Single Neuronal Population Research Articles

FACS-array profiling of striatal projection neuron subtypes in juvenile and adult mouse brains

A major challenge in systems neuroscience is to perform precise molecular genetic analyses of a single neuronal population in the context of the complex mammalian brain. Existing technologies for profiling cell type-specific gene expression are largely limited to immature or morphologically identifiable neurons. In this study, we developed a simple method using fluorescent activated cell sorting (FACS) to purify genetically labeled neurons from juvenile and adult mouse brains for gene expression profiling. We identify and verify a new set of differentially expressed genes in the striatonigral and striatopallidal neurons, two functionally and clinically important projection neuron subtypes in the basal ganglia. We further demonstrate that Ebf1 is a lineage-specific transcription factor essential to the differentiation of striatonigral neurons. Our study provides a general approach for profiling cell type-specific gene expression in the mature mammalian brain and identifies a set of genes critical to the function and dysfunction of the striatal projection neuron circuit.

Cumulative Activation of Akt and Consequent Inhibition of Glycogen Synthase Kinase-3 by Brain-Derived Neurotrophic Factor and Insulin-Like Growth Factor-1 in Cultured Hippocampal Neurons

Brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-1 (IGF-1) seem to play key roles in mediating neuronal plasticity in the hippocampus. In the current studies, we have used cultured hippocampal neurons to study possible interactions between the two growth factors in modulating neuronal signaling pathways. BDNF and IGF-1 were found to each effectively activate the neuroprotective Akt pathway, with the magnitude of activation being at least additive when cultures were simultaneously treated with supramaximal concentrations of peptides. Likewise, a cumulative inhibitory Akt-dependent phosphorylation of proapoptotic glycogen synthase kinase-3 was observed. Immunofluorescent studies demonstrated that a single population of neurons responded to BDNF and IGF-1. In contrast, the magnitude of BDNF-stimulated extracellular signal-regulated kinase (ERK) activation was found to be much greater than that of IGF-1-stimulated ERK, such that the difference in magnitude stimulated by BDNF in the presence and absence of IGF-1 did not reach statistical significance. Consistent with the observed agonist-stimulated activation of Akt, BDNF and IGF-1 were both found to act as neurotrophins, enhancing neuronal survival under low-insulin culture conditions. Maximal survival was achieved when both growth factors were present. These findings provide insight into the significance of multiple growth factors stimulating activation of ERK and Akt in the central nervous system. In some cases, the magnitude of activation required to elicit biological responses may be achieved only with a combination of compounds.

Fiber connections of the central nucleus of semicircular torus in cyprinids

Fiber connections of the presumed auditory portion of the semicircular torus or the central nucleus (TSc) as well as other central and peripheral auditory structures were studied by tract-tracing methods in carp and goldfish. Major ascending projections to the TSc originated from the descending octaval nucleus (DO) and medullary secondary octaval population (SO). Toropetal neurons of the DO were within the saccular terminal zone. A small number of toropetal DO neurons might receive inputs from other inner ear and lateral line endorgans as well. Fibers from the DO terminated in a deep zone of the TSc, while those from the SO in both deep and superficial zones. Afferents to the TSc also arose from the rhombencephalic reticular formation, anterior octaval nucleus, isthmic reticular nucleus, perilemniscular nucleus, medial pretoral nucleus, anterior tuberal nucleus, and central posterior thalamic nucleus. The TSc projected ascending fibers to the medial pretoral nucleus, anterior tuberal nucleus, central posterior thalamic nucleus, and the preglomerular complex (anterior preglomerular nucleus, the caudomedial region of lateral preglomerular nucleus, and a medial zone of medial preglomerular nucleus). These parts of preglomerular complex appear structurally continuous sharing common hodological and cytoarchitectonic features, and thus might be regarded as a single neuronal population. Abundant descending pathways were also noted in the present study. Of particular note is the medial pretoral nucleus, which received fibers from diencephalic auditory nuclei. The nucleus gave rise to indirect descending pathways to a medial zone of the cerebellar crest where dendrites of crest cells in the SO ramify.

