Ventral Hippocampal Commissure Research Articles

Disruption of awake sharp-wave ripples does not affect memorization of locations in repeated-acquisition spatial memory tasks.

Storing and accessing memories is required to successfully perform day-to-day tasks, for example for engaging in a meaningful conversation. Previous studies in both rodents and primates have correlated hippocampal cellular activity with behavioral expression of memory. A key role has been attributed to awake hippocampal replay - a sequential reactivation of neurons representing a trajectory through space. However, it is unclear if awake replay impacts immediate future behavior, gradually creates and stabilizes long-term memories over a long period of time (hours and longer), or enables the temporary memorization of relevant events at an intermediate time scale (seconds to minutes). In this study, we aimed to address the uncertainty around the timeframe of impact of awake replay by collecting causal evidence from behaving rats. We detected and disrupted sharp wave ripples (SWRs) - signatures of putative replay events - using electrical stimulation of the ventral hippocampal commissure in rats that were trained on three different spatial memory tasks. In each task, rats were required to memorize a new set of locations in each trial or each daily session. Interestingly, the rats performed equally well with or without SWR disruptions. These data suggest that awake SWRs - and potentially replay - does not affect the immediate behavior nor the temporary memorization of relevant events at a short timescale that are required to successfully perform the spatial tasks. Based on these results, we hypothesize that the impact of awake replay on memory and behavior is long-term and cumulative over time.

Structural interhemispheric connectivity defects in mouse models of BBSOAS: Insights from high spatial resolution 3D white matter tractography

White matter (WM) tract formation and axonal pathfinding are major processes in brain development allowing to establish precise connections between targeted structures. Disruptions in axon pathfinding and connectivity impairments will lead to neural circuitry abnormalities, often associated with various neurodevelopmental disorders (NDDs). Among several neuroimaging methodologies, Diffusion Tensor Imaging (DTI) is a magnetic resonance imaging (MRI) technique that has the advantage of visualizing in 3D the WM tractography of the whole brain non-invasively. DTI is particularly valuable in unpinning structural tract connectivity defects of neural networks in NDDs. In this study, we used 3D DTI to unveil brain-specific tract defects in two mouse models lacking the Nr2f1 gene, which mutations in patients have been proven to cause an emerging NDD, called Bosch-Boonstra-Schaaf Optic Atrophy (BBSOAS). We aimed to investigate the impact of the lack of cortical Nr2f1 function on WM morphometry and tract microstructure quantifications. We found in both mutant mice partial loss of fibers and severe misrouting of the two major cortical commissural tracts, the corpus callosum, and the anterior commissure, as well as the two major hippocampal efferent tracts, the post-commissural fornix, and the ventral hippocampal commissure. DTI tract malformations were supported by 2D histology, 3D fluorescent imaging, and behavioral analyses. We propose that these interhemispheric connectivity impairments are consistent in explaining some cognitive defects described in BBSOAS patients, particularly altered information processing between the two brain hemispheres. Finally, our results highlight 3DDTI as a relevant neuroimaging modality that can provide appropriate morphometric biomarkers for further diagnosis of BBSOAS patients.

Spatial and amplitude dynamics of neurostimulation: Insights from the acute intrahippocampal kainate seizure mouse model.

