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Diversity and wiring variability of olfactory local interneurons in the Drosophila antennal lobe

Abstract

Local interneurons are essential in information processing by neural circuits. Here we present a comprehensive genetic, anatomical and electrophysiological analysis of local interneurons (LNs) in the Drosophila melanogaster antennal lobe, the first olfactory processing center in the brain. We found LNs to be diverse in their neurotransmitter profiles, connectivity and physiological properties. Analysis of >1,500 individual LNs revealed principal morphological classes characterized by coarsely stereotyped glomerular innervation patterns. Some of these morphological classes showed distinct physiological properties. However, the finer-scale connectivity of an individual LN varied considerably across brains, and there was notable physiological variability within each morphological or genetic class. Finally, LN innervation required interaction with olfactory receptor neurons during development, and some individual variability also likely reflected LN–LN interactions. Our results reveal an unexpected degree of complexity and individual variation in an invertebrate neural circuit, a result that creates challenges for solving the Drosophila connectome.

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Figure 1: Antennal lobe LNs.
Figure 2: Diversity of LN morphology.
Figure 3: Statistical analysis of glomerular innervation patterns.
Figure 4: Functional stereotypy and diversity among genetic classes.
Figure 5: Functional differences between morphological classes.
Figure 6: Variability and stereotypy of line 6 LNs.
Figure 7: Variability of patchy LNs.
Figure 8: Development but not maintenance of LN arborization depends on ORNs.

References

  1. 1

    Markram, H. et al. Interneurons of the neocortical inhibitory system. Nat. Rev. Neurosci. 5, 793–807 (2004).

    CAS  Article  Google Scholar 

  2. 2

    Olsen, S.R. & Wilson, R.I. Cracking neural circuits in a tiny brain: new approaches for understanding the neural circuitry of Drosophila. Trends Neurosci. 31, 512–520 (2008).

    CAS  Article  Google Scholar 

  3. 3

    Vosshall, L.B. & Stocker, R.F. Molecular architecture of smell and taste in Drosophila. Annu. Rev. Neurosci. 30, 505–533 (2007).

    CAS  Article  Google Scholar 

  4. 4

    Shepherd, G.M., Chen, W.R. & Greer, C.A. Olfactory Bulb. in The synaptic Organization of the Brain (ed. Shepherd, G.M.) (Oxford University Press, Oxford, 2004).

  5. 5

    Lledo, P.M., Merkle, F.T. & Alvarez-Buylla, A. Origin and function of olfactory bulb interneuron diversity. Trends Neurosci. 31, 392–400 (2008).

    CAS  Article  Google Scholar 

  6. 6

    Wachowiak, M. & Shipley, M.T. Coding and synaptic processing of sensory information in the glomerular layer of the olfactory bulb. Semin. Cell Dev. Biol. 17, 411–423 (2006).

    Article  Google Scholar 

  7. 7

    Christensen, T.A., Waldrop, B.R., Harrow, I.D. & Hildebrand, J.G. Local interneurons and information processing in the olfactory glomeruli of the moth Manduca sexta. J. Comp. Physiol. A 173, 385–399 (1993).

    CAS  Article  Google Scholar 

  8. 8

    Seki, Y. & Kanzaki, R. Comprehensive morphological identification and GABA immunocytochemistry of antennal lobe local interneurons in Bombyx mori. J. Comp. Neurol. 506, 93–107 (2008).

    CAS  Article  Google Scholar 

  9. 9

    MacLeod, K. & Laurent, G. Distinct mechanisms for synchronization and temporal patterning of odor-encoding neural assemblies. Science 274, 976–979 (1996).

    CAS  Article  Google Scholar 

  10. 10

    Fonta, C., Sun, X.J. & Masson, C. Morphology and spatial distribution of bee antennal lobe interneurons responsive to odours. Chem. Senses 18, 101–119 (1993).

