Glomerular Map Research Articles

Dimensionality reduction simplifies synaptic partner matching in an olfactory circuit.

The distribution of postsynaptic partners in three-dimensional (3D) space presents complex choices for a navigating axon. Here, we discovered a dimensionality reduction principle in establishing the 3D glomerular map in the fly antennal lobe. Olfactory receptor neuron (ORN) axons first contact partner projection neuron (PN) dendrites at the 2D spherical surface of the antennal lobe during development, regardless of whether the adult glomeruli are at the surface or interior of the antennal lobe. Along the antennal lobe surface, axons of each ORN type take a specific 1D arc-shaped trajectory that precisely intersects with their partner PN dendrites. Altering axon trajectories compromises synaptic partner matching. Thus, a 3D search problem is reduced to 1D, which simplifies synaptic partner matching and may generalize to the wiring process of more complex brains.

Roles of odorant receptors during olfactory glomerular map formation.

The organization of the olfactory glomerular map involves the convergence of olfactory sensory neurons (OSNs) expressing the same odorant receptor (OR) into glomeruli in the olfactory bulb (OB). A remarkable feature of the olfactory glomerular map formation is that the identity of OR instructs the topography of the bulb, resulting in thousands of discrete glomeruli in mice. Several lines of evidence indicate that ORs control the expression levels of various kinds of transmembrane proteins to form glomeruli at appropriate regions of the OB. In this review, we will discuss how the OR identity is decoded by OSNs into gene expression through intracellular regulatory mechanisms.

Shaping the olfactory map: cell type-specific activity patterns guide circuit formation.

The brain constructs spatially organized sensory maps to represent sensory information. The formation of sensory maps has traditionally been thought to depend on synchronous neuronal activity. However, recent evidence from the olfactory system suggests that cell type-specific temporal patterns of spontaneous activity play an instructive role in shaping the olfactory glomerular map. These findings challenge traditional views and highlight the importance of investigating the spatiotemporal dynamics of neural activity to understand the development of complex neural circuits. This review discusses the implications of new findings in the olfactory system and outlines future research directions.

Insertion of a neomycin selection cassette in the Amigo1 locus alters gene expression in the olfactory epithelium leading to region-specific defects in olfactory receptor neuron development.

During development of the nervous system, neurons connect to one another in a precisely organized manner. Sensory systems provide a good example of this organization, whereby the composition of the outside world is represented in the brain by neuronal maps. Establishing correct patterns of neural circuitry is crucial, as inaccurate map formation can lead to severe disruptions in sensory processing. In rodents, olfactory stimuli modulate a wide variety of behaviors essential for survival. The formation of the olfactory glomerular map is dependent on molecular cues that guide olfactory receptor neuron axons to broad regions of the olfactory bulb and on cell adhesion molecules that promote axonal sorting into specific synaptic units in this structure. Here, we demonstrate that the cell adhesion molecule Amigo1 is expressed in a subpopulation of olfactory receptor neurons, and we investigate its role in the precise targeting of olfactory receptor neuron axons to the olfactory bulb using a genetic loss-of-function approach in mice. While ablation of Amigo1 did not lead to alterations in olfactory sensory neuron axonal targeting, our experiments revealed that the presence of a neomycin resistance selection cassette in the Amigo1 locus can lead to off-target effects that are not due to loss of Amigo1 expression, including unexpected altered gene expression in olfactory receptor neurons and reduced glomerular size in the ventral region of the olfactory bulb. Our results demonstrate that insertion of a neomycin selection cassette into the mouse genome can have specific deleterious effects on the development of the olfactory system and highlight the importance of removing antibiotic resistance cassettes from genetic loss-of-function mouse models when studying olfactory system development.

Receptors underlying an odorant's valence across concentrations in Drosophila larvae.

