RESUMEN
Social interactions involve complex decision-making tasks that are shaped by dynamic, mutual feedback between participants. An open question is whether and how emergent properties may arise across brains of socially interacting individuals to influence social decisions. By simultaneously performing microendoscopic calcium imaging in pairs of socially interacting mice, we find that animals exhibit interbrain correlations of neural activity in the prefrontal cortex that are dependent on ongoing social interaction. Activity synchrony arises from two neuronal populations that separately encode one's own behaviors and those of the social partner. Strikingly, interbrain correlations predict future social interactions as well as dominance relationships in a competitive context. Together, our study provides conclusive evidence for interbrain synchrony in rodents, uncovers how synchronization arises from activity at the single-cell level, and presents a role for interbrain neural activity coupling as a property of multi-animal systems in coordinating and sustaining social interactions between individuals.
Asunto(s)
Encéfalo/metabolismo , Neuronas/metabolismo , Animales , Señalización del Calcio , Conducta Competitiva/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Corteza Prefrontal/metabolismo , Análisis de Componente Principal , Predominio SocialRESUMEN
Working memory, the process through which information is transiently maintained and manipulated over a brief period, is essential for most cognitive functions1-4. However, the mechanisms underlying the generation and evolution of working-memory neuronal representations at the population level over long timescales remain unclear. Here, to identify these mechanisms, we trained head-fixed mice to perform an olfactory delayed-association task in which the mice made decisions depending on the sequential identity of two odours separated by a 5 s delay. Optogenetic inhibition of secondary motor neurons during the late-delay and choice epochs strongly impaired the task performance of the mice. Mesoscopic calcium imaging of large neuronal populations of the secondary motor cortex (M2), retrosplenial cortex (RSA) and primary motor cortex (M1) showed that many late-delay-epoch-selective neurons emerged in M2 as the mice learned the task. Working-memory late-delay decoding accuracy substantially improved in the M2, but not in the M1 or RSA, as the mice became experts. During the early expert phase, working-memory representations during the late-delay epoch drifted across days, while the stimulus and choice representations stabilized. In contrast to single-plane layer 2/3 (L2/3) imaging, simultaneous volumetric calcium imaging of up to 73,307 M2 neurons, which included superficial L5 neurons, also revealed stabilization of late-delay working-memory representations with continued practice. Thus, delay- and choice-related activities that are essential for working-memory performance drift during learning and stabilize only after several days of expert performance.
Asunto(s)
Consolidación de la Memoria , Memoria a Corto Plazo , Práctica Psicológica , Animales , Femenino , Masculino , Ratones , Calcio/metabolismo , Conducta de Elección/fisiología , Consolidación de la Memoria/fisiología , Memoria a Corto Plazo/fisiología , Ratones Endogámicos C57BL , Corteza Motora/fisiología , Corteza Motora/citología , Neuronas Motoras/fisiología , Odorantes/análisis , Optogenética , Desempeño Psicomotor/fisiología , Olfato/fisiología , Factores de TiempoRESUMEN
Genetically encoded voltage indicators (GEVIs) enable optical recording of electrical signals in the brain, providing subthreshold sensitivity and temporal resolution not possible with calcium indicators. However, one- and two-photon voltage imaging over prolonged periods with the same GEVI has not yet been demonstrated. Here, we report engineering of ASAP family GEVIs to enhance photostability by inversion of the fluorescence-voltage relationship. Two of the resulting GEVIs, ASAP4b and ASAP4e, respond to 100-mV depolarizations with ≥180% fluorescence increases, compared with the 50% fluorescence decrease of the parental ASAP3. With standard microscopy equipment, ASAP4e enables single-trial detection of spikes in mice over the course of minutes. Unlike GEVIs previously used for one-photon voltage recordings, ASAP4b and ASAP4e also perform well under two-photon illumination. By imaging voltage and calcium simultaneously, we show that ASAP4b and ASAP4e can identify place cells and detect voltage spikes with better temporal resolution than commonly used calcium indicators. Thus, ASAP4b and ASAP4e extend the capabilities of voltage imaging to standard one- and two-photon microscopes while improving the duration of voltage recordings.