Neurostimulation is an emerging treatment for patients with drug-resistant epilepsy, which is used to suppress, prevent, and terminate seizure activity. Unfortunately, after implantation and despite best clinical practice, most patients continue to have persistent seizures even after years of empirical optimization. The objective of this study is to determine optimal spatial and amplitude properties of neurostimulation in inhibiting epileptiform activity in an acute hippocampal seizure model. We performed high-throughput testing of high-frequency focal brain stimulation in the acute intrahippocampal kainic acid mouse model of status epilepticus. We evaluated combinations of six anatomic targets and three stimulus amplitudes. We found that the spike-suppressive effects of high-frequency neurostimulation are highly dependent on the stimulation amplitude and location, with higher amplitude stimulation being significantly more effective. Epileptiform spiking activity was significantly reduced with ipsilateral 250 μA stimulation of the CA1 and CA3 hippocampal regions with 21.5% and 22.2% reductions, respectively. In contrast, we found that spiking frequency and amplitude significantly increased with stimulation of the ventral hippocampal commissure. We further found spatial differences with broader effects from CA1 versus CA3 stimulation. These findings demonstrate that the effects of therapeutic neurostimulation in an acute hippocampal seizure model are highly dependent on the location of stimulation and stimulus amplitude. We provide a platform to optimize the anti-seizure effects of neurostimulation, and demonstrate that an exploration of the large electrical parameter and location space can improve current modalities for treating epilepsy. In this study, we tested how electrical pulses in the brain can help control seizures in mice. We found that the electrode's placement and the stimulation amplitude had a large effect on outcomes. Some brain regions, notably nearby CA1 and CA3, responded positively with reduced seizure-like activities, while others showed increased activity. These findings emphasize that choosing the right spot for the electrode and adjusting the strength of electrical pulses are both crucial when considering neurostimulation treatments for epilepsy.

The Induction of Long-Term Potentiation by Medial Septum Activation under Urethane Anesthesia Can Alter Gene Expression in the Hippocampus.

We studied changes in the expression of early genes in hippocampal cells in response to stimulation of the dorsal medial septal area (dMSA), leading to long-term potentiation in the hippocampus. Rats under urethane anesthesia were implanted with stimulating electrodes in the ventral hippocampal commissure and dMSA and a recording electrode in the CA1 area of the hippocampus. We found that high-frequency stimulation (HFS) of the dMSA led to the induction of long-term potentiation in the synapses formed by the ventral hippocampal commissure on the hippocampal CA1 neurons. One hour after dMSA HFS, we collected the dorsal and ventral hippocampi on both the ipsilateral (damaged by the implanted electrode) and contralateral (intact) sides and analyzed the expression of genes by qPCR. The dMSA HFS led to an increase in the expression of bdnf and cyr61 in the ipsilateral hippocampi and egr1 in the ventral contralateral hippocampus. Thus, dMSA HFS under the conditions of degeneration of the cholinergic neurons in the medial septal area prevented the described increase in gene expression. The changes in cyr61 expression appeared to be dependent on the muscarinic M1 receptors. Our data suggest that the induction of long-term potentiation by dMSA activation enhances the expression of select early genes in the hippocampus.

CA3-CA1 long-term potentiation occurs regardless of respiration and cardiac cycle phases in urethane-anesthetized rats.

Breathing and heartbeat synchronize to each other and to brain function and affect cognition in humans. However, it is not clear how cardiorespiratory rhythms modulate such basic processes as synaptic plasticity thought to underlie learning. Thus, we studied if respiration and cardiac cycle phases at burst stimulation onset affect hippocampal long-term potentiation (LTP) in the CA3-CA1 synapse in urethane-anesthetized adult male Sprague-Dawley rats. In a between-subjects design, we timed burst stimulation of the ventral hippocampal commissure (vHC) to systole or diastole either during expiration or inspiration and recorded responses throughout the hippocampus with a linear probe. As classical conditioning in humans seems to be most efficient at expiration-diastole, we also expected LTP to be most efficient if burst stimulation was targeted to expiration-diastole. However, LTP was induced equally in all four groups and respiration and cardiac cycle phase did not modulate CA1 responses to vHC stimulation overall. This could be perhaps because we bypassed all natural routes of external influences on the CA1 by directly stimulating the vHC. In the future, the effect of cardiorespiratory rhythms on synaptic plasticity could also be studied in awake state and in other parts of the hippocampal tri-synaptic loop.

Simultaneous Cellular Imaging, Electrical Recording and Stimulation of Hippocampal Activity in Freely Behaving Mice.