Odorants interact with receptors expressed in specialized olfactory neurons, and neurons of the same class send their axons to distinct glomeruli in the brain. The stereotypic spatial glomerular activity map generates recognition and the behavioral response for the odorant. The valence of an odorant changes with concentration, typically becoming aversive at higher concentrations. Interestingly, in Drosophila larvae, the odorant (E)-2-hexenal is aversive at low concentrations and attractive at higher concentrations. We investigated the molecular and neural basis of this phenomenon, focusing on how activities of different olfactory neurons conveying opposing effects dictate behaviors. We identified the repellant neuron in the larvae as one expressing the olfactory receptor Or7a, whose activation alone at low concentrations of (E)-2-hexenal elicits an avoidance response in an Or7a-dependent manner. We demonstrate that avoidance can be overcome at higher concentrations by activation of additional neurons that are known to be attractive, most notably odorants that are known activators of Or42a and Or85c. These findings suggest that in the larval stage, the attraction-conveying neurons can overcome the aversion-conveying channels for (E)-2-hexenal.

Functional odor map heterogeneity is based on multifaceted glomerular connectivity in larval Xenopus olfactory bulb

Glomeruli are the functional units of the vertebrate olfactory bulb (OB) connecting olfactory receptor neuron (ORN) axons and mitral/tufted cell (MTC) dendrites. In amphibians, these two circuit elements regularly branch and innervate multiple, spatially distinct glomeruli. Using functional multiphoton-microscopy and single-cell tracing, we investigate the impact of this wiring on glomerular module organization and odor representations on multiple levels of the Xenopus laevis OB network. The glomerular odor map to amino acid odorants is neither stereotypic between animals nor chemotopically organized. Among the morphologically heterogeneous group of uni- and multi-glomerular MTCs, MTCs can selectively innervate glomeruli formed by axonal branches of individual ORNs. We conclude that odor map heterogeneity is caused by the coexistence of different intermingled glomerular modules. This demonstrates that organization of the amphibian main olfactory system is not strictly based on uni-glomerular connectivity.

Origin of wiring specificity in an olfactory map revealed by neuron type-specific, time-lapse imaging of dendrite targeting.

How does wiring specificity of neural maps emerge during development? Formation of the adult Drosophila olfactory glomerular map begins with the patterning of projection neuron (PN) dendrites at the early pupal stage. To better understand the origin of wiring specificity of this map, we created genetic tools to systematically characterize dendrite patterning across development at PN type-specific resolution. We find that PNs use lineage and birth order combinatorially to build the initial dendritic map. Specifically, birth order directs dendrite targeting in rotating and binary manners for PNs of the anterodorsal and lateral lineages, respectively. Two-photon- and adaptive optical lattice light-sheet microscope-based time-lapse imaging reveals that PN dendrites initiate active targeting with direction-dependent branch stabilization on the timescale of seconds. Moreover, PNs that are used in both the larval and adult olfactory circuits prune their larval-specific dendrites and re-extend new dendrites simultaneously to facilitate timely olfactory map organization. Our work highlights the power and necessity of type-specific neuronal access and time-lapse imaging in identifying wiring mechanisms that underlie complex patterns of functional neural maps.

Mapping odorant sensitivities reveals a sparse but structured representation of olfactory chemical space by sensory input to the mouse olfactory bulb.

In olfactory systems, convergence of sensory neurons onto glomeruli generates a map of odorant receptor identity. How glomerular maps relate to sensory space remains unclear. We sought to better characterize this relationship in the mouse olfactory system by defining glomeruli in terms of the odorants to which they are most sensitive. Using high-throughput odorant delivery and ultrasensitive imaging of sensory inputs, we imaged responses to 185 odorants presented at concentrations determined to activate only one or a few glomeruli across the dorsal olfactory bulb. The resulting datasets defined the tuning properties of glomeruli - and, by inference, their cognate odorant receptors - in a low-concentration regime, and yielded consensus maps of glomerular sensitivity across a wide range of chemical space. Glomeruli were extremely narrowly tuned, with ~25% responding to only one odorant, and extremely sensitive, responding to their effective odorants at sub-picomolar to nanomolar concentrations. Such narrow tuning in this concentration regime allowed for reliable functional identification of many glomeruli based on a single diagnostic odorant. At the same time, the response spectra of glomeruli responding to multiple odorants was best predicted by straightforward odorant structural features, and glomeruli sensitive to distinct odorants with common structural features were spatially clustered. These results define an underlying structure to the primary representation of sensory space by the mouse olfactory system.

