Ebolavirus causes sporadic outbreaks of lethal hemorrhagic fever in humans with no currently approved therapy. Cells take up Ebolavirus by macropinocytosis, followed by trafficking through endosomal vesicles. However, few factors controlling endosomal virus movement are known. Here we find that Ebolavirus entry into host cells requires the endosomal calcium channels called two pore channels (TPCs). Disrupting TPC function by gene knockout, small interfering RNAs or small molecule inhibitors halted virus trafficking and prevented infection. Tetrandrine, the most potent small molecule we tested, inhibited infection of human macrophages, the primary target of Ebolavirus in vivo, and also showed therapeutic efficacy in mice. Therefore, TPC proteins play a key role in Ebolavirus infection and may be effective targets for antiviral therapy.

Although the hippocampus is critical to episodic memory, neuronal representations supporting this role, especially relating to nonspatial information, remain elusive. Here, we investigated rate and temporal coding of hippocampal CA1 neurons in rats performing a cue-combination task that requires the integration of sequentially provided sound and odor cues. The majority of CA1 neurons displayed sensory cue-, combination-, or choice-specific (simply, "event"-specific) elevated discharge activities, which were sustained throughout the event period. These event cells underwent transient theta phase precession at event onset, followed by sustained phase locking to the early theta phases. As a result of this unique single neuron behavior, the theta sequences of CA1 cell assemblies of the event sequences had discrete representations. These results help to update the conceptual framework for space encoding toward a more general model of episodic event representations in the hippocampus.

BackgroundFor most classes of drugs, rapid development of therapeutics to treat emerging infections is challenged by the timelines needed to identify compounds with the desired efficacy, safety, and pharmacokinetic profiles. Fully human monoclonal antibodies (mAbs) provide an attractive method to overcome many of these hurdles to rapidly produce therapeutics for emerging diseases.MethodsIn this study, we deployed a platform to generate, test, and develop fully human antibodies to Zaire ebolavirus. We obtained specific anti-Ebola virus (EBOV) antibodies by immunizing VelocImmune mice that use human immunoglobulin variable regions in their humoral responses.ResultsOf the antibody clones isolated, 3 were selected as best at neutralizing EBOV and triggering FcγRIIIa. Binding studies and negative-stain electron microscopy revealed that the 3 selected antibodies bind to non-overlapping epitopes, including a potentially new protective epitope not targeted by other antibody-based treatments. When combined, a single dose of a cocktail of the 3 antibodies protected nonhuman primates (NHPs) from EBOV disease even after disease symptoms were apparent.ConclusionsThis antibody cocktail provides complementary mechanisms of actions, incorporates novel specificities, and demonstrates high-level postexposure protection from lethal EBOV disease in NHPs. It is now undergoing testing in normal healthy volunteers in preparation for potential use in future Ebola epidemics.

The architectonic subdivisions of the brain are believed to be functional modules, each processing parts of global functions. Previously, we showed that neurons in different regions operate in different firing regimes in monkeys. It is possible that firing regimes reflect differences in underlying information processing, and consequently the firing regimes in homologous regions across animal species might be similar. We analyzed neuronal spike trains recorded from behaving mice, rats, cats, and monkeys. The firing regularity differed systematically, with differences across regions in one species being greater than the differences in similar areas across species. Neuronal firing was consistently most regular in motor areas, nearly random in visual and prefrontal/medial prefrontal cortical areas, and bursting in the hippocampus in all animals examined. This suggests that firing regularity (or irregularity) plays a key role in neural computation in each functional subdivision, depending on the types of information being carried.Key words: firing irregularity/regularity; interspecies similarity; neuronal firing pattern; neuronal firing regime Significance StatementBy analyzing neuronal spike trains recorded from mice, rats, cats, and monkeys, we found that different brain regions have intrinsically different firing regimes that are more similar in homologous areas across species than across areas in one species. Because different regions in the brain are specialized for different functions, the present finding suggests that the different activity regimes of neurons are important for supporting different functions, so that appropriate neuronal codes can be used for different modalities.