Hippocampal sharp-wave ripple activity (SWRs) and the associated replay of neural activity patterns are well-known for their role in memory consolidation. This activity has been studied using electrophysiological approaches, as high temporal resolution is required to recognize SWRs in the neuronal signals. However, it has been difficult to analyze the individual contribution of neurons to task-specific SWRs, because it is hard to track neurons across a long time with electrophysiological recording. In this study, we recorded local field potential (LFP) signals in the hippocampal CA1 of freely behaving mice and simultaneously imaged calcium signals in contralateral CA1 to leverage the advantages of both electrophysiological and imaging approaches. We manufactured a custom-designed microdrive array and targeted tetrodes to the left hippocampus CA1 for LFP recording and applied electrical stimulation in the ventral hippocampal commissure (VHC) for closed-loop disruption of SWRs. Neuronal population imaging in the right hippocampal CA1 was performed using a miniature fluorescent microscope (Miniscope) and a genetically encoded calcium indicator. As SWRs show highly synchronized bilateral occurrence, calcium signals of SWR-participating neurons could be identified and tracked in spontaneous or SWR-disrupted conditions. Using this approach, we identified a subpopulation of CA1 neurons showing synchronous calcium elevation to SWRs. Our results showed that SWR-related calcium transients are more disrupted by electrical stimulation than non-SWR-related calcium transients, validating the capability of the system to detect and disrupt SWRs. Our dual recording method can be used to uncover the dynamic participation of individual neurons in SWRs and replay over extended time windows.

Transection of the ventral hippocampal commissure impairs spatial reference but not contextual or spatial working memory.

Plasticity is a neural phenomenon in which experience induces long-lasting changes to neuronal circuits and is at the center of most neurobiological theories of learning and memory. However, too much plasticity is maladaptive and must be balanced with substrate stability. Area CA3 of the hippocampus provides such a balance via hemispheric lateralization, with the left hemisphere dominant in providing plasticity and the right specialized for stability. Left and right CA3 project bilaterally to CA1; however, it is not known whether this downstream merging of lateralized plasticity and stability is functional. We hypothesized that interhemispheric convergence of input from these pathways is essential for integrating spatial memory stored in the left CA3 with navigational working memory facilitated by the right CA3. To test this, we severed interhemispheric connections between the left and right hippocampi in mice and assessed learning and memory. Despite damage to this major hippocampal fiber tract, hippocampus-dependent navigational working memory and short- and long-term memory were both spared. However, tasks that required the integration of information retrieved from memory with ongoing navigational working memory and navigation were impaired. We propose that one function of interhemispheric communication in the mouse hippocampus is to integrate lateralized processing of plastic and stable circuits to facilitate memory-guided spatial navigation.

P2. Unsupervised anomaly detection for diagnosing brain disorders from EEG recordings – Results from a rodent epilepsy model

We present a general machine learning (ML) framework for diagnosing brain disorders using EEG recordings. It could detect any systematic deviations from the baseline (BL) EEG recordings. We test the approach using a rodent model of an acquired mesial temporal lobe epilepsy (mTLE) via electrical perforant pathway stimulation (PPS) [1] followed by usually a latent period of several weeks of epileptogenesis (EPG) progression without any seizures. Identifying ongoing EPG during this latent period prior to the first spontaneous seizure might enable early intervention. Previous supervised approaches have proven that deep neural networks can distinguish intracranial EEG signals between the BL period prior to PPS and the EPG period after PPS [2]. Here, we formulate the problem as an unsupervised anomaly detection task, which allows learning the statistics of the input data without the need for any labels as in supervised approaches. Specifically, we first train an autoencoder (AE) to learn to encode EEG data from the BL period. We then apply the trained AE on data from the EPG period. We expect that activity patterns systematically deviating from those observed during BL should be characterized by higher reconstruction errors (REs). Indeed, our results show that REs averaged over one hour gradually increase as a function of time after the PPS (Fig. 1). We also find that the distribution of REs from the BL and the EPG phases are significantly different (Fig. 1B). EEG segments with particularly high REs exhibit spikes and high frequency oscillations (Fig. 1B, right inset). Interestingly, there are an increased large number of segments with unusually low REs (Fig. 1B, left inset), which exhibits EEG slowing and delta rhythm (1.5–3 Hz). In conclusion, we showed that unsupervised ML with AEs allows the automatic detection of systematic drifts in brain activity, which may facilitate rapid EEG recordings for early diagnosis of brain disorders and open the door for early interventions. References Costard, L.S., Neubert, V., Venø, M.T., Su, J., Kjems, J., Connolly, N.M., Prehn, J.H., Schratt, G., Henshall, D.C., Rosenow, F. and Bauer, S., 2019. Electrical stimulation of the ventral hippocampal commissure delays experimental epilepsy and is associated with altered microRNA expression. Brain Stimulation, 12(6), pp. 1390-1401. Lu, D., Bauer, S., Neubert, V., Costard, L.S., Rosenow, F. and Triesch, J., 2020, September. Towards Early Diagnosis of Epilepsy from EEG Data. In Machine Learning for Healthcare Conference (pp. 80-96). PMLR. Fig. 1. A. The reconstruction error evolution throughout the whole recording period of rat #1B. The distribution of the reconstruction errors from the BL and the EPG phases from the recordings of rat #1. We also show some typical EPG segments whose reconstruction errors are below the 10th percentile and over the 90th percentile, highlighted with a solid and a dashed black box, respectively. Other stimulated rats show a similar trend.