Propagator theory of brain dynamics

A physiologically based continuum model of brain dynamics is extended to incorporate arbitrary numbers of structures and neural populations, multiple outgoing fields of activity from a single population of neurons to various targets, improved treatment of converging or diverging projections and mesoscopic structure, and generalized connections to quantities observable via electroencephalography and other methods. The results are applied to study the corticothalamic system, predicting an intracortical resonance that leads to enhancements of electroencephalographic activity in the gamma (>30 Hz) range. This resonance involves feedback loops incorporating slow, short-range inhibitory fibers.

Background-activity-dependent properties of a network model for working memory that incorporates cellular bistability

In models of working memory, transient stimuli are encoded by feature-selective persistent neural activity. Network models of working memory are also implicitly bistable. In the absence of a brief stimulus, only spontaneous, low-level, and presumably nonpatterned neural activity is seen. In many working-memory models, local recurrent excitation combined with long-range inhibition (Mexican hat coupling) can result in a network-induced, spatially localized persistent activity or "bump state" that coexists with a stable uniform state. There is now renewed interest in the concept that individual neurons might have some intrinsic ability to sustain persistent activity without recurrent network interactions. A recent visuospatial working-memory model (Camperi and Wang 1998) incorporates both intrinsic bistability of individual neurons within a firing rate network model and a single population of neurons on a ring with lateral inhibitory coupling. We have explored this model in more detail and have characterized the response properties with changes in background synaptic input I(o) and stimulus width. We find that only a small range of I(o) yields a working-memory-like coexistence of bump and uniform solutions that are both stable. There is a rather larger range where only the bump solution is stable that might correspond instead to a feature-selective long-term memory. Such a network therefore requires careful tuning to exhibit working-memory-like function. Interestingly, where bumps and uniform stable states coexist, we find a continuous family of stable bumps representing stimulus width. Thus, in the range of parameters corresponding to working memory, the model is capable of capturing a two-parameter family of stimulus features including both orientation and width.

Correlation between multiple climbing fibre regression and parallel fibre response development in the postnatal mouse cerebellum

At the neuromuscular junction elimination of supernumerary synaptic connections during development is owing to a competitive process between single neuronal populations, whereas in the central nervous system the interaction of different types of input could affect the process. In the cerebellum, the regression from multiple- to mono-innervation of the Purkinje cells by climbing fibres is virtually completed during the first two weeks of postnatal development. While it is clear that parallel fibres are important in the control of the regression, there are conflicting results in relation to whether an early phase of regression is independent of parallel fibre effects. We studied the precise timing of climbing fibre synapse development and decline and the relationship with the functional maturation of parallel fibres. Until postnatal day (P) 6 or 7, the synaptic currents generated by different climbing fibres become progressively more uniform in amplitude. However, between P7 and 14, the amplitudes of the currents increasingly diverge until only one fibre remains connected. These data are taken as evidence that, in multiply innervated Purkinje cells, competition between different climbing fibres appears at P7 and continues during the second postnatal week. Morphological and electrophysiological data demonstrate that parallel fibres synapses appear at P7 and their development is significantly correlated with the time course of the climbing fibre regression. These results provide no evidence for climbing fibre regression independent of parallel fibres before P7 and also suggest a dominant role of the parallel fibre input in the later phase.

Neuronal Subtype-Specific Genes that Control Corticospinal Motor Neuron Development In Vivo

Within the vertebrate nervous system, the presence of many different lineages of neurons and glia complicates the molecular characterization of single neuronal populations. In order to elucidate molecular mechanisms underlying the specification and development of corticospinal motor neurons (CSMN), we purified CSMN at distinct stages of development in vivo and compared their gene expression to two other pure populations of cortical projection neurons: callosal projection neurons and corticotectal projection neurons. We found genes that are potentially instructive for CSMN development, as well as genes that are excluded from CSMN and are restricted to other populations of neurons, even within the same cortical layer. Loss-of-function experiments in null mutant mice for Ctip2 (also known as Bcl11b), one of the newly characterized genes, demonstrate that it plays a critical role in the development of CSMN axonal projections to the spinal cord in vivo, confirming that we identified central genetic determinants of the CSMN population.

