[show abstract][hide abstract] ABSTRACT: For many mammals, individual recognition of conspecifics relies on olfactory cues. Certain individual recognition memories are thought to be stored when conspecific odor cues coincide with surges of noradrenaline (NA) triggered by intensely arousing social events. Such familiar stimuli elicit reduced behavioral responses, a change likely related to NA-dependent plasticity in the olfactory bulb (OB). In addition to its role in these ethological memories, NA signaling in the OB appears to be relevant for the discrimination of more arbitrary odorants as well. Nonetheless, no NA-gated mechanism of long-term plasticity in the OB has ever been directly observed in vivo. Here, we report that NA release from locus ceruleus (LC), when coupled to odor presentation, acts locally in the main OB to cause a specific long-lasting suppression of responses to paired odors. These effects were observed for both food odors and urine, an important social recognition cue. Moreover, in subsequent behavioral tests, mice exhibited habituation to paired urine stimuli, suggesting that this LC-mediated olfactory neural plasticity, induced under anesthesia, can store an individual recognition memory that is observable after recovery.
Journal of Neuroscience 11/2008; 28(42):10711-9. · 6.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Here we describe a knock-in mouse model for Cre-loxP-based conditional expression of TRPV1 in central nervous system neurons. Expression of Cre recombinase using biolistics, lentivirus or genetic intercrosses triggered heterologous expression of TRPV1 in a cell-specific manner. Application of the TRPV1 ligand capsaicin induced strong inward currents, triggered action potentials and activated stereotyped behaviors, allowing cell type-specific chemical genetic control of neuronal activity in vitro and in vivo.
[show abstract][hide abstract] ABSTRACT: The mammalian olfactory system recognizes an enormous variety of odorants carrying a wide range of important behavioral cues. In the main olfactory bulb (MOB), odorants are ultimately represented through the action potential activity of mitral/tufted cells (M/Ts), whose selectivity and tuning to odorant molecules are therefore fundamental determinants of MOB sensory coding. However, the sheer number and diversity of discrete olfactory stimuli has been a major barrier to comprehensively evaluating M/T selectivity. To address this issue, we assessed M/T odorant responses in anesthetized mice to a 348-odorant panel widely and systematically distributed throughout chemical space, presented both individually and in mixtures at behaviorally relevant concentrations. We found that M/T activation by odorants was markedly selective, with neurons responding robustly, sensitively, and reliably to only a highly restricted subset of stimuli. Multiple odorants activating a single neuron commonly shared clear structural similarity, but M/T tuning also frequently extended beyond obviously defined chemical categories. Cells typically responded to effective compounds presented both individually and in mixtures, although firing rates evoked by mixtures typically showed partial suppression. Response selectivity was further confirmed in awake animals by chronic recordings of M/Ts. These data indicate that individual M/Ts encode specific odorant attributes shared by only a small fraction of compounds and imply that the MOB relays the collective molecular features of an odorant stimulus through a restricted set of M/Ts, each narrowly tuned to a particular stimulus characteristic.
Journal of Neuroscience 03/2007; 27(8):2091-101. · 6.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Individual odorants activate only a small fraction of mitral cells in the mouse main olfactory bulb (MOB). Odor mixtures are represented by a combination of activated mitral cells, forming reproducible activation maps in the olfactory bulb. However, how the activation of a cohort of narrowly tuned mitral cells by odor mixtures is read out synaptically by neurons in higher-level olfactory structures, such as the anterior olfactory nucleus (AON), is mostly unknown. In the current study, we used intracellular and extracellular recordings to examine and compare responses of AON neurons and MOB mitral cells to a panel of structurally diverse odorants presented either as mixtures or as individual components. We found that a majority of individual AON neurons could be synaptically activated by several mixtures of structurally dissimilar components and by several dissimilar components in an effective mixture. The suprathreshold response of an AON neuron to an effective mixture often exceeded the sum of its suprathreshold responses to all of the components in that mixture, indicating a nonlinear combinatorial interaction. In contrast to the broad responsiveness of AON neurons, the majority of mitral cells were activated by only one or two components in a single mixture. The broader responsiveness of AON neurons relative to mitral cells suggests that individual AON neurons synaptically integrate several functionally distinct mitral cell inputs.
