title: Multiplex assessment of the positions of odorant receptor-specific glomeruli in the mouse olfactory bulb by serial two-photon tomography.
abstract: In the mouse, axons of olfactory sensory neurons (OSNs) that express the same odorant receptor (OR) gene coalesce into one or a few glomeruli in the olfactory bulb. The positions of OR-specific glomeruli are traditionally described as stereotyped. Here, we have assessed quantitatively the positions of OR-specific glomeruli using serial two-photon tomography, an automated method for whole-organ fluorescence imaging that integrates two-photon microscopy with serial microtome sectioning. Our strategy is multiplexed. By repeated crossing, we generated two strains of mice with gene-targeted mutations at four or five OR loci for a total of six ORs: MOR23 (Olfr16), mOR37A (Olfr155), M72 (Olfr160), P2 (Olfr17), MOR256-17 (Olfr15), and MOR28 (Olfr1507). Glomerular imaging relied on intrinsic fluorescence of GFP or DsRed, or on whole-mount immunofluorescence with antibodies against GFP, DsRed, or β-gal using the method of immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO). The high-resolution 3D-reconstructed datasets were segmented to identify the labeled glomeruli and to assess glomerular positional variability between the bulbs of one mouse (intraindividual) and among the bulbs of different mice (interindividual). In 26 mice aged 21 or 50 d or 10 wk, we made measurements of the positions of 352 glomeruli. We find that positional variability of glomeruli correlates with the OR: For instance, the medial MOR28 glomerular domain occupies a surface area that is an order of magnitude larger than the surface area of the medial MOR23 glomerular domain. Our results quantify the level of precision that is delivered by the mechanisms of OSN axon wiring, differentially for the various OSN populations expressing distinct OR genes.
authors.affiliation: Max Planck Research Unit for Neurogenetics, 60438 Frankfurt, Germany; Medical Biophysics, Institute of Physiology and Pathophysiology, Faculty of Medicine, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Bolek
authors.initials: B
authors.lastname: Zapiec
authors.affiliation: Max Planck Research Unit for Neurogenetics, 60438 Frankfurt, Germany; peter.mombaerts@gen.mpg.de.
authors.forename: Peter
authors.initials: P
authors.lastname: Mombaerts
mesh_terms: D000818:Animals; D051379:Mice; D009830:Olfactory Bulb / Q000378:metabolism*; D017785:Photons; D018035:Receptors, Odorant / Q000235:genetics / Q000378:metabolism*; D014054:Tomography / Q000379:methods*
chemical_list: D018035:Receptors, Odorant
keywords: axon guidance; gene targeting; glomerulus; olfaction; olfactory sensory neuron