Spatial transcriptomic reconstruction of the mouse olfactory glomerular map suggests principles of odor processing.

The olfactory system's ability to detect and discriminate between the vast array of chemicals present in the environment is critical for an animal's survival. In mammals, the first step of this odor processing is executed by olfactory sensory neurons, which project their axons to a stereotyped location in the olfactory bulb (OB) to form glomeruli. The stereotyped positioning of glomeruli in the OB suggests an importance for this organization in odor perception. However, because the location of only a limited subset of glomeruli has been determined, it has been challenging to determine the relationship between glomerular location and odor discrimination. Using a combination of single-cell RNA sequencing, spatial transcriptomics and machine learning, we have generated a map of most glomerular positions in the mouse OB. These observations significantly extend earlier studies and suggest an overall organizational principle in the OB that may be used by the brain to assist in odor decoding.

Main olfactory bulb reconfiguration by prolonged passive olfactory experience correlates with increased brain‐derived neurotrophic factor and improved innate olfaction

The main olfactory bulb (MOB) is highly plastic and constantly reconfiguring its function and structure depending on sensory experience. Despite the extensive evidence of anatomical, functional and behavioural changes in the olfactory system induced by highly variable olfactory experiences, it is still unknown whether prolonged passive odour experience could reconfigure the MOB at its input and network activity levels and whether these changes impact innate olfaction. Here, by measuring odour-induced glomerular activation, MOB network activity and innate olfactory behaviours, we described a profound MOB reconfiguration induced by prolonged passive olfactory experience in adult animals that impacts MOB input integration at the glomerular layer including an increase in the activated glomerular area and signal intensity, which is combined with a refinement in the number of activated glomeruli and less-overlapped glomerular maps. We also found that prolonged passive olfactory experience dramatically changes MOB population activity in the presence and absence of odours, which is reflected as a decrease in slow oscillations (<12 Hz) and an increase in fast oscillations (>12 Hz). All these functional changes in awake and anaesthetized mice correlate with an increase in brain-derived neurotrophic factor (BDNF) and with improved innate olfactory responses such as habituation/dishabituation and innate preference/avoidance. Our study shows that prolonged passive olfactory experience in adult animals produces a dramatic reconfiguration of the MOB network, possibly driven by BDNF, that improves innate olfactory responses.

The functional relevance of olfactory marker protein in the vertebrate olfactory system: a never-ending story.

Olfactory marker protein (OMP) was first described as a protein expressed in olfactory receptor neurons (ORNs) in the nasal cavity. In particular, OMP, a small cytoplasmic protein, marks mature ORNs and is also expressed in the neurons of other nasal chemosensory systems: the vomeronasal organ, the septal organ of Masera, and the Grueneberg ganglion. While its expression pattern was more easily established, OMP's function remained relatively vague. To date, most of the work to understand OMP's role has been done using mice lacking OMP. This mostly phenomenological work has shown that OMP is involved in sharpening the odorant response profile and in quickening odorant response kinetics of ORNs and that it contributes to targeting of ORN axons to the olfactory bulb to refine the glomerular response map. Increasing evidence shows that OMP acts at the early stages of olfactory transduction by modulating the kinetics of cAMP, the second messenger of olfactory transduction. However, how this occurs at a mechanistic level is not understood, and it might also not be the only mechanism underlying all the changes observed in mice lacking OMP. Recently, OMP has been detected outside the nose, including the brain and other organs. Although no obvious logic has become apparent regarding the underlying commonality between nasal and extranasal expression of OMP, a broader approach to diverse cellular systems might help unravel OMP's functions and mechanisms of action inside and outside the nose.

