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Physiological and morphological characterization of honeybee olfactory neurons combining electrophysiology, calcium imaging and confocal microscopy

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Abstract

The insect antennal lobe is the first brain structure to process olfactory information. Like the vertebrate olfactory bulb the antennal lobe is substructured in olfactory glomeruli. In insects, glomeruli can be morphologically identified, and have characteristic olfactory response profiles. Local neurons interconnect glomeruli, and output (projection) neurons project to higher-order brain centres. The relationship between their elaborate morphology and their physiology is not understood. We recorded electrophysiologically from antennal lobe neurons, and iontophoretically injected a calcium-sensitive dye. We then measured their spatio-temporal calcium responses to a variety of odours. Finally, we confocally reconstructed the neurons, and identified the innervated glomeruli. An increase or decrease in spiking frequency corresponded to an intracellular calcium increase or decrease in the cell. While intracellular recordings generally lasted between 10 and 30 min, calcium imaging was stable for up to 2 h, allowing a more detailed physiological analysis. The responses indicate that heterogeneous local neurons get input in the glomerulus in which they branch most strongly. In many cases, the physiological response properties of the cells corresponded to the known response profile of the innervated glomerulus. In other words, the large variety of response profiles generally found when comparing antennal lobe neurons is reduced to a more predictable response profile when the innervated glomerulus is known.

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Fig. 1A–E
Fig. 2A–G
Fig. 3A–H
Fig. 4A–D
Fig. 5A–D
Fig. 6A,B
Fig. 7A–F
Fig. 8A–E

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Abbreviations

ACT:

antenno-cerebralis-tract

AL:

antennal lobe

AP:

action potential

l-ACT:

lateral ACT

LN:

local neuron

LPL:

lateral protocerebral lobe

m-ACT:

medial ACT

MB:

mushroom body

OSN:

olfactory sensory neuron

PN:

projection neuron

T1:

tract 1 of the antennal nerve

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Acknowledgments

Thanks to Beate Eisermann for help with the figures, Astrid Klawitter for technical assistance, Robert Brandt for help with Amira, Ulrike Schröter for cell tracing, Carsten Duch for help with the electrophysiology, and Mary Wurm for help with the English. Thanks to Randolf Menzel, Silke Sachse and Ulrike Schröter for fruitful comment on the manuscript. The Volkswagenstiftung (I/75-399), the DFG (GRK 120), the BMBF (PTJ0311562X), and HFSP (RGY0050/2001) funded this work.

Animations of the calcium measurements shown in the figures can be accessed under http://galizia.ucr.edu/singleneurons

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Authors and Affiliations

  1. Institut für Biologie-Neurobiologie, Freie Universität Berlin, Königin Luise Str. 28–30, 14195, Berlin, Germany

    C. G. Galizia & B. Kimmerle

  2. Entomology 383, University of California, , Riverside, CA, 92521, USA

    C. G. Galizia

  3. Olympus BioSystems, Robert-Koch-Str 9, 82152, Planegg, Germany

    B. Kimmerle

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  1. C. G. Galizia
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Correspondence to C. G. Galizia.

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Galizia, C.G., Kimmerle, B. Physiological and morphological characterization of honeybee olfactory neurons combining electrophysiology, calcium imaging and confocal microscopy. J Comp Physiol A 190, 21–38 (2004). https://doi.org/10.1007/s00359-003-0469-0

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  • DOI: https://doi.org/10.1007/s00359-003-0469-0

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