A single population of olfactory sensory neurons mediates an innate avoidance behaviour in Drosophila.

All animals exhibit innate behaviours in response to specific sensory stimuli that are likely to result from the activation of developmentally programmed neural circuits. Here we observe that Drosophila exhibit robust avoidance to odours released by stressed flies. Gas chromatography and mass spectrometry identifies one component of this 'Drosophila stress odorant (dSO)' as CO2. CO2 elicits avoidance behaviour, at levels as low as 0.1%. We used two-photon imaging with the Ca2+-sensitive fluorescent protein G-CaMP to map the primary sensory neurons governing avoidance to CO2. CO2 activates only a single glomerulus in the antennal lobe, the V glomerulus; moreover, this glomerulus is not activated by any of 26 other odorants tested. Inhibition of synaptic transmission in sensory neurons that innervate the V glomerulus, using a temperature-sensitive Shibire gene (Shi(ts)), blocks the avoidance response to CO2. Inhibition of synaptic release in the vast majority of other olfactory receptor neurons has no effect on this behaviour. These data demonstrate that the activation of a single population of sensory neurons innervating one glomerulus is responsible for an innate avoidance behaviour in Drosophila.

Intrinsic bursting activity in the pre-Bötzinger complex: role of persistent sodium and potassium currents.

Computational models of single pacemaker neuron and neural population in the pre-Bötzinger Complex (pBC) were developed based on the previous models by Butera et al. (1999a,b). Our modeling study focused on the conditions that could define endogenous bursting vs. tonic activity in single pacemaker neurons and population bursting vs. asynchronous firing in populations of pacemaker neurons. We show that both bursting activity in single pacemaker neurons and population bursting activity may be released or suppressed depending on the expression of persistent sodium ( I(Na) P) and delayed-rectifier potassium ( I(K)) currents. Specifically, a transition from asynchronous firing to population bursting could be induced by a reduction of I(K) via a direct suppression of the potassium conductance or through an elevation of extracellular potassium concentration. Similar population bursting activity could be triggered by an augmentation of I(Na) P. These findings are discussed in the context of the possible role of population bursting activity in the pBC in the respiratory rhythm generation in vivo vs. in vitro and during normal breathing in vivo vs. gasping.

Orbitofrontal cortex: neuronal representation of oral temperature and capsaicin in addition to taste and texture

The primate orbitofrontal cortex is a site of convergence of information from primary taste, olfactory and somatosensory cortical areas. We describe the discovery of a population of single neurons in the macaque orbitofrontal cortex that responds to the temperature of a liquid in the mouth. The temperature stimuli consisted of water at 10 °C, 23 °C, 37 °C and 42 °C. Twenty-six of the 1149 neurons analyzed (2.3%) responded to oral temperature. The tuning profiles of the neurons to temperature showed that some of the neurons had graded responses to increasing temperature (27%), others responded to cold (10 °C) stimuli (27%), and others were tuned to temperature (46%). The neuronal responses were also measured to taste stimuli, viscosity stimuli (carboxymethyl-cellulose in the range 1–10,000 cP), and capsaicin (10 μM). Of 70 neurons with responses to any of these stimuli, 7.1% were unimodal temperature; 11.3% were temperature and taste-sensitive; 7.1% were temperature and viscosity-sensitive; and 11.3% were temperature, taste and viscosity sensitive. Capsaicin activated 15.7% of the population of responsive neurons tested. These results provide the first evidence of how the temperature of what is in the mouth is represented at the neuronal level in the orbitofrontal cortex and the first evidence for any primate cortical area that in some cases this information converges onto single neurons with inputs produced by other sensory properties of food, including taste and texture. The results provide a basis for understanding how particular combinations of oral temperature, taste, and texture can influence the palatability of foods.

Representations of the texture of food in the primate orbitofrontal cortex: neurons responding to viscosity, grittiness, and capsaicin.