    Article  Google Scholar 

  11. 11

    Ernst, K.D. & Boeckh, J. A neuroanatomical study on the organization of the central antennal pathways in insects. III. Neuroanatomical characterization of physiologically defined response types of deutocerebral neurons in Periplaneta americana. Cell Tissue Res. 229, 1–22 (1983).

    CAS  Article  Google Scholar 

  12. 12

    Stocker, R.F., Lienhard, M.C., Borst, A. & Fischbach, K.F. Neuronal architecture of the antennal lobe in Drosophila melanogaster. Cell Tissue Res. 262, 9–34 (1990).

    CAS  Article  Google Scholar 

  13. 13

    Wilson, R.I. & Laurent, G. Role of GABAergic inhibition in shaping odor-evoked spatiotemporal patterns in the Drosophila antennal lobe. J. Neurosci. 25, 9069–9079 (2005).

    CAS  Article  Google Scholar 

  14. 14

    Shang, Y., Claridge-Chang, A., Sjulson, L., Pypaert, M. & Miesenbock, G. Excitatory local circuits and their implications for olfactory processing in the fly antennal lobe. Cell 128, 601–612 (2007).

    CAS  Article  Google Scholar 

  15. 15

    Ng, M. et al. Transmission of olfactory information between three populations of neurons in the antennal lobe of the fly. Neuron 36, 463–474 (2002).

    CAS  Article  Google Scholar 

  16. 16

    Lai, S.L., Awasaki, T., Ito, K. & Lee, T. Clonal analysis of Drosophila antennal lobe neurons: diverse neuronal architectures in the lateral neuroblast lineage. Development 135, 2883–2893 (2008).

    CAS  Article  Google Scholar 

  17. 17

    Das, A. et al. Drosophila olfactory local interneurons and projection neurons derive from a common neuroblast lineage specified by the empty spiracles gene. Neural Dev. 3, 33 (2008).

    Article  Google Scholar 

  18. 18

    Okada, R., Awasaki, T. & Ito, K. Gamma-aminobutyric acid (GABA)-mediated neural connections in the Drosophila antennal lobe. J. Comp. Neurol. 514, 74–91 (2009).

    CAS  Article  Google Scholar 

  19. 19

    Lee, T. & Luo, L. Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis. Neuron 22, 451–461 (1999).

    CAS  Article  Google Scholar 

  20. 20

    Olsen, S.R., Bhandawat, V. & Wilson, R.I. Excitatory interactions between olfactory processing channels in the Drosophila antennal lobe. Neuron 54, 89–103 (2007).

    CAS  Article  Google Scholar 

  21. 21

    Lee, T., Lee, A. & Luo, L. Development of the Drosophila mushroom bodies: sequential generation of three distinct types of neurons from a neuroblast. Development 126, 4065–4076 (1999).

    CAS  Google Scholar 

  22. 22

    Jefferis, G.S.X.E., Marin, E.C., Stocker, R.F. & Luo, L. Target neuron prespecification in the olfactory map of Drosophila. Nature 414, 204–208 (2001).

    CAS  Article  Google Scholar 

  23. 23

    Jefferis, G.S. et al. Comprehensive maps of Drosophila higher olfactory centers: spatially segregated fruit and pheromone representation. Cell 128, 1187–1203 (2007).

    CAS  Article  Google Scholar 

  24. 24

    van der Goes van Naters, W. & Carlson, J.R. Receptors and neurons for fly odors in Drosophila. Curr. Biol. 17, 606–612 (2007).

    CAS  Article  Google Scholar 

  25. 25

    Hallem, E.A. & Carlson, J.R. Coding of odors by a receptor repertoire. Cell 125, 143–160 (2006).

    CAS  Article  Google Scholar 

  26. 26

    Sachse, S. et al. Activity-dependent plasticity in an olfactory circuit. Neuron 56, 838–850 (2007).

    CAS  Article  Google Scholar 

  27. 27

    Wassle, H., Peichl, L. & Boycott, B.B. Dendritic territories of cat retinal ganglion cells. Nature 292, 344–345 (1981).