Long-term potentiation in the hippocampal CA3 to CA1 synapses may be induced in vivo by activation of septal cholinergic inputs

Purpose/aim The role of cholinergic neurotransmission in the hippocampus remains controversial since different studies showed either no influence or its modulatory effect on glutamatergic hippocampal synapses. It remains unclear whether septal cholinergic input can modulate plasticity of synapses formed by CA3 pyramids on CA1 neurons. The aim of the study was to clarify the role of septal input in the development of LTP in this synapse. Materials and methods We recorded in vivo in rats under urethane anesthesia focal excitatory postsynaptic potential (fEPSP) characteristics in CA1 area after stimulation of the ventral hippocampal commissure (VHC), which contains both CA3 axons innervating CA1 neurons and cholinergic axons coming from the medial septum. We performed two series of experiments in which LTP was induced by tetanization of either VHC or medial septal area (MSA). Degeneration of cholinergic neurons in MSA was induced by intraseptal injection of 192IgG-saporin. Results In both experimental series, tetanization induced an increase in fEPSP amplitude which lasted for at least 40 min after tetanic stimulation, although tetanization of VHC induced a larger increase in fEPSP amplitude compared to MSA tetanization. Elimination of septal cholinergic neurons by 192IgG-saporin abolished LTP development in both experimental series. This suppression of LTP in animals with cholinergic deficit was not due to loss of hippocampal neurons. Conclusions Our data suggest that activation of septal cholinergic fibers during tetanization is a critical factor of LTP induction in the hippocampal CA3 to CA1 synapses.

Diffusion MRI revealed altered inter-hippocampal projections in the mouse brain after intrauterine inflammation.

Diffusion MRI (dMRI) is commonly used to map large axonal pathways in the white matter. Recent technical advances have also enabled dMRI to resolve the small and complex axonal and dendritic projections in the gray matter. This study investigated whether high-resolution dMRI can resolve the hippocampal neuronal projections and detect abnormal connections due to neurological injury. We performed 3D high spatial and angular resolution dMRI of the mouse brains of the offspring survivors from a model of intrauterine (UI) inflammation, who had known functional deficiency in the hippocampus. We used a novel hippocampal connection mapping method to quantify the intra- and inter-hippocampal projections among 34 automatically segmented hippocampal sub-regions. The results demonstrated wide-spread intra-hippocampal projections, but rather specific intra-hippocampal projections that primarily connected through the CA3 region. Compared with the control group (n = 9), UI-injured mice (n = 11) exhibited significantly reduced inter-hippocampal projection strength (p < 0.01), which correlated well with the neurobehavioral assessments (R2 = 0.47). Furthermore, using a whole-brain fixel-based analysis, we identified reduced fiber-density in the CA3 and the ventral hippocampal commissure of the UI-injured mice, which may explain the reduced inter-hippocampal projections. Histological findings also indicated reduced commissural fibers due to the UI-injury. Our study suggested that the dMRI-based connectivity mapping technique can potentially characterize abnormal hippocampal projections in neurological disorders.