The primate orbitofrontal cortex (OFC) is a site of convergence from taste, olfactory, and somatosensory cortical areas. We describe a population of single neurons in the macaque OFC that responds to the texture of food in the mouth. Use of oral viscosity stimuli consisting of carboxymethylcellulose (CMC) in the range 1-10,000 centipoise showed that the responses of one subset of these neurons were related to stimulus viscosity. Some of the neurons had increasing responses to increasing viscosity, some had decreasing responses, and some neurons were tuned to a range of viscosities. These neurons are a different population to oral fat-sensitive neurons, in that their responses to fats (e.g., safflower oil), to silicone oil [(Si(CH3)2O)n], and to mineral oil (hydrocarbon) depended on the viscosity of these oils. Thus there is a dissociation between texture channels used to sense viscosity and fat. Some of these viscosity-sensitive single neurons were unimodal (somatosensory; 25%) and some received convergent taste inputs (75%). A second subpopulation of neurons responded to gritty texture (produced by microspheres suspended in CMC). A third subpopulation of neurons responded to capsaicin. These results provide evidence about the information channels used to represent the texture and flavor of food in a part of the brain important in appetitive responses to food and are relevant to understanding the physiological and pathophysiological processes related to food intake, food selection, and the effects of variety of food texture in combination with taste and other inputs that affect food intake.

Neurons in the primate orbitofrontal cortex respond to fat texture independently of viscosity.

The primate orbitofrontal cortex (OFC) is a site of convergence from primary taste, olfactory, and somatosensory cortical areas. We describe the responses of a population of single neurons in the OFC that respond to orally applied fat (e.g., safflower oil) and to substances with a similar texture but different chemical composition, such as mineral oil (hydrocarbon) and silicone oil [(Si(CH3)2O)n]. These findings provide evidence that the neurons respond to the oral texture of fat, sensed by the somatosensory system. Use of an oral viscosity stimulus consisting of carboxymethyl-cellulose in the range 1-10,000 centipoise (cP) showed that the responses of these fat-sensitive neurons are not related to stimulus viscosity. Thus a textural component independent of viscosity and related to the slick or oily property is being used to activate these oral fat-sensitive neurons. Moreover, a separate population of neurons responds to viscosity (produced, e.g., by the carboxymethyl-cellulose series), but not to fat with the same viscosity. Thus there is a dissociation between texture channels used to sense fat viscosity and non-fat-produced viscosity. Further, free fatty acids such as linoleic acid do not activate these neurons, providing further evidence that the oral fat-sensing mechanism through which these OFC neurons are activated is not gustatory but textural. Most of this population of fat-sensitive neurons receive convergent taste inputs. These results provide evidence about how oral fat is sensed and are relevant to understanding the physiological and pathophysiological processes related to fat intake.

Sensory cortical dynamics

Sensory cortical networks are commonly regarded as stable, changing only in the face of prolonged alteration of sensory input. There is increasing evidence, however, that the functional connectivity of cortical networks changes significantly, but reversibly, in response to conditions of sensory stimulation similar to those encountered in everyday life. In this review, we provide examples of sensory cortical dynamics at the single neuron and neural population levels. The dynamics detected at both levels of experimental observation suggest that a brief exposure (tens of milliseconds to tens of seconds) to sensory stimulation is accompanied by changes in the capacity of cortical networks to process and represent environmental stimuli. Candidate cellular mechanisms and the potential benefits of such stimulus-driven, rapid, and fully reversible sensory cortical dynamics are discussed.

Activity-dependent development of axonal and dendritic delays, or, why synaptic transmission should be unreliable.

Systematic temporal relations between single neuronal activities or population activities are ubiquitous in the brain. No experimental evidence, however, exists for a direct modification of neuronal delays during Hebbian-type stimulation protocols. We show that in fact an explicit delay adaptation is not needed if one assumes that the synaptic strengths are modified according to the recently observed temporally asymmetric learning rule with the downregulating branch dominating the upregulating branch. During development, slow, unbiased fluctuations in the transmission time, together with temporally correlated network activity, may control neural growth and implicitly induce drifts in the axonal delays and dendritic latencies. These delays and latencies become optimally tuned in the sense that the synaptic response tends to peak in the soma of the postsynaptic cell if this is most likely to fire. The nature of the selection process requires unreliable synapses in order to give successful synapses an evolutionary advantage over the others. The width of the learning function also determines the preferred dendritic delay and the preferred width of the postsynaptic response. Hence, it may implicitly determine whether a synaptic connection provides a precisely timed or a broadly tuned "contextual" signal.

