Emergence of dominant initiation sites for interictal spikes in rat neocortex.

Neuronal populations with unbalanced inhibition can generate interictal spikes (ISs), where each IS starts from a small initiation site and then spreads activation across a larger area. We used in vivo voltage-sensitive dye imaging to map the initiation site of ISs in rat visual cortex disinhibited by epidural application of bicuculline methiodide. Immediately after the application of bicuculline, the IS initiation sites were widely distributed over the entire disinhibited area. After ∼ 10 min, a small number of sites became "dominant" and initiated the majority of the ISs throughout the course of imaging. Such domination also occurred in cortical slices, which lack long-range connections between the cortex and subcortical structures. This domination of IS initiation sites may allow timing-related plasticity mechanisms to provide a spatial organization where connections projecting outward from the dominant initiation site become strengthened. Understanding the spatiotemporal organization of IS initiation sites may contribute to our understanding of epileptogenesis in its very early stages, because a dominant IS initiation site with strengthened outward connectivity may ultimately develop into a seizure focus.

Wave propagation of cortical population activity under urethane anesthesia is state dependent.

BACKGROUND: Propagating waves of excitation have been observed extensively in the neocortex, during both spontaneous and sensory-evoked activity, and they play a critical role in spatially organizing information processing. However, the state-dependence of these spatiotemporal propagation patterns is largely unexplored. In this report, we use voltage-sensitive dye imaging in the rat visual cortex to study the propagation of spontaneous population activity in two discrete cortical states induced by urethane anesthesia. RESULTS: While laminar current source density patterns of spontaneous population events in these two states indicate a considerable degree of similarity in laminar networks, lateral propagation in the more active desynchronized state is approximately 20% faster than in the slower synchronized state. Furthermore, trajectories of wave propagation exhibit a strong anisotropy, but the preferred direction is different depending on cortical state. CONCLUSIONS: Our results show that horizontal wave propagation of spontaneous neural activity is largely dependent on the global activity states of local cortical circuits.

Compression and reflection of visually evoked cortical waves.

Neuronal interactions between primary and secondary visual cortical areas are important for visual processing, but the spatiotemporal patterns of the interaction are not well understood. We used voltage-sensitive dye imaging to visualize neuronal activity in rat visual cortex and found visually evoked waves propagating from V1 to other visual areas. A primary wave originated in the monocular area of V1 and was "compressed" when propagating to V2. A reflected wave initiated after compression and propagated backward into V1. The compression occurred at the V1/V2 border, and local GABAA inhibition is important for the compression. The compression/reflection pattern provides a two-phase modulation: V1 is first depolarized by the primary wave, and then V1 and V2 are simultaneously depolarized by the reflected and primary waves, respectively. The compression/reflection pattern only occurred for evoked waves and not for spontaneous waves, suggesting that it is organized by an internal mechanism associated with visual processing.

An optically transparent multi-electrode array for combined electrophysiology and optophysiology at the mesoscopic scale.

OBJECTIVE: A number of tissue penetrating opto-electrodes to simultaneously record and optogenetically influence brain activity have been developed. For experiments at the surface of the brain, such as electrocorticogram (ECoG) recordings and surface optogenetics, fewer devices have been described and no device has found widespread adoption for neuroscientific experiments. One issue slowing adoption is the complexity and fragility of existing devices, typically based on transparent electrode materials like graphene and indium-tin oxide (ITO). We focused here on improving existing processes based on metal traces and polyimide (PI), which produce more robust and cost-effective devices, to develop a multi-electrode array for optophysiology. APPROACH: The most widely used substrate material for surface electrodes, PI, has seen little use for optophysiologicalµECoG/ECoG arrays. This is due to its lack of transparency at optogenetically relevant short wavelengths. Here we use very thin layers of PI in combination with chrome-gold-platinum electrodes to achieve the necessary substrate transparency and high mechanical flexibility in a device that still rejects light artifacts well. MAIN RESULTS: The manufactured surface arrays have a thickness of only 6.5 µm, resulting in 80% transparency for blue light. We demonstrate immunity against opto-electric artifacts, long term stability and biocompatibility as well as suitability for optical voltage imaging. The biocompatible arrays are capable of recording stable ECoGs over months without any measurable degradation and can be used to map the tonotopic organization of the curved rodent auditory cortex. SIGNIFICANCE: Our novel probes combine proven materials and processing steps to create optically near-transparent electrode arrays with superior longevity. In contrast to previous opto-electrodes, our probes are simple to manufacture, robust, offer long-term stability, and are a practical engineering solution for optophysiological experiments not requiring transparency of the electrode sites themselves.