Electrical stimulation of the ventral hippocampal commissure delays experimental epilepsy and is associated with altered microRNA expression

BackgroundUp to 80% of mesial temporal lobe epilepsy patients with hippocampal sclerosis (mTLE-HS) are resistant to pharmacological treatment, often necessitating surgical resection. Deep brain stimulation (DBS) has emerged as an alternative treatment for patients who do not qualify for resective brain surgery. Brain stimulation may also exert disease-modifying effects, and noncoding microRNAs have recently been proposed to shape the gene expression landscape in epilepsy. ObjectiveWe compared the effect of DBS of 4 different hippocampal target regions on epileptogenesis and manifest epilepsy in a rat model of mTLE-HS. To explore mechanisms, we profiled the effect of the most effective DBS paradigm on hippocampal microRNA levels. MethodsMTLE-HS was induced by electrical stimulation of the perforant pathway (PP) in rats. This paradigm leads to spontaneous seizures within 4 weeks. We investigated DBS of 4 targets: PP, fimbria fornix (FF) formation, dentate gyrus (DG) and ventral hippocampal commissure (VHC). We applied both high- (130 Hz) and low-frequency (5 Hz or 1 Hz) stimulation. Functional microRNAs were identified in the hippocampus immediately after VHC-DBS and after a 97-day recording period by sequencing small RNAs bound to Argonaute-2, a component of the miRNA silencing complex. ResultsLow frequency DBS of the VHC significantly delayed the occurrence of the first spontaneous recurrent seizure in the PPS model by ∼300%, from 19 to 56 days. No other stimulation regime altered the latency phase. Upregulation of 5 microRNAs during epileptogenesis was suppressed by VHC-stimulation. ConclusionWe conclude that DBS of the VHC delays epilepsy in the PPS model in rats and is associated with differential regulation of several miRNAs. Additional studies are required to determine whether VHC-regulated miRNAs serve causal roles in the anti-epileptogenic effects of this DBS model.

Trajectory of hippocampal fibres to the contralateral anterior thalamus and mammillary bodies in rats, mice, and macaque monkeys.

The routes by which the hippocampal formation projects bilaterally to the anterior thalamic nuclei and mammillary bodies were examined in the mouse, rat, and macaque monkey. Despite using different methods and different species, the principal pattern remained the same. For both target areas, the contralateral hippocampal (subiculum) projections arose via efferents in the postcommissural fornix ipsilateral to the tracer injection, which then crossed hemispheres both in or just prior to reaching the target site within the thalamus or hypothalamus. Precommissural fornix fibres could not be followed to the target areas. There was scant evidence that the ventral hippocampal commissure or decussating fornix fibres contribute to these crossed subiculum projections. Meanwhile, a small minority of postsubiculum projections in the mouse were seen to cross in the descending fornix at the level of the caudal septum to join the contralateral postcommissural fornix before reaching the anterior thalamus and lateral mammillary nucleus on that side. Although the rodent anterior thalamic nuclei also receive nonfornical inputs from the subiculum and postsubiculum via the ipsilateral internal capsule, few, if any, of these projections cross the midline. It was also apparent that nuclei within the head direction system (anterodorsal thalamic nucleus, laterodorsal thalamic nucleus, and lateral mammillary nucleus) receive far fewer crossed hippocampal inputs than the other anterior thalamic or mammillary nuclei. The present findings increase our understanding of the fornix and its component pathways while also informing disconnection analyses involving the hippocampal formation and diencephalon.

Dentate spikes and learning: disrupting hippocampal function during memory consolidation can improve pattern separation.