Single neuron burst firing in the human hippocampus during sleep.

Although there are numerous non-primate studies of the single neuron correlates of sleep-related hippocampal EEG patterns, very limited hippocampal neuronal data are available for correlation with human sleep. We recorded human hippocampal single neuron activity in subjects implanted with depth electrodes required for medical diagnosis and quantitatively evaluated discharge activity from each neuron during episodes of wakefulness (Aw), combined stage 3 and 4 slow-wave sleep (SWS), and rapid eye movement (REM) sleep. The mean firing rate of the population of single neurons was significantly higher during SWS and Aw compared with REM sleep (p = 0.002; p < 0.0001). In addition, burst firing was significantly greater during SWS compared with Aw (p = 0.001) and REM sleep (p < 0.0001). The synchronized state of SWS and associated high-frequency burst discharge found in human hippocampus may subserve functions similar to those reported in non-primate hippocampus that require burst firing to induce synaptic modifications in hippocampal circuitry and in hippocampal projections to neocortical targets that participate in memory consolidation.

Role of glutamate delta -2 receptors in activity-dependent competition between heterologous afferent fibers.

A principle that regulates detailed architecture in the brain is that active terminals have a competitive advantage over less active terminals in establishing synaptic connections. This principle is known to apply to fibers within a single neuronal population competing for a common target domain. Here we uncover an additional rule that applies when two neuronal populations compete for two contiguous territories. The cerebellar Purkinje cell dendrites have two different synaptic domains with spines innervated by two separate excitatory inputs, parallel fibers (PFs) and climbing fibers (CFs). Glutamate delta-2 receptors are normally present only on the PF spines where they are important for their innervation. After block of activity by tetrodotoxin, numerous new spines form in the CF domain and become innervated mainly by PFs; all spines, including those still innervated by the CFs, bear delta-2 receptors. Thus, in the absence of activity, PFs gain a competitive advantage over CFs. The entire dendritic arbor becomes a uniform territory with the molecular cues associated with the PFs. To access their proper territory and maintain synaptic contacts, CFs must be active and locally repress the cues of the competitor afferents.

Processing of shape defined by disparity in monkey inferior temporal cortex.

Neurons in the monkey inferior temporal cortex (IT) have been shown to respond to shapes defined by luminance, texture, or motion. In the present study, we determined whether IT neurons respond to shapes defined solely by binocular disparity, and if so, whether signals of disparity and other visual cues to define shape converge on single IT neurons. We recorded extracellular activity from IT neurons while monkeys performed a fixation task. Among the neurons that responded to at least one of eight random-dot stereograms (RDSs) containing different disparity-defined shapes, 21% varied their responses to different RDSs. Responses of most of the neurons were positively correlated between two sets of RDSs, which consisted of different dot patterns but defined the same set of eight shapes, whereas responses to RDSs and their monocular images were not correlated. This indicates that the response modulation for the eight RDSs reflects selectivity for shapes (or their component contours) defined by disparity, although responses were also affected by dot patterns per se. Among the neurons that showed selectivity for shapes defined by luminance or disparity, 44% were activated by both cues. Responses of these neurons to luminance-defined shapes and those to disparity-defined shapes were often positively correlated to each other. Furthermore the stimulus rank, which was determined by the magnitude of responses to shapes, generally matched between these cues. The same held true between disparity and texture cues. The results suggest that the signals of disparity, luminance, and texture cues to define the shapes converge on a population of single IT neurons to produce the selectivity for shapes.