Journal of Neuroscience 12/2006; 26(46):12023-32. · 6.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Natural odorants are complex mixtures of diverse chemical compounds. Monomolecular odorants are represented in the main olfactory bulb by distinct spatial patterns of activated glomeruli. However, it remains unclear how individual compounds contribute to population representations of natural stimuli, which appear to be unexpectedly sparse. We combined gas chromatography and intrinsic signal imaging to visualize glomerular responses to natural stimuli and their fractionated components. While whole stimuli activated up to 20 visible glomeruli, each fractionated component activated only one or few glomeruli, and most glomeruli were activated by only one component. Thus, responses to complex mixtures reflected activation by multiple components, with each contributing only a small part of the overall representation. We conclude that the population response to a complex stimulus is largely the sum of the responses to its individual components, and activation of an individual glomerulus independently signals the presence of a specific component.
[show abstract][hide abstract] ABSTRACT: As a consequence of adult neurogenesis, the olfactory bulb (OB) receives a continuous influx of newborn neurons well into adulthood. However, their rates of generation and turnover, the factors controlling their survival, and how newborn neurons intercalate into adult circuits are largely unknown. To visualize the dynamics of adult neurogenesis, we produced a line of transgenic mice expressing GFP in approximately 70% of juxtaglomerular neurons (JGNs), a population that undergoes adult neurogenesis. Using in vivo two-photon microscopy, time-lapse analysis of identified JGN cell bodies revealed a neuronal turnover rate of approximately 3% of this population per month. Although new neurons appeared and older ones disappeared, the overall number of JGNs remained constant. This approach provides a dynamic view of the actual appearance and disappearance of newborn neurons in the vertebrate central nervous system, and provides an experimental substrate for functional analysis of adult neurogenesis.
Proceedings of the National Academy of Sciences 03/2006; 103(6):1912-7. · 9.74 Impact Factor
[show abstract][hide abstract] ABSTRACT: Mammalian urine releases complex mixtures of volatile compounds that are used in reproduction, territoriality and conspecific recognition. To understand how such complex mixtures are represented in the main olfactory bulb, we analysed the electrophysiological responses of individual mitral cells to volatile compounds in mouse urine. In both males and females, urine volatile compounds evoke robust responses in a small subset of mitral cells. Fractionation of the volatile compounds using gas chromatography showed that out of the hundreds of compounds present, mitral cells are activated by single compounds. One cohort of mitral cells responded exclusively to male urine; these neurons were activated by (methylthio)methanethiol, a potent, previously unknown semiochemical present only in male urine. When added to urine, synthetic (methylthio)methanethiol significantly enhances urine attractiveness to female mice. We conclude that mitral cells represent natural odorant stimuli by acting as selective feature detectors, and that their activation is largely independent of the presence of other components in the olfactory stimulus.
[show abstract][hide abstract] ABSTRACT: The visual system encodes and deciphers information using parallel, anatomically segregated, processing streams. To reveal patterns of gene expression in the visual thalamus correlated with physiological processing streams, we designed a custom ferret cDNA microarray. By isolating specific subregions and layers of the thalamus, we identified a set of transcription factors, including Zic2, Islet1, and Six3, the unique distribution profiles of which differentiated the lateral geniculate nucleus (LGN) from the associated perigeniculate nucleus. Within the LGN, odd homeobox1 differentiated the A layers, which contain X cells and Y cells, from the C layers. One neuron-specific protein, Purkinje cell protein 4 (PCP4), was strongly expressed in Y cells in the ferret LGN and in the magnocellular layers of the primate LGN. In the ferret LGN, PCP4 expression began as early as postnatal day 7 (P7), suggesting that Y cells are already specified by P7. These results reveal a rich molecular repertoire that correlates with functional divisions of the LGN.
