations of individual botulinum neurotoxins. Trends Neurosci. 28, 446–452 (2005).
pubmed: 15979165
Phan, J. et al. Structural basis for the substrate specificity of tobacco etch virus protease. J. Biol. Chem. 277, 50564–50572 (2002).
pubmed: 12377789
Jeanne, J. M., Fişek, M. & Wilson, R. I. The organization of projections from olfactory glomeruli onto higher-order neurons. Neuron 98, 1198–1213.e6 (2018).
pubmed: 29909998
pmcid: 6051339