Author Correction: Increasing neurogenesis refines hippocampal activity rejuvenating navigational learning strategies and contextual memory throughout life.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Increasing neurogenesis refines hippocampal activity rejuvenating navigational learning strategies and contextual memory throughout life.

Functional plasticity of the brain decreases during ageing causing marked deficits in contextual learning, allocentric navigation and episodic memory. Adult neurogenesis is a prime example of hippocampal plasticity promoting the contextualisation of information and dramatically decreases during ageing. We found that a genetically-driven expansion of neural stem cells by overexpression of the cell cycle regulators Cdk4/cyclinD1 compensated the age-related decline in neurogenesis. This triggered an overall inhibitory effect on the trisynaptic hippocampal circuit resulting in a changed profile of CA1 sharp-wave ripples known to underlie memory consolidation. Most importantly, increased neurogenesis rescued the age-related switch from hippocampal to striatal learning strategies by rescuing allocentric navigation and contextual memory. Our study demonstrates that critical aspects of hippocampal function can be reversed in old age, or compensated throughout life, by exploiting the brain's endogenous reserve of neural stem cells.

Crossmodal propagation of sensory-evoked and spontaneous activity in the rat neocortex.

In the cortex, neural responses to crossmodal stimulation are seen both in higher association areas and in primary sensory areas, and are thought to play a role in integration of crossmodal sensations. We used voltage-sensitive dye imaging (VSDI) to study the spatiotemporal characteristics of such crossmodal neural activity. We imaged three cortical regions in rat: primary visual cortex (V1), barrel field of primary somatosensory cortex (S1bf) and parietal association area (PA, flanked by V1 and S1bf). We find that sensory-evoked population activity can propagate in the form of a distinct propagating wave, robustly in either crossmodal direction. In single trials, the waveforms changed continuously during propagation, with dynamic variability from trial to trial, which we interpret as evidence for cortical involvement in the spreading process. To further characterize the functional anatomy of PA, we also studied the propagation of spontaneous sleep-like waves in this area. Using a novel flow-detection algorithm, we detected a propagation bias within PA of spontaneous waves--these tend to propagate parallel to the crossmodal axis, rather than orthogonal to it. Taken together, these findings demonstrate that intracortical networks show pre-attentive crossmodal propagation of activity, and suggest a potential mechanism for the establishment of crossmodal integration.

Measurement, modeling, and prediction of temperature rise due to optogenetic brain stimulation.

Optogenetics is one of the most important techniques in neurophysiology, with potential clinical applications. However, the strong light needed may cause harmful temperature rises. So far, there are no methods to reliably estimate brain heating and safe limits in actual optogenetic experiments. We used thermal imaging to directly measure such temperature rises at the surface of live mouse brains during laser illumination with wavelengths and intensities typical for optogenetics. We then modeled the temperature rise with a simple logarithmic model. Our results indicate that previous finite-element models can underestimate temperature increases by an order of magnitude. We validate our empirical model by predicting the temperature rise caused by pulsed stimulation paradigms. These predictions fit closely to the empirical data and constitute a better estimate of real temperature increases. Additionally, we provide a web-based app for easy calculation that can be used as a tool for safe design of optogenetic experiments.

Periodic Lateralized Epileptiform Discharges (PLEDs) in Patients With Neurosyphilis and HIV Infection.

Periodic lateralized epileptiform discharges (PLEDs) are an electroencephalographic pattern recorded in the setting of a variety of brain abnormalities. It is best recognized for its association with acute viral encephalitis, stroke, tumor, or latestatus epilepticus. However, there are other conditions that have been recognized as the underlying pathology for PLEDs such as alcohol withdrawal, Creutzfeldt-Jacob disease, anoxic brain injury, and hemiplegic migraine. However, there are only rare case reports of PLEDs in patients with neurosyphilis. Here, we report 2 patients presenting with encephalopathy and seizures with PLEDs, ipsilateral or contralateral to their main brain magnetic resonance imaging abnormalities. Further workup revealed neurosyphilis in both patients, one in association with human immunodeficiency virus (HIV) infection. Given the increasing incidence of neurosyphilis with or without HIV infection, these cases suggest neurosyphilis as a consideration in the differential for patients presenting with PLEDs.