Hippocampal dentate spikes (DSs) are short-duration, large-amplitude fluctuations in hilar local field potentials and take place while resting and sleeping. During DSs, dentate gyrus granule cells increase firing while CA1 pyramidal cells decrease firing. Recent findings suggest DSs play a significant role in memory consolidation after training on a hippocampus-dependent, nonspatial associative learning task. Here, we aimed to find out whether DSs are important in other types of hippocampus-dependent learning tasks as well. To this end, we trained adult male Sprague-Dawley rats in a spatial reference memory task, a fixed interval task, and a pattern separation task. During a rest period immediately after each training session, we either let neural activity to take place as usual, timed electrical stimulation of the ventral hippocampal commissure (vHC) to immediately follow DSs, or applied the vHC stimulation during a random neural state. We found no effect of vHC stimulation on performance in the spatial reference memory task or in the fixed interval task. Surprisingly, vHC stimulation, especially contingent on DSs, improved performance in the pattern separation task. In conclusion, the behavioral relevance of hippocampal processing and DSs seems to depend on the task at hand. It could be that in an intact brain, offline memory consolidation by default involves associating neural representations of temporally separate but related events. In some cases this might be beneficial for adaptive behavior in the future (associative learning), while in other cases it might not (pattern separation). NEW & NOTEWORTHY The behavioral relevance of dentate spikes seems to depend on the learning task at hand. We suggest that dentate spikes are related to associating neural representations of temporally separate but related events within the dentate gyrus. In some cases this might be beneficial for adaptive behavior in the future (associative learning), while in other cases it might not (pattern separation).

P71. MicroRNA regulation in deep brain stimulation of different targets for epilepsy

Rationale Epilepsy is a common neurological disorder with an incidence of 50.4 per 100,000 per year ( Ngugi et al., 2011 ). Up to 80% of mesial temporal lobe epilepsy (mTLE) patients are resistant to pharmacological treatment ( Semah et al., 1998 ). Many of them do not qualify for resective brain surgery. For these patients, an alternative treatment is deep brain stimulation (DBS). In epilepsy, different protocols for DBS have been tested but until now the mechanisms of DBS are not fully understood and the best protocol remains to be evaluated. MicroRNAs (miRNAs) are small, single stranded RNAs, which regulate protein synthesis on mRNA level. Various miRNAs have been found to be regulated in epilepsy, and regulation of miRNAs after DBS has been shown in Parkinson’s disease patients ( Soreq et al., 2013 ). We compared different targets and frequencies of DBS in a rat model of mTLE to identify a paradigm that inhibits epileptogenesis. Furthermore, we investigated miRNA regulation after DBS with the most effective paradigm. Methods We used the rat perforant pathways stimulation (PPS) model of mTLE ( Norwood et al., 2010 ). Thereby, stimulation electrodes are implanted into the PP and recording electrodes are implanted into the dentate gyrus (DG). After one week of recovery, rats are stimulated on two days for 30 min and on the third day for 8 h. This treatment leads to spontaneous seizures 12–34 days after stimulation. We investigated DBS of four different targets: the PP, the fimbria fornix formation (FF), the DG and the ventral hippocampal commissure (VHC); for one or two weeks starting one day after the 8 h stimulation, using three different stimulation paradigms: 130 Hz and 0.1 V, 5 Hz and 2 V or 1 V and 1 Hz. We furthermore analyzed miRNA expression in the hippocampus immediately after DBS and after a 97-day recording period. Thereafter, sequencing of active miRNAs was performed and compared to control rats which were not treated with DBS. Results A two-week DBS of the VHC with 1 Hz and 1 V significantly prolonged the latency period in the PPS model from 19 to 56 days. 8 microRNAs were significantly changed directly after DBS. Several miRNAs were changed in the control group after 97 days but showed no changes in expression in the DBS-treated group. These are potential candidates for an in vivo manipulation to inhibit epileptogenesis.

The Relationship between Neurite Density Measured with Confocal Microscopy in a Cleared Mouse Brain and Metrics Obtained from Diffusion Tensor and Diffusion Kurtosis Imaging.