    CAS  Article  Google Scholar 

  28. 28

    Grueber, W.B., Jan, L.Y. & Jan, Y.N. Tiling of the Drosophila epidermis by multidendritic sensory neurons. Development 129, 2867–2878 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. 29

    Jefferis, G.S. et al. Developmental origin of wiring specificity in the olfactory system of Drosophila. Development 131, 117–130 (2004).

    CAS  Article  Google Scholar 

  30. 30

    Sweeney, L.B. et al. Temporal target restriction of olfactory receptor neurons by Semaphorin-1a/PlexinA-mediated axon-axon interactions. Neuron 53, 185–200 (2007).

    CAS  Article  Google Scholar 

  31. 31

    Berdnik, D., Chihara, T., Couto, A. & Luo, L. Wiring stability of the adult Drosophila olfactory circuit after lesion. J. Neurosci. 26, 3367–3376 (2006).

    CAS  Article  Google Scholar 

  32. 32

    Luo, L. & Flanagan, J.G. Development of continuous and discrete neural maps. Neuron 56, 284–300 (2007).

    CAS  Article  Google Scholar 

  33. 33

    Pirez, N. & Wachowiak, M. In vivo modulation of sensory input to the olfactory bulb by tonic and activity-dependent presynaptic inhibition of receptor neurons. J. Neurosci. 28, 6360–6371 (2008).

    CAS  Article  Google Scholar 

  34. 34

    Shao, Z., Puche, A.C., Kiyokage, E., Szabo, G. & Shipley, M.T. Two GABAergic intraglomerular circuits differentially regulate tonic and phasic presynaptic inhibition of olfactory nerve terminals. J. Neurophysiol. 101, 1988–2001 (2009).

    CAS  Article  Google Scholar 

  35. 35

    Bhandawat, V., Olsen, S.R., Gouwens, N.W., Schlief, M.L. & Wilson, R.I. Sensory processing in the Drosophila antennal lobe increases reliability and separability of ensemble odor representations. Nat. Neurosci. 10, 1474–1482 (2007).

    CAS  Article  Google Scholar 

  36. 36

    Schlief, M.L. & Wilson, R.I. Olfactory processing and behavior downstream from highly selective receptor neurons. Nat. Neurosci. 10, 623–630 (2007).

    CAS  Article  Google Scholar 

  37. 37

    Miles, R. Perspectives: neurobiology. Diversity in inhibition. Science 287, 244–246 (2000).

    CAS  Article  Google Scholar 

  38. 38

    Lu, J., Tapia, J.C., White, O.L. & Lichtman, J.W. The interscutularis muscle connectome. PLoS Biol. 7, e32 (2009).

    PubMed  Google Scholar 

  39. 39

    Marin, E.C., Jefferis, G.S.X.E., Komiyama, T., Zhu, H. & Luo, L. Representation of the glomerular olfactory map in the Drosophila brain. Cell 109, 243–255 (2002).

    CAS  Article  Google Scholar 

  40. 40

    Wong, A.M., Wang, J.W. & Axel, R. Spatial representation of the glomerular map in the Drosophila protocerebrum. Cell 109, 229–241 (2002).

    CAS  Article  Google Scholar 

  41. 41

    Murthy, M., Fiete, I. & Laurent, G. Testing odor response stereotypy in the Drosophila mushroom body. Neuron 59, 1009–1023 (2008).

    CAS  Article  Google Scholar 

  42. 42

    Lichtman, J.W. & Sanes, J.R. Ome sweet ome: what can the genome tell us about the connectome? Curr. Opin. Neurobiol. 18, 346–353 (2008).

    CAS  Article  Google Scholar 

  43. 43

    Laissue, P.P. et al. Three-dimensional reconstruction of the antennal lobe in Drosophila melanogaster. J. Comp. Neurol. 405, 543–552 (1999).