Visualization of antennal lobe glomeruli activated by nonappetitive D-limonene and appetitive 1-octen-3-ol odors via two types of olfactory organs in the blowfly Phormia regina

Appetite or feeding motivation relies significantly on food odors. In the blowfly Phormia regina, feeding motivation for sucrose is decreased by the odor of d-limonene but increased by the odor of 1-octen-3-ol odor. These flies have antennal lobes (ALs) consisting of several tens of glomerular pairs as a primary olfactory center in the brain. Odor information from different olfactory organs—specifically, the antennae and maxillary palps—goes to the corresponding glomeruli. To investigate how odors differently affect feeding motivation, we identified the olfactory organs and glomeruli that are activated by nonappetitive and appetitive odors. We first constructed a glomerular map of the antennal lobe in P. regina. Anterograde fluorescence labeling of antennal and maxillary afferent nerves, both of which project into the contralateral and ipsilateral ALs, revealed differential staining in glomerular regions. Some of the axonal fiber bundles from the antennae and maxillary palps projected to the subesophageal ganglion (SOG). We visualized the activation of the glomeruli in response to odor stimuli by immunostaining phosphorylated extracellular signal-regulated kinase (pERK). We observed different glomerulus activation under different odor stimulations. Referring to our glomerular map, we determined that antennal exposure to d-limonene odor activated the DA13 glomeruli, while exposure of the maxillary palps to 1-octen-3-ol activated the MxB1 glomeruli. Our results indicated that a nonappetitive odor input from the antennae and an appetitive odor input from the maxillary palps activate different glomeruli in the different regions of ALs in the blowfly P. regina. Collectively, our findings suggest that compartmentalization of glomeruli in AL is essential for proper transmission of odor information.

Simultaneous activities in both mirror-image glomerular maps in the olfactory bulb may have an important role in stress-related neuronal responses in mice

In the mouse olfactory bulb (OB), odor input from the olfactory epithelium innervates topographically to form odorant maps, which are mirror-image arrangements of glomerular clusters with domain organization. However, the functional role of the mirror-image representation in the OB remains unknown. Predator odors induce stress responses, and the dorsal domain of the dorsolateral wall of the olfactory bulb (dlOB) is known to be involved in this process. However, it remains unclear whether the activities in the medial wall of the OB (mOB), the other mirror half, are also involved in stress responses. Therefore, in this study, we investigated whether the mOB and dlOB are required for the induction of stress responses using lesioning or electrical stimulation. Although there were no significant differences in the number of activated neurons in the bed nucleus of the stria terminalis, posterior piriform cortex or amygdalo-piriform transition area, fewer activated neurons were observed in the anterior piriform cortex (APC) following lesion of both the mOB and dlOB combined. No changes were observed in the density of activated cells in any examined brain region following stimulation of either the mOB or dlOB alone. However, activated neurons in the APC were significantly more numerous following simultaneous stimulation of the mOB and dlOB. Collectively, our results suggest that simultaneous activation in both the mOB and dlOB is needed to induce APC neural activities that produce stress-like behavior. These findings provide insight into olfactory information processing, and may also help in the development of therapies for odor-induced stress behaviors.

A Prism Method for Optical Glomerular Mapping of the Medial Olfactory Bulb in Mice.

The processing of odor input in the brain begins in the olfactory bulb (OB), where odor information is represented by combinations of active glomeruli. Each glomerulus is associated with a specific odorant receptor type, of which there are ~1,000 in mice; thus different odors activate different subsets of glomeruli. Most receptor types have duplicate lateral and medial glomeruli in each of the left and right OBs. The two sets of glomeruli form separate but mirror-symmetric glomerular maps. It is not known whether the odor representations in these paired maps are exact copies of each other or potentially encode additional information. Previous studies of glomerular odor representations were mostly limited to the lateral map because the medial map is inaccessible with high-resolution activity mapping techniques, such as optical imaging. To address this, we developed a method for optical imaging of the medial bulb by replacing the contralateral bulb with a right-angle prism that has a mirror coating on the hypotenuse. With this method, we performed calcium imaging of corresponding subsets of glomeruli in the lateral map at the dorsal surface and the medial map at the medial wall. Thus, we demonstrate an experimental model system for comparing odor representations in these redundant sensory maps, enabling a better understanding of the role of paired maps and the neuronal coding of odor stimuli.