Development of the posterior basic rhythm in children with autism.

OBJECTIVE: Early detection of autism is critical for effective intervention, but currently, no simple screening tests are available. Furthermore, little is known about the development of brain dynamics in young children. We examine the early neurophysiological manifestations of autism by retrospectively analyzing EEG. In particular, we focus on maturation of the posterior basic rhythm (PBR), which is one of the most characteristic features of the normal EEG, and comprises a discrete functional state. METHODS: Subjects with a diagnosis of autism (n=74), as well as normal (n=134) and epileptic (n=108) controls, were extracted retrospectively from our digital EEG database. Segments with clear PBR were extracted, and standard signal analysis methods were used to calculate peak PBR frequency, power, and coherence. RESULTS: In our cohort, a subset of autistic children show accelerated development of the PBR, with early maturation especially in the 2- to 4-year old range. The overall coherence of PBR-specific activity is also lower in autistic children in our cohort. CONCLUSIONS: These findings provide evidence that autism is associated with accelerated development of the PBR. SIGNIFICANCE: These findings generate a clinical hypothesis for future prospective studies on the efficacy of these simple measures as a diagnostic or screening tool.

Asymmetric multisensory interactions of visual and somatosensory responses in a region of the rat parietal cortex.

Perception greatly benefits from integrating multiple sensory cues into a unified percept. To study the neural mechanisms of sensory integration, model systems are required that allow the simultaneous assessment of activity and the use of techniques to affect individual neural processes in behaving animals. While rodents qualify for these requirements, little is known about multisensory integration and areas involved for this purpose in the rodent. Using optical imaging combined with laminar electrophysiological recordings, the rat parietal cortex was identified as an area where visual and somatosensory inputs converge and interact. Our results reveal similar response patterns to visual and somatosensory stimuli at the level of current source density (CSD) responses and multi-unit responses within a strip in parietal cortex. Surprisingly, a selective asymmetry was observed in multisensory interactions: when the somatosensory response preceded the visual response, supra-linear summation of CSD was observed, but the reverse stimulus order resulted in sub-linear effects in the CSD. This asymmetry was not present in multi-unit activity however, which showed consistently sub-linear interactions. These interactions were restricted to a specific temporal window, and pharmacological tests revealed significant local intra-cortical contributions to this phenomenon. Our results highlight the rodent parietal cortex as a system to model the neural underpinnings of multisensory processing in behaving animals and at the cellular level.

Flow detection of propagating waves with temporospatial correlation of activity.

Voltage-sensitive dye imaging (VSDI) allows population patterns of cortical activity to be recorded with high temporal resolution, and recent findings ascribe potential significance to these spatial propagation patterns--both for normal cortical processing and in pathologies such as epilepsy. However, analysis of these spatiotemporal patterns has been mostly qualitative to date. In this report, we describe an algorithm to quantify fast local flow patterns of cortical population activation, as measured with VSDI. The algorithm uses correlation of temporal features across space, and therefore differs from conventional optical flow algorithms which use correlation of spatial features over time. This alternative approach allows us to take advantage of the characteristics of fast optical imaging data, which have very high temporal resolution but less spatial resolution. We verify the method both on artificial and biological data, and demonstrate its use.

Spiral wave dynamics in neocortex.

Although spiral waves are ubiquitous features of nature and have been observed in many biological systems, their existence and potential function in mammalian cerebral cortex remain uncertain. Using voltage-sensitive dye imaging, we found that spiral waves occur frequently in the neocortex in vivo, both during pharmacologically induced oscillations and during sleep-like states. While their life span is limited, spiral waves can modify ongoing cortical activity by influencing oscillation frequencies and spatial coherence and by reducing amplitude in the area surrounding the spiral phase singularity. During sleep-like states, the rate of occurrence of spiral waves varies greatly depending on brain states. These results support the hypothesis that spiral waves, as an emergent activity pattern, can organize and modulate cortical population activity on the mesoscopic scale and may contribute to both normal cortical processing and to pathological patterns of activity such as those found in epilepsy.