    CAS  Article  Google Scholar 

  44. 44

    Hayashi, S. et al. GETDB, a database compiling expression patterns and molecular locations of a collection of Gal4 enhancer traps. Genesis 34, 58–61 (2002).

    CAS  Article  Google Scholar 

  45. 45

    Martin, J.R., Ernst, R. & Heisenberg, M. Mushroom bodies suppress locomotor activity in Drosophila melanogaster. Learn. Mem. 5, 179–191 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  46. 46

    Stocker, R.F., Heimbeck, G., Gendre, N. & de Belle, J.S. Neuroblast ablation in Drosophila P[GAL4] lines reveals origins of olfactory interneurons. J. Neurobiol. 32, 443–456 (1997).

    CAS  Article  Google Scholar 

  47. 47

    Kim, J. et al. A TRPV family ion channel required for hearing in Drosophila. Nature 424, 81–84 (2003).

    CAS  Article  Google Scholar 

  48. 48

    Dubnau, J. et al. The staufen/pumilio pathway is involved in Drosophila long-term memory. Curr. Biol. 13, 286–296 (2003).

    CAS  Article  Google Scholar 

  49. 49

    Gouwens, N.W. & Wilson, R.I. Signal propagation in Drosophila central neurons. J. Neurosci. 29, 6239–6249 (2009).

    CAS  Article  Google Scholar 

  50. 50

    Hallem, E.A., Ho, M.G. & Carlson, J.R. The molecular basis of odor coding in the Drosophila antenna. Cell 117, 965–979 (2004).

    CAS  Article  Google Scholar 

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Acknowledgements

We thank U. Heberlein (University of California, San Francisco) and E. Marin for respectively providing and screening unpublished Gal4 lines, which lead to identification of line 1 and line 6; J. Simpson (Janelia Farm, Howard Hughes Medical Institute) for providing unpublished LCCH3 (line 7) Gal4; K. Wehner (Stanford University) for mouse anti-HA; A. DiAntonio (Washington University) for rabbit anti-DVGLUT; and the Bloomington Stock Center, Kyoto Stock Center, Drosophila Genetic Resource Center, Gal4 Enhancer Trap Insertion Database (GETDB), and Developmental Studies Hybridoma Bank for other reagents. M.L.S. is grateful for the help of J. Brooks in data collection. We thank the Stanford Department of Statistics Consulting Service for technical help with statistical analyses. We thank T. Clandinin, G. Jefferis and members of the Luo and Wilson laboratories for helpful comments on the manuscript. This work was supported by US National Institutes of Health grants to L.L. (R01-DC005982) and R.I.W. (R01-DC008174), a Pew Scholar award, a McKnight Scholar award, a Sloan Foundation research fellowship, and a Beckman Young Investigator award (to R.I.W). M.L.S. is supported by a US National Research Service predoctoral award. E.Y. is supported by a Human Frontiers Science Program Long Term Fellowship. J.C.S.L. is supported by the Medical Scientist Training Program at Stanford University. L.L. receives investigator support from the Howard Hughes Medical Institute.

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Y.-H.C. and M.L.S. performed the anatomical and developmental experiments. E.Y. performed the physiological experiments. J.C.S.L. helped with statistical analysis. L.L. and R.I.W. supervised the project and wrote the paper.

Corresponding authors

Correspondence to Rachel I Wilson or Liqun Luo.

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Supplementary information

Supplementary Text and Figures

Supplementary Tables 1 and 4, Supplementary Figures 1–11 (PDF 1737 kb)

Supplementary Table 2

Raw data for ipsilateral glomerular innervation patterns (XLS 639 kb)

Supplementary Table 3

Raw data for contralateral glomerular innervation patterns of bilateral projection LNs (XLS 52 kb)

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Chou, YH., Spletter, M., Yaksi, E. et al. Diversity and wiring variability of olfactory local interneurons in the Drosophila antennal lobe. Nat Neurosci 13, 439–449 (2010). https://doi.org/10.1038/nn.2489

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