Olfactory marker protein (OMP) regulates formation and refinement of the olfactory glomerular map

Inputs from olfactory sensory neuron (OSN) axons expressing the same type of odorant receptor (OR) converge in the glomerulus of the main olfactory bulb. A key marker of mature OSNs is olfactory marker protein (OMP), whose deletion has been associated with deficits in OSN signal transduction and odor discrimination. Here, we investigate glomerular odor responses and anatomical architecture in mice in which one or both alleles of OMP are replaced by the fluorescent synaptic activity reporter, synaptopHluorin. Functionally heterogeneous glomeruli, that is, ones with microdomains with distinct odor responses, are rare in OMP+/– mice, but occur frequently in OMP–/– mice. Genetic targeting of single ORs reveals that these microdomains arise from co-innervation of individual glomeruli by OSNs expressing different ORs. This glomerular mistargeting is locally restricted to a few glomerular diameters. Our studies document functional heterogeneity in sensory input within individual glomeruli and uncover its anatomical correlate, revealing an unexpected role for OMP in the formation and refinement of the glomerular map.

Loss of odor-induced c-Fos expression of juxtaglomerular activity following maintenance of mice on fatty diets.

Diet-induced obesity (DIO) decreases the number of OMP+ olfactory sensory neurons (OSN) in the olfactory epithelium by 25% and reduces correlate axonal projections to the olfactory bulb (OB). Whether surviving OSNs have equivalent odor responsivity is largely unknown. Herein, we utilized c-fos immediate-early gene expression to map neuronal activity and determine whether mice weaned to control (CF), moderately-high fat (MHF), or high-fat (HF) diet for a period of 6months had changes in odor activation. Diet-challenged M72-IRES-tau-GFP mice were exposed to either a preferred M72 (Olfr160) ligand, isopropyl tiglate, or clean air in a custom-made Bell-jar infusion chamber using an alternating odor exposure pattern generated by a picosprizer™. Mice maintained on fatty diets weighed significantly more and cleared glucose less efficiently as determined by an intraperitoneal glucose tolerance test (IPGTT). The number of juxtaglomerular cells (JGs) decreased following maintenance of the mice on the MHF diet for cells surrounding the medial but not lateral M72 glomerulus within a 4 cell-column distance. The percentage of c-fos + JGs surrounding the lateral M72 glomerulus decreased in fat-challenged mice whereas those surrounding the medial glomerulus were not affected by diet. Altogether, these results show an asymmetry in the responsiveness of the 'mirror image' glomerular map for the M72 receptor that shows greater sensitivity of the lateral vs. medial glomerulus upon exposure to fatty diet.

A Mirror-Symmetric Excitatory Link Coordinates Odor Maps across Olfactory Bulbs and Enables Odor Perceptual Unity

Sensory input reaching the brain from bilateral and offset channels is nonetheless perceived as unified. This unity could be explained by simultaneous projections to both hemispheres, or inter-hemispheric information transfer between sensory cortical maps. Odor input, however, is not topographically organized, nor does it project bilaterally, making olfactory perceptual unity enigmatic. Here we report a circuit that interconnects mirror-symmetric isofunctional mitral/tufted cells between the mouse olfactory bulbs. Connected neurons respond to similar odors from ipsi- and contra-nostrils, whereas unconnected neurons do not respond to odors from the contralateral nostril. This connectivity is likely mediated through a one-to-one mapping from mitral/tufted neurons to the ipsilateral anterior olfactory nucleus pars externa, which activates the mirror-symmetric isofunctional mitral/tufted neurons glutamatergically. This circuit enables sharing of odor information across hemispheres in the absence of a cortical topographical organization, suggesting that olfactory glomerular maps are the equivalent of cortical sensory maps found in other senses.