Purpose:Diffusional kurtosis imaging (DKI) enables sensitive measurement of tissue microstructure by quantifying the non-Gaussian diffusion of water. Although DKI is widely applied in many situations, histological correlation with DKI analysis is lacking. The purpose of this study was to determine the relationship between DKI metrics and neurite density measured using confocal microscopy of a cleared mouse brain.Methods:One thy-1 yellow fluorescent protein 16 mouse was deeply anesthetized and perfusion fixation was performed. The brain was carefully dissected out and whole-brain MRI was performed using a 7T animal MRI system. DKI and diffusion tensor imaging (DTI) data were obtained. After the MRI scan, brain sections were prepared and then cleared using aminoalcohols (CUBIC). Confocal microscopy was performed using a two-photon confocal microscope with a laser. Forty-eight ROIs were set on the caudate putamen, seven ROIs on the anterior commissure, and seven ROIs on the ventral hippocampal commissure on the confocal microscopic image and a corresponding MR image. In each ROI, histological neurite density and the metrics of DKI and DTI were calculated. The correlations between diffusion metrics and neurite density were analyzed using Pearson correlation coefficient analysis.Results:Mean kurtosis (MK) (P = 5.2 × 10−9, r = 0.73) and radial kurtosis (P = 2.3 × 10−9, r = 0.74) strongly correlated with neurite density in the caudate putamen. The correlation between fractional anisotropy (FA) and neurite density was moderate (P = 0.0030, r = 0.42). In the anterior commissure and the ventral hippocampal commissure, neurite density and FA are very strongly correlated (P = 1.3 × 10−5, r = 0.90). MK in these areas were very high value and showed no significant correlation (P = 0.48).Conclusion:DKI accurately reflected neurite density in the area with crossing fibers, potentially allowing evaluation of complex microstructures.

Hippocampal electrical stimulation disrupts associative learning when targeted at dentate spikes.

Dentate spikes are fast fluctuations of hilar local-field potentials that take place during rest and are thought to reflect input arriving from the entorhinal cortex to the hippocampus. During dentate spikes, neuronal firing in hippocampal input (dentate gyrus) and output (CA1/CA3) regions is uncoupled. To date, the behavioural significance of dentate spikes is unknown. Here, we provide evidence that disrupting the dentate spike-related uncoupling of the dentate gyrus and the CA1/CA3 subregions for 1h after training retards associative learning. We suggest dentate spikes play a significant role in memory consolidation. Hippocampal electrophysiological oscillations, namely theta and ripples, have been implicated in encoding and consolidation of new memories, respectively. According to existing literature, hippocampal dentate spikes are prominent, short-duration (<30ms), large-amplitude (∼2-4mV) fluctuations in hilar local-field potentials that take place during awake immobility and sleep. Interestingly, previous studies indicate that during dentate spikes dentate gyrus granule cells increase their firing while firing of CA1 pyramidal cells are suppressed, thus resulting in momentary uncoupling of the two hippocampal subregions. To date, the behavioural significance of dentate spikes is unknown. Here, to study the possible role of dentate spikes in learning, we trained adult male Sprague-Dawley rats in trace eyeblink classical conditioning. For 1h immediately following each conditioning session, one group of animals received hippocampal stimulation via the ventral hippocampal commissure (vHC) contingent on dentate spikes to disrupt the uncoupling between the dentate gyrus and the CA1 subregions. A yoked control group was stimulated during immobility, irrespective of brain state, and another control group was not stimulated at all. As a result, learning was impaired only in the group where vHC stimulation was administered contingent on dentate spikes. Our results suggest dentate spikes and/or the associated uncoupling of the dentate gyrus and the CA1 play a significant role in memory consolidation. Dentate spikes could possibly reflect reactivation and refinement of a memory trace within the dentate gyrus triggered by input from the entorhinal cortex.

In vivo visualization of connections among revised Papez circuit hubs using full q-space diffusion spectrum imaging tractography

Structural connections among the hubs of the revised Papez circuit remain to be elucidated in the human brain. As the original Papez circuit failed to explain functional imaging findings, a more detailed investigation is needed to delineate connections among the circuit’s key hubs. Here we acquired diffusion spectrum imaging (DSI) from eight normal subjects and used data from the Human Connectome Project (HCP) to elucidate connections among hubs in the retrosplenial gyrus, hippocampus, mammillary bodies, and anterior thalamic nuclei. Our results show that the ventral hippocampal commissure (VHC) was visualized in all eight individual DSI datasets, as well as in the DSI and HCP group datasets, but a strictly defined VHC was only visualized in one individual dataset. Thalamic fibers were observed to connect with both the posterior cingulate cortex (PCC) and retrosplenial cortex (RSC). The RSC was mainly responsible for direct hippocampal connections, while the PCC was not. This indicates that the RSC and PCC represent separate functional hubs in humans, as also shown by previous primate axonal tracing studies and functional magnetic resonance imaging observations.