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Encéfalo , Calcio , Animales , Ratones , Iluminación , Microscopía , FotonesRESUMEN
Although many genes predisposing to autism spectrum disorders (ASD) have been identified, the biological mechanism(s) remain unclear. Mouse models based on human disease-causing mutations provide the potential for understanding gene function and novel treatment development. Here, we characterize a mouse knockout of the Cntnap2 gene, which is strongly associated with ASD and allied neurodevelopmental disorders. Cntnap2(-/-) mice show deficits in the three core ASD behavioral domains, as well as hyperactivity and epileptic seizures, as have been reported in humans with CNTNAP2 mutations. Neuropathological and physiological analyses of these mice before the onset of seizures reveal neuronal migration abnormalities, reduced number of interneurons, and abnormal neuronal network activity. In addition, treatment with the FDA-approved drug risperidone ameliorates the targeted repetitive behaviors in the mutant mice. These data demonstrate a functional role for CNTNAP2 in brain development and provide a new tool for mechanistic and therapeutic research in ASD.
Asunto(s)
Trastorno Autístico/genética , Encéfalo/crecimiento & desarrollo , Modelos Animales de Enfermedad , Proteínas de la Membrana/metabolismo , Ratones , Proteínas del Tejido Nervioso/metabolismo , Animales , Trastorno Autístico/patología , Encéfalo/metabolismo , Encéfalo/patología , Movimiento Celular , Epilepsia/genética , Humanos , Interneuronas/metabolismo , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Ratones Noqueados , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/genética , Neuronas/patologíaRESUMEN
The ventrolateral preoptic area (VLPO) contains GABAergic sleep-active neurons. However, the extent to which these neurons are involved in expressing spontaneous sleep and homeostatic sleep regulatory demands is not fully understood. We used calcium (Ca2+) imaging to characterize the activity dynamics of VLPO neurons, especially those expressing the vesicular GABA transporter (VGAT) across spontaneous sleep-waking and in response to homeostatic sleep demands. The VLPOs of wild-type and VGAT-Cre mice were transfected with GCaMP6, and the Ca2+ fluorescence of unidentified (UNID) and VGAT cells was recorded during spontaneous sleep-waking and 3 h of sleep deprivation (SD) followed by 1 h of recovery sleep. Although both VGAT and UNID neurons exhibited heterogeneous Ca2+ fluorescence across sleep-waking, the majority of VLPO neurons displayed increased activity during nonREM/REM (VGAT, 120/303; UNID, 39/106) and REM sleep (VGAT, 32/303; UNID, 19/106). Compared to the baseline waking, VLPO sleep-active neurons (n = 91) exhibited higher activity with increasing SD that remained elevated during the recovery period. These neurons also exhibited increased Ca2+ fluorescence during nonREM sleep, marked by increased slow-wave activity and REM sleep during recovery after SD. These findings support the notion that VLPO sleep-active neurons, including GABAergic neurons, are components of neuronal circuitry that mediate spontaneous sleep and homeostatic responses to sustained wakefulness.
Asunto(s)
Calcio , Sueño , Ratones , Animales , Sueño/fisiología , Neuronas GABAérgicas/fisiología , Privación de Sueño , Sueño REM , Área Preóptica , Calcio de la DietaRESUMEN
Recent studies suggest that a shared neural ensemble may link distinct memories encoded close in time. According to the memory allocation hypothesis, learning triggers a temporary increase in neuronal excitability that biases the representation of a subsequent memory to the neuronal ensemble encoding the first memory, such that recall of one memory increases the likelihood of recalling the other memory. Here we show in mice that the overlap between the hippocampal CA1 ensembles activated by two distinct contexts acquired within a day is higher than when they are separated by a week. Several findings indicate that this overlap of neuronal ensembles links two contextual memories. First, fear paired with one context is transferred to a neutral context when the two contexts are acquired within a day but not across a week. Second, the first memory strengthens the second memory within a day but not across a week. Older mice, known to have lower CA1 excitability, do not show the overlap between ensembles, the transfer of fear between contexts, or the strengthening of the second memory. Finally, in aged mice, increasing cellular excitability and activating a common ensemble of CA1 neurons during two distinct context exposures rescued the deficit in linking memories. Taken together, these findings demonstrate that contextual memories encoded close in time are linked by directing storage into overlapping ensembles. Alteration of these processes by ageing could affect the temporal structure of memories, thus impairing efficient recall of related information.