Sema7A/PlxnCl signaling triggers activity-dependent olfactory synapse formation

In mammals, neural circuits are formed based on a genetic program and further refined by neuronal activity during the neonatal period. We report that in the mouse olfactory system, the glomerular map is not merely refined but newly connected to second-order neurons by odorant-receptor-derived neuronal activity. Here, we analyzed a pair of molecules, Sema7A, expressed in olfactory sensory neurons (OSNs) in an activity-dependent manner, and PlxnC1, localized to dendrites of mitral/tufted (M/T) cells in the first week after birth. In Sema7A or PlxnC1 knockout (KO) mice, initiation of synapse formation and dendrite selection of M/T cells were perturbed. Reconstitution and rescue experiments demonstrated that Sema7A–PlxnC1 interaction is essential to form the post-synaptic assembly. Pharmacological blocking experiments indicated that synaptic transmission triggers primary dendrite selection by synaptic competition. We conclude that Sema7A signaling is key to inducing activity-dependent post-synapse events and dendrite selection in M/T-cells during the neonatal period.

Differential timing of neurogenesis underlies dorsal-ventral topographic projection of olfactory sensory neurons

BackgroundThe mammalian primary olfactory system has a spatially-ordered projection in which olfactory sensory neurons (OSNs) located in the dorsomedial (DM) and ventrolateral (VL) region of the olfactory epithelium (OE) send their axons to the dorsal and ventral region of the olfactory bulb (OB), respectively. We previously found that OSN axonal projections occur sequentially, from the DM to the VL region of the OE. The differential timing of axonal projections is important for olfactory map formation because early-arriving OSN axons secrete guidance cues at the OB to help navigate late-arriving OSN axons. We hypothesized that the differential timing of axonal projections is regulated by the timing of OSN neurogenesis. To test this idea, we investigated spatiotemporal patterns of OSN neurogenesis during olfactory development.Methods and resultsTo determine the time of OSN origin, we used two thymidine analogs, BrdU and EdU, which can be incorporated into cells in the S-phase of the cell-cycle. We injected these two analogs at different developmental time points and analyzed distribution patterns of labeled OSNs. We found that OSNs with different dates of origin were differentially distributed in the OE. The majority of OSNs generated at the early stage of development were located in the DM region of the OE, whereas OSNs generated at the later stage of development were preferentially located in the VL region of the OE.ConclusionsThese results indicate that the number of OSNs is sequentially increased from the DM to the VL axis of the OE. Moreover, the temporal sequence of OSN proliferation correlates with that of axonal extension and emergence of glomerular structures in the OB. Thus, we propose that the timing of OSN neurogenesis regulates that of OSN axonal projection and thereby helps preserve the topographic order of the olfactory glomerular map along the dorsal–ventral axis of the OB.

Dense encoding of natural odorants by ensembles of sparsely activated neurons in the olfactory bulb.

Sensory information undergoes substantial transformation along sensory pathways, usually encompassing sparsening of activity. In the olfactory bulb, though natural odorants evoke dense glomerular input maps, mitral and tufted (M/T) cells tuning is considered to be sparse because of highly odor-specific firing rate change. However, experiments used to draw this conclusion were either based on recordings performed in anesthetized preparations or used monomolecular odorants presented at arbitrary concentrations. In this study, we evaluated the lifetime and population sparseness evoked by natural odorants by capturing spike temporal patterning of neuronal assemblies instead of individual M/T tonic activity. Using functional imaging and tetrode recordings in awake mice, we show that natural odorants at their native concentrations are encoded by broad assemblies of M/T cells. While reducing odorant concentrations, we observed a reduced number of activated glomeruli representations and consequently a narrowing of M/T tuning curves. We conclude that natural odorants at their native concentrations recruit M/T cells with phasic rather than tonic activity. When encoding odorants in assemblies, M/T cells carry information about a vast number of odorants (lifetime sparseness). In addition, each natural odorant activates a broad M/T cell assembly (population sparseness).