Spatial Structure of Synchronized Inhibition in the Olfactory Bulb.

Olfactory sensory input is detected by receptor neurons in the nose, which then send information to the olfactory bulb (OB), the first brain region for processing olfactory information. Within the OB, many local circuit interneurons, including axonless granule cells, function to facilitate fine odor discrimination. How interneurons interact with principal cells to affect bulbar processing is not known, but the mechanism is likely to be different from that in sensory cortical regions because the OB lacks an obvious topographical organization. Neighboring glomerular columns, representing inputs from different receptor neuron subtypes, typically have different odor tuning. Determining the spatial scale over which interneurons such as granule cells can affect principal cells is a critical step toward understanding how the OB operates. We addressed this question by assaying inhibitory synchrony using intracellular recordings from pairs of principal cells with different intersomatic spacing. We found, in acute rat OB slices from both sexes, that inhibitory synchrony is evident in the spontaneous synaptic input in mitral cells (MCs) separated up to 220 µm (300 µm with elevated K+). At all intersomatic spacing assayed, inhibitory synchrony was dependent on Na+ channels, suggesting that action potentials in granule cells function to coordinate GABA release at relatively distant dendrodendritic synapses formed throughout the dendritic arbor. Our results suggest that individual granule cells are able to influence relatively large groups of MCs and tufted cells belonging to clusters of at least 15 glomerular modules, providing a potential mechanism to integrate signals reflecting a wide variety of odorants.SIGNIFICANCE STATEMENT Inhibitory circuits in the olfactory bulb (OB) play a major role in odor processing, especially during fine odor discrimination. However, how inhibitory networks enhance olfactory function, and over what spatial scale they operate, is not known. Interneurons are potentially able to function on both a highly localized, synapse-specific level and on a larger, spatial scale that encompasses many different glomerular channels. Although recent indirect evidence has suggested a relatively localized functional role for most inhibition in the OB, in the present study, we used paired intracellular recordings to demonstrate directly that inhibitory local circuits operate over large spatial scales by using fast action potentials to link GABA release at many different synaptic contacts formed with principal cells.

Cell-type-specific repression by methyl-CpG-binding protein 2 is biased toward long genes.

Mutations in methyl-CpG-binding protein 2 (MeCP2) cause Rett syndrome and related autism spectrum disorders (Amir et al., 1999). MeCP2 is believed to be required for proper regulation of brain gene expression, but prior microarray studies in Mecp2 knock-out mice using brain tissue homogenates have revealed only subtle changes in gene expression (Tudor et al., 2002; Nuber et al., 2005; Jordan et al., 2007; Chahrour et al., 2008). Here, by profiling discrete subtypes of neurons we uncovered more dramatic effects of MeCP2 on gene expression, overcoming the "dilution problem" associated with assaying homogenates of complex tissues. The results reveal misregulation of genes involved in neuronal connectivity and communication. Importantly, genes upregulated following loss of MeCP2 are biased toward longer genes but this is not true for downregulated genes, suggesting MeCP2 may selectively repress long genes. Because genes involved in neuronal connectivity and communication, such as cell adhesion and cell-cell signaling genes, are enriched among longer genes, their misregulation following loss of MeCP2 suggests a possible etiology for altered circuit function in Rett syndrome.

Robust encoding of stimulus identity and concentration in the accessory olfactory system.

Sensory systems represent stimulus identity and intensity, but in the neural periphery these two variables are typically intertwined. Moreover, stable detection may be complicated by environmental uncertainty; stimulus properties can differ over time and circumstance in ways that are not necessarily biologically relevant. We explored these issues in the context of the mouse accessory olfactory system, which specializes in detection of chemical social cues and infers myriad aspects of the identity and physiological state of conspecifics from complex mixtures, such as urine. Using mixtures of sulfated steroids, key constituents of urine, we found that spiking responses of individual vomeronasal sensory neurons encode both individual compounds and mixtures in a manner consistent with a simple model of receptor-ligand interactions. Although typical neurons did not accurately encode concentration over a large dynamic range, from population activity it was possible to reliably estimate the log-concentration of pure compounds over several orders of magnitude. For binary mixtures, simple models failed to accurately segment the individual components, largely because of the prevalence of neurons responsive to both components. By accounting for such overlaps during model tuning, we show that, from neuronal firing, one can accurately estimate log-concentration of both components, even when tested across widely varying concentrations. With this foundation, the difference of logarithms, log A - log B = log A/B, provides a natural mechanism to accurately estimate concentration ratios. Thus, we show that a biophysically plausible circuit model can reconstruct concentration ratios from observed neuronal firing, representing a powerful mechanism to separate stimulus identity from absolute concentration.

Chemosensory burst coding by mouse vomeronasal sensory neurons.

The capabilities of any sensory system are ultimately constrained by the properties of the sensory neurons: the ability to detect and represent stimuli is limited by noise due to spontaneous activity, and optimal decoding in downstream circuitry must be matched to the nature of the encoding performed at the input. Here, we investigated the firing properties of sensory neurons in the accessory olfactory system, a distinct sensory system specialized for detection of socially relevant odors. Using multielectrode array recording, we observed that sensory neurons are spontaneously active and highly variable across time and trials and that this spontaneous activity limits the ability to distinguish sensory responses from noise. Sensory neuron activity tended to consist of bursts that maintained remarkably consistent statistics during both spontaneous activity and in response to stimulation with sulfated steroids. This, combined with pharmacological and genetic intervention in the signal transduction cascade, indicates that sensory transduction plays a role in shaping overall spontaneous activity. These findings indicate that as-yet unexplored characteristics of the sensory transduction cascade significantly constrain the representation of sensory information by vomeronasal neurons.

Representation and transformation of sensory information in the mouse accessory olfactory system.

In mice, nonvolatile social cues are detected and analyzed by the accessory olfactory system (AOS). Here we provide a first view of information processing in the AOS with respect to individual chemical cues. 12 sulfated steroids, recently discovered mouse AOS ligands, caused widespread activity among vomeronasal sensory neurons (VSNs), yet VSN responses clustered into a small number of repeated functional patterns or processing streams. Downstream neurons in the accessory olfactory bulb (AOB) responded to these ligands with enhanced signal/noise compared to VSNs. Although the dendritic connectivity of AOB mitral cells suggests the capacity for broad integration, most sulfated steroid responses were well-modeled by linear excitatory drive from just one VSN processing stream. However, a substantial minority demonstrated multi-stream integration. Most VSN excitation patterns were also observed in the AOB, but excitation by estradiol sulfate processing streams was rare, suggesting AOB circuit organization is specific to the biological relevance of sensed cues.

Multielectrode array recordings of the vomeronasal epithelium.

Understanding neural circuits requires methods to record from many neurons simultaneously. For in vitro studies, one currently available technology is planar multielectrode array (MEA) recording. Here we document the use of MEAs to study the mouse vomeronasal organ (VNO), which plays an essential role in the detection of pheromones and social cues via a diverse population of sensory neurons expressing hundreds of types of receptors. Combining MEA recording with a robotic liquid handler to deliver chemical stimuli, the sensory responses of a large and diverse population of neurons can be recorded. The preparation allows us to remove the intact neuroepithelium of the VNO from the mouse and stimulate with a battery of chemicals or potential ligands while monitoring the electrical activity of the neurons for several hours. Therefore, this technique serves as a useful method for assessing ligand activity as well as exploring the properties of receptor neurons. We present the techniques needed to prepare the vomeronasal epithelium, MEA recording, and chemical stimulation.