Asunto(s)
Región CA1 Hipocampal/citología , Región CA1 Hipocampal/fisiología , Memoria/fisiología , Neuronas/fisiología , Animales , Calcio/análisis , Miedo , Masculino , Recuerdo Mental/fisiología , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Factores de TiempoRESUMEN
Thus far, optical recording of neuronal activity in freely behaving animals has been limited to a thin axial range. We present a head-mounted miniaturized light-field microscope (MiniLFM) capable of capturing neuronal network activity within a volume of 700 × 600 × 360 µm3 at 16 Hz in the hippocampus of freely moving mice. We demonstrate that neurons separated by as little as ~15 µm and at depths up to 360 µm can be discriminated.
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Hipocampo/citología , Hipocampo/fisiología , Miniaturización/instrumentación , Neuronas/fisiología , Animales , Microscopía Intravital/instrumentación , Microscopía Intravital/métodos , Ratones , Imagen Óptica/instrumentación , Imagen Óptica/métodosRESUMEN
In the version of this Brief Communication originally published online, ref. 21 included details for a conference paper (Pegard, N. C. et al. Paper presented at Novel Techniques in Microscopy: Optics in the Life Sciences, Vancouver, BC, Canada, 12-15 April 2015). The correct reference is the following: Pégard, N. C. et al. Optica 3, 517-524 (2016). This error has been corrected in the print, HTML and PDF versions of the paper.
RESUMEN
Huntington's disease (HD) is a neurodegenerative disorder characterized by involuntary movements, cognitive deficits, and psychiatric disturbances. Although evidence indicates that projections from motor cortical areas play a key role in the development of dysfunctional striatal activity and motor phenotype, little is known about the changes in cortical microcircuits and their role in the development of the HD phenotype. Here we used two-photon laser-scanning microscopy to evaluate network dynamics of motor cortical neurons in layers II/III in behaving transgenic R6/2 and knock-in Q175+/- mice. Symptomatic R6/2 mice displayed increased motion manifested by a significantly greater number of motion epochs, whereas symptomatic Q175 mice displayed decreased motion. In both models, calcium transients in symptomatic mice displayed reduced amplitude, suggesting decreased bursting activity. Changes in frequency were genotype- and time-dependent; for R6/2 mice, the frequency was reduced during both motion and nonmotion, whereas in symptomatic Q175 mice, the reduction only occurred during nonmotion. In presymptomatic Q175 mice, frequency was increased during both behavioral states. Interneuronal correlation coefficients were generally decreased in both models, suggesting disrupted interneuronal communication in HD cerebral cortex. These results indicate similar and contrasting effects of the HD mutation on cortical ensemble activity depending on mouse model and disease stage.
Asunto(s)
Calcio , Modelos Animales de Enfermedad , Enfermedad de Huntington/diagnóstico por imagen , Enfermedad de Huntington/genética , Corteza Motora/diagnóstico por imagen , Red Nerviosa/diagnóstico por imagen , Animales , Calcio/metabolismo , Femenino , Enfermedad de Huntington/metabolismo , Masculino , Ratones , Ratones Transgénicos , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Corteza Motora/metabolismo , Neuronas Motoras/metabolismo , Red Nerviosa/metabolismoRESUMEN
Despite substantial recent progress in network neuroscience, the impact of stroke on the distinct features of reorganizing neuronal networks during recovery has not been defined. Using a functional connections-based approach through 2-photon in vivo calcium imaging at the level of single neurons, we demonstrate for the first time the functional connectivity maps during motion and nonmotion states, connection length distribution in functional connectome maps and a pattern of high clustering in motor and premotor cortical networks that is disturbed in stroke and reconstitutes partially in recovery. Stroke disrupts the network topology of connected inhibitory and excitatory neurons with distinct patterns in these 2 cell types and in different cortical areas. These data indicate that premotor cortex displays a distinguished neuron-specific recovery profile after stroke.
Asunto(s)
Actividad Motora , Corteza Motora/fisiopatología , Neuronas/fisiología , Recuperación de la Función , Accidente Cerebrovascular/fisiopatología , Animales , Señalización del Calcio , Masculino , Ratones Transgénicos , Imagen ÓpticaRESUMEN
Although whole-organism calcium imaging in small and semi-transparent animals has been demonstrated, capturing the functional dynamics of large-scale neuronal circuits in awake behaving mammals at high speed and resolution has remained one of the main frontiers in systems neuroscience. Here we present a method based on light sculpting that enables unbiased single- and dual-plane high-speed (up to 160 Hz) calcium imaging as well as in vivo volumetric calcium imaging of a mouse cortical column (0.5 mm × 0.5 mm × 0.5 mm) at single-cell resolution and fast volume rates (3-6 Hz). We achieved this by tailoring the point-spread function of our microscope to the structures of interest while maximizing the signal-to-noise ratio using a home-built fiber laser amplifier with pulses that are synchronized to the imaging voxel speed. This enabled in vivo recording of calcium dynamics of several thousand neurons across cortical layers and in the hippocampus of awake behaving mice.