Journal of Neuroscience 12/2004; 24(44):9962-70. · 6.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Odorant receptors (ORs) form one of the largest gene families in the genome. However, the vast majority are orphan receptors as the ligands that activate them remain unknown. Deorphaning approaches have generally focused on finding ligands for particular receptors expressed in homologous or heterologous cells; these attempts have met with only partial success. Here, we outline a conceptually different strategy in which we search for odorant receptors activated by a known odorant. Intrinsic signal imaging of the main olfactory bulb is first used to locate activated glomeruli in vivo, followed by retrograde tracing to label the sensory neurons in the olfactory epithelium projecting to the activated glomerulus. Subsequently, single cell RT-PCR is used to reveal the identity of the odorant receptors expressed in retrogradely labeled neurons. To demonstrate the applicability of this method, we searched for candidate ORs responding to the aldehyde odorant butanal. This method may be a useful tool to decipher specific ligand--OR interactions in the mouse olfactory bulb.
[show abstract][hide abstract] ABSTRACT: The past few years have delivered substantial progress in understanding the molecular logic of the mammalian vomeronasal system. Selective expression of vomeronasal receptors and high response selectivity of vomeronasal receptor neurons suggest that pheromones are encoded by labeled lines at the level of the vomeronasal organ: each pheromonal compound is represented by the activation of a small and exclusive subset of receptor neurons. Labeled lines might be transferred to the accessory olfactory bulb through convergent connections. The key challenges ahead will be to identify the pheromonal ligands of the receptors and unravel the functional connectivity from the vomeronasal organ to the hypothalamus.
Current Opinion in Neurobiology 09/2004; 14(4):428-34. · 7.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: 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.
Journal of Neuroscience 04/2004; 24(13):3147-51. · 6.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: 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.
[show abstract][hide abstract] ABSTRACT: In rodents, each main olfactory bulb contains two mirror-symmetric glomerular maps, a feature not found in the initial topographic maps of other sensory systems. Targeting tracer injections to identified glomeruli revealed that isofunctional odor columns-translaminar assemblies connected to a given glomerulus-were specifically and reciprocally interconnected through a mutually inhibitory circuit with exquisite topographic specificity. Thus, instead of containing two mirror-symmetric maps, we propose that the olfactory bulb contains a single integrated map in which isofunctional odor columns are connected through an intrabulbar link, analogous to the specific horizontal connections linking iso-orientation columns in primary visual cortex.
[show abstract][hide abstract] ABSTRACT: Many mammalian species rely on pheromones-semiochemicals produced by other members of the same species-to communicate social status and reproductive readiness. To assess how the central nervous system integrates the complex repertoire of pheromones, we recorded from single neurons in the accessory olfactory bulb, a nucleus that processes pheromonal signals, of male mice engaged in natural behaviors. Neuronal firing was robustly modulated by physical contact with male and female conspecifics, with individual neurons activated selectively by specific combinations of the sex and strain of conspecifics. We infer that mammals encode social and reproductive information by integrating vomeronasal sensory activity specific to sex and genetic makeup.
[show abstract][hide abstract] ABSTRACT: The horizontal network in visual cortex layer 2/3 is implicated in numerous psychophysical and physiological properties. To investigate the spatial and temporal distribution of excitation and inhibition evoked by this network, we used voltage-sensitive dyes to image the responses to focal electrical stimulation in tangential slices of ferret visual cortex layer 2/3. The resulting optical patterns included a diffuse zone of activation near the stimulation site and numerous ovoid domains throughout the slice. In contrast to the fixed anatomy of the horizontal connections, substantial shifts in both space and time were evident in the distribution of population-based neuronal activity during stimulus trains. Both of these shifts relied on inhibitory synaptic potentials, suggesting that inhibition driven by horizontal connections sculpts the distribution of activity in this cortical network.