Ebola virus outbreaks, such as the 2014 Makona epidemic in West Africa, are episodic and deadly. Filovirus antivirals are currently not clinically available. Our findings suggest interferon gamma, an FDA-approved drug, may serve as a novel and effective prophylactic or treatment option. Using mouse-adapted Ebola virus, we found that murine interferon gamma administered 24 hours before or after infection robustly protects lethally-challenged mice and reduces morbidity and serum viral titers. Furthermore, we demonstrated that interferon gamma profoundly inhibits Ebola virus infection of macrophages, an early cellular target of infection. As early as six hours following in vitro infection, Ebola virus RNA levels in interferon gamma-treated macrophages were lower than in infected, untreated cells. Addition of the protein synthesis inhibitor, cycloheximide, to interferon gamma-treated macrophages did not further reduce viral RNA levels, suggesting that interferon gamma blocks life cycle events that require protein synthesis such as virus replication. Microarray studies with interferon gamma-treated human macrophages identified more than 160 interferon-stimulated genes. Ectopic expression of a select group of these genes inhibited Ebola virus infection. These studies provide new potential avenues for antiviral targeting as these genes that have not previously appreciated to inhibit negative strand RNA viruses and specifically Ebola virus infection. As treatment of interferon gamma robustly protects mice from lethal Ebola virus infection, we propose that interferon gamma should be further evaluated for its efficacy as a prophylactic and/or therapeutic strategy against filoviruses. Use of this FDA-approved drug could rapidly be deployed during future outbreaks.

No abstract

Multi-neuronal recording is a powerful electrophysiological technique that has revealed much of what is known about the neuronal interactions in the brain. However, it is difficult to detect precise spike timings, especially synchronized simultaneous firings, among closely neighboring neurons recorded by one common electrode because spike waveforms overlap on the electrode when two or more neurons fire simultaneously. In addition, the non-Gaussian variability (nonstationarity) of spike waveforms, typically seen in the presence of so-called complex spikes, limits the ability to sort multi-neuronal activities into their single-neuron components. Because of these problems, the ordinary spike-sorting techniques often give inaccurate results. Our previous study has shown that independent component analysis (ICA) can solve these problems and separate single-neuron components from multi-neuronal recordings. The ICA has, however, one serious limitation that the number of separated neurons must be less than the number of electrodes. The present study combines the ICA and the efficiency of the ordinary spike-sorting technique (k-means clustering) to solve the spike-overlapping and the nonstationarity problems with no limitation on the number of single neurons to be separated. First, multi-neuronal activities are sorted into an overly large number of clusters by k-means clustering. Second, the sorted clusters are decomposed by ICA. Third, the decomposed clusters are progressively aggregated into a minimal set of putative single neurons based on similarities of basis vectors estimated by ICA. We applied the present procedure to multi-neuronal waveforms recorded with tetrodes composed of four microwires in the prefrontal cortex of awake behaving monkeys. The results demonstrate that there are functional connections among neighboring pyramidal neurons, some of which fire in a precise simultaneous manner and that precisely time-locked monosynaptic connections are working between neighboring pyramidal neurons and interneurons. Detection of these phenomena suggests that the present procedure can sort multi-neuronal activities, which include overlapping spikes and realistic non-Gaussian variability of spike waveforms, into their single-neuron components. We processed several types of synthesized data sets in this procedure and confirmed that the procedure was highly reliable and stable. The present method provides insights into the local circuit bases of excitatory and inhibitory interactions among neighboring neurons.

Synchronized firing among neurons in the working brain is inferred to reflect coding by cell assemblies, which dynamically change their sizes and functional connections to encode various information. It therefore follows that, if synchronized firing reflects cell-assembly coding, it should show dynamic changes that depend on the tasks and events being processed and on the distance between the neurons. By using unique spike-sorting and multi-neuronal recording methods, we investigated such dynamics of synchrony in the prefrontal cortex of monkeys while they were successively performing two tasks in which working memory for either stimulus duration or color was required. Forty-eight percent of 1405 neuronal pairs showed firing synchrony during the performance of the tasks. Almost half of such neuronal pairs showed fixed synchrony and constantly fired together in both tasks. However, some neuronal pairs showed taskdependent synchrony that appeared in only one of the tasks. Moreover, the other neuronal pairs showed event-task-dependent synchrony that appeared during stimulus or retention periods in the tasks, but the periods showing synchrony varied between the tasks. Fixed synchrony and task-dependent synchrony were mostly observed among neighboring neurons and showed little variation of spike timings; the event-task-dependent synchrony, in contrast, was more often detected among distant neurons with larger variation of spike timings than the other two types of synchrony. These results suggest that some closely neighboring neurons have dynamic and sharp synchrony to represent certain situations (tasks), whereas some distant neurons show more dynamic and unstable synchronous firing to represent quickly changing events being processed in working memory.