Structure-Function Mapping of Olfactory Bulb Circuits with Synchrotron X-ray Nanotomography
Information is routed between brain areas via parallel streams. Neurons may share common inputs yet convey distinct information to different downstream targets. Here, we leverage the anatomical organisation of the mouse olfactory bulb (OB), where dozens of projection neurons (mitral and tufted cells, M/TCs) affiliate with a single input unit, a glomerulus. To link functional properties of M/TCs to their anatomical glomerular association at scale, we combine in vivo two-photon (2P) imaging with synchrotron CT anatomical analysis and targeted X-ray nano-holotomography (XNH). Improving XNH resolution for mm3 volumes enables us to reliably identify subcellular features, automatically segment >80,000 cell nuclei in individual experiments, and delineate several hundred functionally imaged projection neurons and their detailed morphology, including up to 20 M/TCs per individual glomerulus ('sister' cells). In over 2400 sister cell pairs, we consistently find that odour response amplitudes to a panel of 47 monomolecular odours are conserved between sister cells, with, however, distinct responses to individual odours. Responses correlated with anatomical features such as cell body position and lateral dendritic arborisation. Thus, sister cells neither simply relay glomerular inputs nor are they dominated by network activity. Instead, they show a 'balanced diversity' in their responses, enabling efficient encoding of odour stimuli whilst retaining the overall structure of odour space. Thus, synchrotron X-ray tomography can reliably link subcellular anatomy to function in a non-destructive way across the mm3 scale. With recent advances in X-ray optics and the emergence of 4th generation synchrotrons, it becomes conceivable to extend this highly accessible approach to entire brain regions with increasing resolution.