Journal of Neurophysiology 02/2003; 89(1):488-500. · 3.30 Impact Factor
[show abstract][hide abstract] ABSTRACT: To investigate how neurons in cortical layer 2/3 integrate horizontal inputs arising from widely distributed sites, we combined intracellular recording and voltage-sensitive dye imaging to visualize the spatiotemporal dynamics of neuronal activity evoked by electrical stimulation of multiple sites in visual cortex. Individual stimuli evoked characteristic patterns of optical activity, while delivering stimuli at multiple sites generated interacting patterns in the regions of overlap. We observed that neurons in overlapping regions received convergent horizontal activation that generated nonlinear responses due to the emergence of large inhibitory potentials. The results indicate that co-activation of multiple sets of horizontal connections recruit strong inhibition from local inhibitory networks, causing marked deviations from simple linear integration.
Journal of Neurophysiology 02/2003; 89(1):501-12. · 3.30 Impact Factor
[show abstract][hide abstract] ABSTRACT: In cultured neurons, the exogenous application of neurotrophins (in homogenous concentrations) alters many features of axonal and dendritic arbors. In vivo, however, release of endogenous neurotrophins from neuronal processes creates spatially heterogeneous neurotrophin distributions. To probe the consequences of such endogenous neurotrophin distribution, we produced 'donor neurons' in ferret cortex brain slices that co-expressed brain-derived neurotrophic factor (BDNF) and red fluorescent protein (RFP). Using two-photon microscopy, we analyzed their effects on 'recipient neurons' that expressed green fluorescent protein (GFP) alone. BDNF released from dendrites and cell bodies acted directly on nearby recipient neurons to increase dendritic branching in a distance-dependent manner. Three-dimensional analysis of donor and recipient dendrites indicated that the BDNF source had to be within 4.5 microm to induce dendritic growth in the recipient neuron. Thus, BDNF released from an individual cell alters the structure of nearby dendrites on an exquisitely local scale.
[show abstract][hide abstract] ABSTRACT: The mammalian olfactory system detects and discriminates thousands of odorants using many different receptors expressed by sensory neurons in the nasal epithelium. Axonal projections from these neurons to the main olfactory bulbs form reproducible patterns of glomeruli in two widely separated regions of each bulb, creating two mirror-symmetric maps of odorant receptor projections. To investigate whether odorant receptors organize neural circuitry in the olfactory bulb, we have examined a genetically modified mouse line, rI7 --> M71, in which a functionally characterized receptor, rI7, has been substituted into the M71 receptor locus. Here we show that despite their ectopic location the resulting glomeruli are responsive to known ligands of the rI7 receptor, attract postsynaptic innervation by mitral/tufted cell dendrites, and endow these cells with responses that are characteristic of the rI7 receptor. External tufted cells receiving input from rI7 --> M71 glomeruli form precise intrabulbar projections that link medial and lateral rI7 --> M71 glomeruli anatomically, thus providing a substrate for coordinating isofunctional glomeruli. We conclude that odorant receptor identity in epithelial neurons determines not only glomerular convergence and function, but also functional circuitry in the olfactory bulb.
[show abstract][hide abstract] ABSTRACT: New approaches to the study of ocular dominance development, a model system for the development of neural architecture, indicate that eye-specific columns in primary visual cortex emerge substantially before the onset of the critical period, during which neural connections can be altered by visual experience. The timing, speed and specificity of column emergence implicate molecular patterning mechanisms, along with patterns of neural activity, in the generation of this columnar architecture.
Current Opinion in Neurobiology 03/2002; 12(1):104-9. · 7.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: The development of ocular dominance columns has served as a Rosetta stone for understanding the mechanisms that guide the construction of cortical circuits. Traditionally, the emergence of ocular dominance columns was thought to be closely tied to the critical period, during which columnar architecture is highly susceptible to alterations in visual input. However, recent findings in cats, monkeys and ferrets indicate that columns develop far earlier, more rapidly and with considerably greater precision than was previously suspected. These observations indicate that the initial establishment of cortical functional architecture, and its subsequent plasticity during the critical period, are distinct developmental phases that might reflect distinct mechanisms.