Asunto(s)
Señalización del Calcio/fisiología , Calcio/metabolismo , Corteza Cerebral/metabolismo , Hipocampo/metabolismo , Imagen Molecular/métodos , Neuronas/metabolismo , Animales , Conducta Animal/fisiología , Ratones , Microscopía Confocal , Fotones , Factores de TiempoRESUMEN
Neuroimaging techniques applied to a variety of organisms-from zebrafish, to rodents to humans-can offer valuable insights into neuronal network properties and their dysfunction in epilepsy. A wide range of imaging methods used to monitor neuronal circuits and networks during evoked seizures in animal models and advances in functional magnetic resonance imaging (fMRI) applied to patients with epilepsy were discussed during the XIV Workshop on Neurobiology of Epilepsy (XIV WONOEP) organized in 2017 by the Neurobiology Commission of the International League Against Epilepsy (ILAE). We review the growing number of technological approaches developed, as well as the current state of knowledge gained from studies applying these advanced imaging approaches to epilepsy research.
Asunto(s)
Encéfalo/diagnóstico por imagen , Epilepsia/diagnóstico por imagen , Neuroimagen Funcional , Animales , Epilepsia Refractaria/diagnóstico por imagen , Educación , Epilepsia del Lóbulo Temporal/diagnóstico por imagen , Neuroimagen Funcional/métodos , HumanosRESUMEN
Low-frequency membrane potential (Vm) oscillations were once thought to only occur in sleeping and anesthetized states. Recently, low-frequency Vm oscillations have been described in inactive awake animals, but it is unclear whether they shape sensory processing in neurons and whether they occur during active awake behavioral states. To answer these questions, we performed two-photon guided whole-cell Vm recordings from primary visual cortex layer 2/3 excitatory and inhibitory neurons in awake mice during passive visual stimulation and performance of visual and auditory discrimination tasks. We recorded stereotyped 3-5 Hz Vm oscillations where the Vm baseline hyperpolarized as the Vm underwent high amplitude rhythmic fluctuations lasting 1-2 s in duration. When 3-5 Hz Vm oscillations coincided with visual cues, excitatory neuron responses to preferred cues were significantly reduced. Despite this disruption to sensory processing, visual cues were critical for evoking 3-5 Hz Vm oscillations when animals performed discrimination tasks and passively viewed drifting grating stimuli. Using pupillometry and animal locomotive speed as indicators of arousal, we found that 3-5 Hz oscillations were not restricted to unaroused states and that they occurred equally in aroused and unaroused states. Therefore, low-frequency Vm oscillations play a role in shaping sensory processing in visual cortical neurons, even during active wakefulness and decision making.SIGNIFICANCE STATEMENT A neuron's membrane potential (Vm) strongly shapes how information is processed in sensory cortices of awake animals. Yet, very little is known about how low-frequency Vm oscillations influence sensory processing and whether they occur in aroused awake animals. By performing two-photon guided whole-cell recordings from layer 2/3 excitatory and inhibitory neurons in the visual cortex of awake behaving animals, we found visually evoked stereotyped 3-5 Hz Vm oscillations that disrupt excitatory responsiveness to visual stimuli. Moreover, these oscillations occurred when animals were in high and low arousal states as measured by animal speed and pupillometry. These findings show, for the first time, that low-frequency Vm oscillations can significantly modulate sensory signal processing, even in awake active animals.