Beneficial Effects of Selective Orexin-A Receptor Antagonist in 4-aminopyridine-induced Seizures in Male Rats.

Background:Orexins are excitatory neuropeptides which stimulate the central regulatory pathways. Orexins increase the penicillin-induced epileptic activity in rats. Orexin-A increases in different types of seizures and its elevated level is the characteristic feature in the epileptic children during polysomnography. Recently, the orexin receptor blockage has been reported to increase seizure threshold in mice; however, effect of the selective orexin-A receptor antagonist (SB-334867) on 4-aminopyridine (4-AP)-induced seizures has not been investigated.Materials and Methods:We used the intraperitoneal injection of 4-AP to induce seizure in male rats. Under urethane anesthesia, SB-334867 (50 and 100 nmol) was injected stereotaxically into the ventral hippocampal commissure. Using video recording, the effects of SB-334867 on electroencephalogram and tonic–clonic convulsions were compared to those that received diazepam or dimethyl sulfoxide (DMSO).Results:SB-334867 significantly decreased the duration of spike trains compared to DMSO-treated rats (P < 0.001) and reduced the duration of convulsive seizures (P < 0.05). Seizure onset was increased significantly by SB-334867, 50 nmol, compared to DMSO (P < 0.05) and diazepam (P < 0.01) treated rats.Conclusion:Antagonism of orexin-A receptor by a low-dose SB-334867 showed protective effects in 4-AP-induced seizure-like activities in anesthetized rats.

See-through Brains and Diffusion Tensor MRI Clarified Fiber Connections: A Preliminary Microstructural Study in a Mouse with Callosal Agenesis.

Clearing methods that render the brain optically transparent allow high-resolution three-dimensional (3D) imaging of neural networks. We used diffusion tensor imaging (DTI) and two-photon imaging of cleared brains to analyze white matter in BTBR mice. We confirmed corpus callosum agenesis and identified an abnormal commissure close to the third ventricle. DTI and cleared-brain two-photon imaging revealed that these commissural fibers constituted a frontal clustering of the ventral hippocampal commissure and provided a detailed assessment of white matter structure in mice.

Suppression of acute seizures by theta burst electrical stimulation of the hippocampal commissure using a closed-loop system

This study investigated the effects of electrical stimulation with theta burst stimulation (eTBS) on seizure suppression. Optimal parameters of eTBS were determined through open-loop stimulation experiments and then implemented in a close-loop seizure control system. For the experiments, 4-aminopyridine (4-AP) was injected into the right hippocampus of Sprague–Dawley rats to induce an acute seizure. eTBS was applied on the ventral hippocampal commissure and the effects of eTBS with different combinations of burst frequency and number of pulses per burst were analyzed in terms of seizure suppression. A closed-loop seizure control system was then implemented based on optimal eTBS parameters. The efficiency of the closed-loop eTBS was evaluated and compared to that of high frequency stimulation. The results show that eTBS induced global suppression in the hippocampus and this was sustained even after the application of eTBS. The optimal parameter of eTBS in the open-loop stimulation experiments was a burst frequency at 100Hz with nine pulses in a burst. The eTBS integrated with the on–off control law yielded less actions and cumulative delivered charge, but induced longer after-effects of seizure suppression compared to continuous high frequency stimulation (cHFS). To conclude, eTBS has suppressive effects on 4-AP induced seizure. A closed-loop eTBS system provides a more effective way of suppressing seizure and requires less effort compared to cHFS. eTBS may be a novel stimulation protocol for effective seizure control.