Neurogenesis in the superior olivary complex in the rat

In a previous paper we provided evidence that crossed projection neurons are generated earlier than uncrossed projection neurons in the lateral superior olive. The aim of the present study was to determine if other major nuclei of the superior olivary complex (SOC), the medial superior olivary (MSO), the superior paraolivary (SPN) and the medial trapezoid (MTB) nuclei, are distinguished by their neuronal constitutions of birthdates. Pregnant rats were injected intraperitoneally with 5-bromodeoxyuridine (BrdU), the thymidine analogue, to label the neurons on one of the embryonic (E) days E11–E16. When the progeny rats reached adulthood, the brains were processed for BrdU immunohistochemistry. The MSO was mostly composed of neurons generated on E12 (95%). The remaining neurons in the MSO completed their neurogenesis by E13. The SPN neurons were generated from E12 to E14 with a peak on E13 (80%). Regardless of the morphological heterogeneity, the SPN consisted of a single population of neurons in terms of neurogenesis. The MTB neurons were generated from E13 to E16 with a peak on E14 (73%). In contrast to the previous assumption, no topographical relationship existed between neurogenesis and tonotopicity within each nucleus of the SOC.

Brain-Derived Neurotrophic Factor Differentially Regulates Retinal Ganglion Cell Dendritic and Axonal ArborizationIn Vivo

Expression of the neurotrophin brain-derived neurotrophic factor (BDNF) and its receptor trkB in the ganglion cell layer of the Xenopus retina during retinal ganglion cell (RGC) dendritic arborization indicates that BDNF is spatially and temporally available to influence RGC morphological differentiation (; ). BDNF promotes RGC axon arborization in vivo by acting as a target-derived trophic factor (). To determine whether BDNF also acts locally to regulate RGC dendritic development in vivo, we altered retinal neurotrophin levels at the onset of dendritic arborization and assessed the resulting arbor morphologies of RGCs retrogradely labeled with fluorescent dextrans. Injecting neurotrophins or BDNF function-blocking antibodies coupled to microspheres provided local alterations of retinal neurotrophin levels. BDNF significantly decreased RGC dendritic arbor complexity, whereas neutralizing endogenous BDNF levels with function-blocking antibodies significantly increased dendritic arbor complexity. RGCs exposed to other neurotrophins, as well as RGCs in retinae treated with BDNF but in areas not directly exposed to the neurotrophin, developed dendritic arbors that were indistinguishable from controls, indicating that exogenous BDNF acts specifically and locally. In the tectum, where RGC axons arborize, BDNF had opposite effects. BDNF significantly increased RGC axon arbor complexity and anti-BDNF reduced RGC arborization. Thus, BDNF reduces RGC dendritic arborization within the retina and increases axon arborization in the tectum. These results indicate that BDNF can differentially modulate axonal and dendritic arborization within a single neuronal population in opposing manners and raise the possibility that differential modulation by a neurotrophic factor finely tunes the morphological differentiation program of a neuron.

Parallel pathways mediating manual dexterity in the macaque.

Transmission of information along appropriately structured parallel pathways ensures that a great deal of information can be transferred from the source to the target very quickly, and with great security-essential features of any motor control system. Studies over the last two decades have established that the corticospinal and corticocerebellar pathways mediating manual dexterity in the primate are structurally organized to sustain the parallel transmission of sensorimotor information in multiple pathways. Serial, hierarchical control systems now seem insufficient to regulate voluntary hand movements. To achieve the required coordination, and precision and speed of execution, they must be combined with parallel control systems, which themselves incorporate elaborate feedforward and feedback controls. To illustrate these issues, two aspects of the structural organization of parallel sensorimotor pathways mediating manual dexterity in the macaque are reviewed. First, we examine the structure of the multiple corticospinal neuron subpopulations projecting from different areas of the frontoparietal cortex and how they are modified following hemisection of the cervical spinal cord. The remarkable recovery of hand function following spinal hemisection, despite the absence of any structural 'bridging' of the interrupted spinal pathways, and the fact that this is accountable in a parallel but not in a purely serial transmission system, are then reviewed. The second aspect of parallel distributed transmission examined is its occurrence within a single population of relay neurons. Our recent structural analysis of the somatic/dendritic organization of rubrospinal neurons in macaque red nucleus is used. The very large dendritic fields of individual neurons, extending over one-third or more of the nucleus, provide a framework for extracting precise somatotopic information from an input population whose axon terminal arbors overlap extensively, and, which, without effective filtering, would provide poor spatial resolution.