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Relojes Biológicos/fisiología , Potenciales Evocados Visuales/fisiología , Potenciales de la Membrana/fisiología , Neuronas/fisiología , Corteza Visual/fisiología , Percepción Visual/fisiología , Vigilia/fisiología , Animales , Conducta Animal/fisiología , Ondas Encefálicas/fisiología , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Análisis y Desempeño de TareasRESUMEN
UNLABELLED: Arginase 1 deficiency is a urea cycle disorder associated with hyperargininemia, spastic diplegia, loss of ambulation, intellectual disability, and seizures. To gain insight on how loss of arginase expression affects the excitability and synaptic connectivity of the cortical neurons in the developing brain, we used anatomical, ultrastructural, and electrophysiological techniques to determine how single-copy and double-copy arginase deletion affects cortical circuits in mice. We find that the loss of arginase 1 expression results in decreased dendritic complexity, decreased excitatory and inhibitory synapse numbers, decreased intrinsic excitability, and altered synaptic transmission in layer 5 motor cortical neurons. Hepatic arginase 1 gene therapy using adeno-associated virus rescued nearly all these abnormalities when administered to neonatal homozygous knock-out animals. Therefore, gene therapeutic strategies can reverse physiological and anatomical markers of arginase 1 deficiency and therefore may be of therapeutic benefit for the neurological disabilities in this syndrome. SIGNIFICANCE STATEMENT: These studies are one of the few investigations to try to understand the underlying neurological dysfunction that occurs in urea cycle disorders and the only to examine arginase deficiency. We have demonstrated by multiple modalities that, in murine layer 5 cortical neurons, a gradation of abnormalities exists based on the functional copy number of arginase: intrinsic excitability is altered, there is decreased density in asymmetrical and perisomatic synapses, and analysis of the dendritic complexity is lowest in the homozygous knock-out. With neonatal administration of adeno-associated virus expressing arginase, there is near-total recovery of the abnormalities in neurons and cortical circuits, supporting the concept that neonatal gene therapy may prevent the functional abnormalities that occur in arginase deficiency.
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Arginasa/uso terapéutico , Terapia Genética , Hiperargininemia/patología , Hiperargininemia/terapia , Corteza Motora/fisiología , Recuperación de la Función/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Amoníaco/sangre , Animales , Animales Recién Nacidos , Arginasa/genética , Arginasa/metabolismo , Modelos Animales de Enfermedad , Hiperargininemia/sangre , Técnicas In Vitro , Ratones , Ratones Transgénicos , Corteza Motora/citología , Corteza Motora/ultraestructura , Red Nerviosa/patología , Red Nerviosa/fisiología , Red Nerviosa/ultraestructura , Neuronas/fisiología , Neuronas/ultraestructura , Picrotoxina/farmacología , Bloqueadores de los Canales de Sodio/farmacología , Sinapsis/ultraestructura , Tetrodotoxina/farmacologíaAsunto(s)
Microscopía/instrumentación , Miniaturización , Neuroimagen/instrumentación , Animales , Cabeza , RatonesRESUMEN
As the major input to the basal ganglia, the striatum is innervated by a wide range of other areas. Overlapping input from these regions is speculated to influence temporal correlations among striatal ensembles. However, the network dynamics among behaviorally related neural populations in the striatum has not been extensively studied. We used large-scale neural recordings to monitor activity from striatal ensembles in mice undergoing Pavlovian reward conditioning. A subpopulation of putative medium spiny projection neurons (MSNs) was found to discriminate between cues that predicted the delivery of a reward and cues that predicted no specific outcome. These cells were preferentially located in lateral subregions of the striatum. Discriminating MSNs were more spontaneously active and more correlated than their nondiscriminating counterparts. Furthermore, discriminating fast spiking interneurons (FSIs) represented a highly prevalent group in the recordings, which formed a strongly correlated network with discriminating MSNs. Spike time cross-correlation analysis showed the existence of synchronized activity among FSIs and feedforward inhibitory modulation of MSN spiking by FSIs. These findings suggest that populations of functionally specialized (cue-discriminating) striatal neurons have distinct network dynamics that sets them apart from nondiscriminating cells, potentially to facilitate accurate behavioral responding during associative reward learning.
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Condicionamiento Clásico , Cuerpo Estriado/fisiología , Discriminación en Psicología , Neuronas/fisiología , Recompensa , Animales , Cuerpo Estriado/citología , Señales (Psicología) , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
De novo phosphatase and tensin homolog on chromosome ten (PTEN) mutations are a cause of sporadic autism. How single-copy loss of PTEN alters neural function is not understood. Here we report that Pten haploinsufficiency increases the expression of small-conductance calcium-activated potassium channels. The resultant augmentation of this conductance increases the amplitude of the afterspike hyperpolarization, causing a decrease in intrinsic excitability. In vivo, this change in intrinsic excitability reduces evoked firing rates of cortical pyramidal neurons but does not alter receptive field tuning. The decreased in vivo firing rate is not associated with deficits in the dendritic integration of synaptic input or with changes in dendritic complexity. These findings identify calcium-activated potassium channelopathy as a cause of cortical dysfunction in the PTEN model of autism and provide potential molecular therapeutic targets.