Normalization of voltage-sensitive dye signal with functional activity measures.

In general, signal amplitude in optical imaging is normalized using the well-established DeltaF/F method, where functional activity is divided by the total fluorescent light flux. This measure is used both directly, as a measure of population activity, and indirectly, to quantify spatial and spatiotemporal activity patterns. Despite its ubiquitous use, the stability and accuracy of this measure has not been validated for voltage-sensitive dye imaging of mammalian neocortex in vivo. In this report, we find that this normalization can introduce dynamic biases. In particular, the DeltaF/F is influenced by dye staining quality, and the ratio is also unstable over the course of experiments. As methods to record and analyze optical imaging signals become more precise, such biases can have an increasingly pernicious impact on the accuracy of findings, especially in the comparison of cytoarchitechtonic areas, in area-of-activation measurements, and in plasticity or developmental experiments. These dynamic biases of the DeltaF/F method may, to an extent, be mitigated by a novel method of normalization, DeltaF/DeltaF(epileptiform). This normalization uses as a reference the measured activity of epileptiform spikes elicited by global disinhibition with bicuculline methiodide. Since this normalization is based on a functional measure, i.e. the signal amplitude of "hypersynchronized" bursts of activity in the cortical network, it is less influenced by staining of non-functional elements. We demonstrate that such a functional measure can better represent the amplitude of population mass action, and discuss alternative functional normalizations based on the amplitude of synchronized spontaneous sleep-like activity. These findings demonstrate that the traditional DeltaF/F normalization of voltage-sensitive dye signals can introduce pernicious inaccuracies in the quantification of neural population activity. They further suggest that normalization-independent metrics such as waveform propagation patterns, oscillations in single detectors, and phase relationships between detector pairs may better capture the biological information which is obtained by high-sensitivity imaging.

Methods for voltage-sensitive dye imaging of rat cortical activity with high signal-to-noise ratio.

We describe methods to achieve high sensitivity in voltage-sensitive dye (VSD) imaging from rat barrel and visual cortices in vivo with the use of a blue dye RH1691 and a high dynamic range imaging device (photodiode array). With an improved staining protocol and an off-line procedure to remove pulsation artifact, the sensitivity of VSD recording is comparable with that of local field potential recording from the same location. With this sensitivity, one can record from approximately 500 individual detectors, each covering an area of cortical tissue 160 microm in diameter (total imaging field approximately 4 mm in diameter) and a temporal resolution of 1,600 frames/s, without multiple-trial averaging. We can record 80-100 trials of intermittent 10-s trials from each imaging field before the VSD signal reduces to one half of its initial amplitude because of bleaching and wash-out. Taken together, the methods described in this report provide a useful tool for visualizing evoked and spontaneous waves from rodent cortex.

Expression of distinct alpha subunits of GABAA receptor regulates inhibitory synaptic strength.

Distinct alpha subunit subtypes in the molecular assembly of GABA(A) receptors are a critical determinant of the functional properties of inhibitory synapses and their modulation by a range of pharmacological agents. We investigated the contribution of these subunits to the developmental changes of inhibitory synapses in cerebellar granule neurons in primary cultures from wild-type and alpha1 subunit -/- mice. The decay time of miniature inhibitory postsynaptic currents (mIPSCs) halved between 6 days in vitro (DIV6) and DIV12. This was paralleled by the decrease of alpha2 and alpha3 subunits, the increase of alpha1 and alpha6 subunits expression at synapses, and changes in the action of selective alpha subunit modulators. A small but significant shortening of mIPSCs was observed with development in cells from -/- mice together with a decrease in the expression of alpha3 subunit. In contrast, the expression of alpha2 subunit at inhibitory synapses in -/- cells was significantly higher than in +/+ cells at DIV11-12. alpha5 subunit was not detected, and increased sensitivity to a selective alpha4/alpha6 subunit agonist suggests increased expression of extrasynaptic receptors in -/- mice. beta2/beta3 subunit expression and loreclezole sensitivity increased with development in +/+ but not in -/- cells, supporting the preferential association of the alpha1 with the beta2 subunit. Synaptic charge transfer strongly decreased with development but was not different between cells in the +/+ and -/- groups until DIV11-12. Our results uncover a pattern of sequential expression of alpha subunits underlying the changes in functional efficacy of GABAergic networks with development.