Cortical processing of auditory stimuli involves large populations of neurons with distinct individual response profiles. However, the functional organization and dynamics of local populations in the auditory cortex have remained largely unknown. Using in vivo two-photon calcium imaging, we examined the response profiles and network dynamics of layer 2/3 neurons in the primary auditory cortex (A1) of mice in response to pure tones. We found that local populations in A1 were highly heterogeneous in the large-scale tonotopic organization. Despite the spatial heterogeneity, the tendency of neurons to respond together (measured as noise correlation) was high on average. This functional organization and high levels of noise correlations are consistent with the existence of partially overlapping cortical subnetworks. Our findings may account for apparent discrepancies between ordered large-scale organization and local heterogeneity.

Summary In the mouse olfactory bulb, information from sensory neurons is extensively processed by local interneurons before being transmitted to the olfactory cortex by mitral and tufted (M/T) cells. The precise function of these local networks remains elusive because of the vast heterogeneity of interneurons, their diverse physiological properties, and their complex synaptic connectivity. Here we identified the parvalbumin interneurons (PVNs) as a prominent component of the M/T presynaptic landscape by using an improved rabies-based trans-synaptic tracing method for local circuits. In vivo two-photon targeted patch recording revealed that PVNs have exceptionally broad olfactory receptive fields, and exhibit largely excitatory and persistent odor responses. Trans-synaptic tracing indicated that PVNs receive direct input from widely distributed M/T cells. Both the anatomical and functional extent of this M/T→PVN→M/T circuit contrasts with the narrowly confined M/T→granule cell→M/T circuit, suggesting that olfactory information is processed by multiple local circuits operating at distinct spatial scales.

Motherhood is associated with different forms of physiological alterations including transient hormonal changes and brain plasticity. The underlying impact of these changes on the emergence of maternal behaviors and sensory processing within the mother's brain are largely unknown. By using in vivo cell-attached recordings in the primary auditory cortex of female mice, we discovered that exposure to pups' body odor reshapes neuronal responses to pure tones and natural auditory stimuli. This olfactory-auditory interaction appeared naturally in lactating mothers shortly after parturition and was long lasting. Naive virgins that had experience with the pups also showed an appearance of olfactory-auditory integration in A1, suggesting that multisensory integration may be experience dependent. Neurons from lactating mothers were more sensitive to sounds as compared to those from experienced mice, independent of the odor effects. These uni- and multisensory cortical changes may facilitate the detection and discrimination of pup distress calls and strengthen the bond between mothers and their neonates.

A barrage of information constantly assaults our senses, of which only a fraction is relevant at any given point in time. However, the neural circuitry supporting the suppression of irrelevant sensory distractors is not completely understood. The claustrum, a circuit hub with vast cortical connectivity, is an intriguing brain structure, whose restrictive anatomy, thin and elongated, has precluded functional investigation. Here, we describe the use of Egr2-CRE mice to access genetically defined claustral neurons. Utilizing conditional viruses for anterograde axonal labeling and retrograde trans-synaptic tracing, we validated this transgenic model for accessing the claustrum and extended the known repertoire of claustral input/output connectivity. Addressing the function of the claustrum, we inactivated CL neurons, chronically as well as acutely, in mice performing an automated two-alternative forced-choice behavioral task. Strikingly, inhibition of CL neurons did not significantly impact task performance under varying delay times and cue durations, but revealed a selective role for the claustrum in supporting performance in the presence of an irrelevant auditory distractor. Further investigation of behavior, in the naturalistic maternal pup-retrieval task, replicated the result of sensitization to an auditory distractor following inhibition of CL neurons. Initiating investigation into the underlying mechanism, we found that activation of CL neurons modulated cortical sensory processing, suppressing tone representation in the auditory cortex. This functional study, utilizing selective genetic access, implicates the claustrum in supporting resilience to distraction, a fundamental aspect of attention.

The mammalian brain maintains few developmental niches where neurogenesis persists into adulthood. One niche is located in the olfactory system where the olfactory bulb continuously receives functional interneurons. In vivo two-photon microscopy of lentivirus-labeled newborn neurons was used to directly image their development and maintenance in the olfactory bulb. Time-lapse imaging of newborn neurons over several days showed that dendritic formation is highly dynamic with distinct differences between spiny neurons and non-spiny neurons. Once incorporated into the network, adult-born neurons maintain significant levels of structural dynamics. This structural plasticity is local, cumulative and sustained in neurons several months after their integration. Thus, I provide a new experimental system for directly studying the pool of regenerating neurons in the intact mammalian brain and suggest that regenerating neurons form a cellular substrate for continuous wiring plasticity in the olfactory bulb.

Structural changes in hippocampal dendrites and dendritic spines are thought to be a consequence of a wide range of experience-and activity-dependent manipulations. We explored the dynamics of hippocampal dendritic spines in vivo by developing a surgical preparation of the adult mouse brain that enabled two-photon imaging of fluorescently labeled CA1 pyramidal neurons. Dendritic trees and spines were repeatedly visualized over many hours in exquisite detail. We tested spine stability under both control conditions and during prolonged epileptic seizures. Remarkably, spines remained structurally stable after 30 min of experimental induction of epileptic seizures. Spines began to disappear only several hours after induction of epileptic activity. We thus demonstrate that this technique provides a methodology for direct in vivo optical studies of the intact mammalian hippocampus.

The adult mammalian brain maintains a prominent stem cell niche in the subventricular zone supplying new neurons to the olfactory bulb. We examined the dynamics of synaptogenesis by imaging the formation and elimination of clusters of a postsynaptic marker (PSD95), genetically targeted to adult-born neurons. We imaged in vivo adult-born periglomerular neurons (PGNs) during two phases of development, immaturity and maturity. Immature PGNs showed high levels of PSD95 puncta dynamics during 12-72 h intervals. Mature PGNs were more stable compared with immature PGNs but still remained dynamic, suggesting that synaptogenesis persists long after these neurons integrated into the network. By combining intrinsic signal and two photon imaging we followed PSD95 puncta in sensory enriched glomeruli. Sensory input upregulated the development of adult-born PGNs only in enriched glomeruli. Our data provide evidence for an activity-based mechanism that enhances synaptogenesis of adult-born PGNs during their initial phases of development.

In many regions of the adult mammalian brain, pronounced changes in synaptic input caused by lesions or severe sensory deprivation induce marked sprouting or retraction of neuronal dendrites. In the adult olfactory bulb, adult neurogenesis produces less pronounced, but continuously ongoing synapse turnover. To test the structural stability of adult dendrites in this context, we used two-photon microscopy to image dendrites of mitral and tufted (M/T) cells over prolonged periods in adult mice. Although pharmacologically increased activity could elicit morphological changes, under natural conditions such as ongoing neurogenesis, an odor-enriched environment or olfactory-based learning, M/T cell dendrites remained highly stable. Thus, in a context of ongoing adult synaptogenesis, dendritic stability could serve as a structural scaffold to maintain the organization of local circuits.