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Trastorno Autístico/genética , Canalopatías/fisiopatología , Fosfohidrolasa PTEN/genética , Canales de Potasio de Pequeña Conductancia Activados por el Calcio/metabolismo , Análisis de Varianza , Animales , Trastorno Autístico/fisiopatología , Western Blotting , Canalopatías/genética , Hemicigoto , Humanos , Ratones , Mutación/genética , Técnicas de Placa-Clamp , Tractos Piramidales/citología , Tractos Piramidales/fisiología , Canales de Potasio de Pequeña Conductancia Activados por el Calcio/genéticaRESUMEN
PURPOSE OF REVIEW: This review discusses the ways that rodent models of autism spectrum disorders (ASDs) have been used to gain critical information about convergent molecular pathways, the mechanisms underlying altered microcircuit structure and function, and as a screen for potential cutting edge-treatments for ASDs. RECENT FINDINGS: There is convergent evidence that impaired developmental pruning of connections may be a common finding among several mouse models of ASDs. Recent studies have uncovered impaired autophagy by pathological mTOR activation as a potential contributor to microcircuit dysfunction and behavior. ASD-related disinhibition and exaggerated synaptic plasticity in multiple distinct circuits in cortex and reward circuits in striatum also contribute to social dysfunction and repetitive behaviors. New exciting molecular therapeutic techniques have reversed cognitive deficits in models of ASD, indicating that mouse models could be used for preclinical translational studies of new treatments. SUMMARY: Rodent models of ASDs coupled to new emerging technologies for genome editing, cell-specific functional and structural imaging, and neuronal activity manipulation will yield critical insights into ASD pathogenesis and fuel the emergence of new treatments.
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Trastorno del Espectro Autista/genética , Modelos Animales de Enfermedad , Predisposición Genética a la Enfermedad , Plasticidad Neuronal/genética , Neuronas/citología , Animales , Humanos , Serina-Treonina Quinasas TOR/genéticaRESUMEN
Endogenous brain rhythms occurring at various frequencies and associated with distinct behavioral states provide multiscale temporal windows that enable cells to time their spiking activity with high precision, which is thought to be important for the coding of information in neuronal circuits. However, although the selective timing of GABAergic inputs to specific spatial domains of principal cells are known to play key roles in network oscillations, the in vivo firing patterns of distinct hippocampal interneurons in awake animals are not known. Here we used a combination of juxtacellular labeling techniques with recordings from anesthesia-free, head-fixed mice running or resting on a spherical treadmill to study the oscillation-dependent discharges by two major interneuronal subtypes, the perisomatically projecting parvalbumin-positive basket cells (PVBCs) and distal dendritically projecting oriens lacunosum moleculare (OLM) cells. Recordings of the spiking activity of post hoc-identified CA1 interneurons during theta (5-10 Hz), gamma (25-90Hz), epsilon ("high-gamma"; 90-130 Hz), and ripple (130-200 Hz) oscillations revealed both cell type- and behavioral state-dependent entrainments of PVBC and OLM cell discharges in awake mice. Our results in awake mice differed in several respects from previous data on interneuronal discharge patterns in anesthetized animals. In addition, our results demonstrate a form of frequency-invariant, cell type-specific temporal ordering of inhibitory inputs in which PVBC-derived perisomatic inhibition is followed by OLM cell-generated distal dendritic inhibition during each of the network oscillation bands studied, spanning more than an order of magnitude in frequencies.
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Potenciales de Acción/fisiología , Hipocampo/fisiología , Interneuronas/fisiología , Ritmo Teta/fisiología , Vigilia/fisiología , Animales , Mapeo Encefálico , Inmunohistoquímica , Interneuronas/ultraestructura , Ratones , Microelectrodos , Microscopía ElectrónicaRESUMEN
In vivo two-photon calcium imaging would benefit from the use of multiple excitation beams to increase scanning speed, signal-to-noise ratio and field of view or to image different axial planes simultaneously. Using spatiotemporal multiplexing we circumvented light-scattering ambiguity inherent to deep-tissue multifocal two-photon microscopy. We demonstrate calcium imaging at multiple axial planes in the intact mouse brain to monitor network activity of ensembles of cortical neurons in three spatial dimensions.