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Navarro, V.

Publications and source records attributed to Navarro, V..

14 recordsLinked to original sources

A learning-evoked slow-oscillatory architecture paces population activity for offline reactivation across the human medial temporal lobe

Memory processing requires the coordinated engagement of neuronal populations across distributed brain networks and across time. How such coordination is organized in the human medial temporal lobe (MTL) remains unclear. Here, we show that MTL population activity is dynamically structured by a transient slow-oscillatory architecture that emerges during online learning to promote offline consolidation and later recall. Using intracranial recordings that combine single-neuron spiking activity and local field potentials in human participants, we find that mnemonic engagement evokes on-demand slow-oscillatory bursts in the hippocampus. These hippocampal bursts synchronize gamma-band patterns across MTL regions, defining discrete coordination events that pace cross-regional coactivity motifs during learning. These learning-time population motifs are then selectively reactivated during hippocampal ripples in post-learning rest, and the strength of their reactivation predicts subsequent recall accuracy. Together, these findings identify a multi-scale coordination mechanism that links distributed population activity across learning, consolidation, and recall in humans.

neuroscience↗

Activating Ras-MAPK pathway variants drive hippocampal clonal competition in human epilepsy

Mesial (a.k.a., medial) temporal lobe epilepsy (MTLE) is the most common focal epilepsy1,2 and, in drug-resistant cases, is treated by surgical removal of the anterior temporal lobe, which often shows neuronal loss and gliosis consistent with hippocampal sclerosis (HS)2. MTLE with HS has minimal contribution from germline genetic variation3, and is associated with prior precipitating insults such as prolonged childhood seizures and head trauma4-6. Somatic variants in Ras-MAPK pathway genes were recently reported in a few MTLE surgical specimens7,8, but their prevalence, clinical relevance, and underlying biological mechanisms remain unknown. Targeted duplex sequencing of hippocampal DNA from 462 surgical resections revealed significant enrichment of deleterious somatic variants in MTLE versus controls, with >40% of MTLE specimens harboring activating Ras-MAPK variants in PTPN11, NF1, BRAF, KRAS, and twelve genes not previously associated with focal epilepsy. Eight Ras-MAPK genes showed positive clonal selection in MTLE. Increased somatic variant burden predicted worse surgical outcome. Somatic Ras-MAPK variants at ultra-low (<0.5%) variant allele fractions were associated with older seizure onset and HS pathology, supporting a late prenatal or postnatal origin. Ras-MAPK variants in MTLE were enriched in cells derived from hippocampal progenitors--neurons, astrocytes, oligodendrocytes--in line with the known neuronal hyperexcitability and seizures induced by Ras-MAPK overactivation9,10; in contrast, Alzheimer disease hippocampi exhibited microglial enrichment of Ras-MAPK variants, consistent with prior reports11. Single-nucleus RNA sequencing showed increased expression of Ras-MAPK genes in neurons and upregulation of pathways mediating neurogenesis and neural development in MTLE. Functional validation of novel, recurrent PTPN11 variants confirmed gain-of-function, while cellular modeling in induced pluripotent stem cells demonstrated proliferative/survival advantages for mutant cells in mosaic culture. Overall, our data suggest that somatic Ras-MAPK variants and acquired risk factors may converge on clonal competition in the hippocampus to modulate epilepsy risk.

genetics↗

Circadian rhythms in sexual behavior and their influence on reproductive outcomes in mice

Circadian clocks coordinate mammalian reproductive physiology, and circadian misalignment (resulting from shift work, jet lag, etc.) is known to impair fertility. Despite the well-established links between clock function and reproductive success, it remains unclear whether male and female mice maintained under standard ad libitum feeding conditions exhibit circadian rhythms in the propensity for sexual behavior, and to what degree such timing influences reproductive outcomes. Using standardized mating paradigms in C57BL/6J mice, we identified a circadian rhythm in sexual behavior in both sexes, with peak sexual activity most often occurring at circadian time (CT) 13-16 and a trough at CT4-7. To test the functional significance of these rhythms, we conducted 1-hour mating trials across four cohorts of C57BL/6J mice with pairs of mice having either aligned (e.g. male CT16 peak and female CT16 peak) or misaligned (e.g. male CT16 peak and female CT4 nadir) sexual behavior phases and monitored mating outcomes via ultrasound. While pregnancies were almost as frequent across all four cohorts, the numbers of live offspring were significantly more frequent when both the male and female mated at their peak phases than when both mice mated at their troughs. Notably, mating specifically during the males behavioral peak increased the likelihood of successful delivery of pups, suggesting that male circadian phase is a key determinant of miscarriage vs successful birth. These findings establish a circadian rhythm in the propensity for sexual behavior under standard feeding and housing conditions and indicate that mating time--particularly relative to the males circadian peak--can influence reproductive success. This insight provides a foundation for translational studies that explore whether intercourse timed to the circadian rhythms of the couple might help to increase fertility chances.

physiology↗

Representational dynamics during extinction of fear memories in the human brain

Extinction learning - the suppression of a previously acquired fear response - is critical for adaptive behavior and core for understanding the etiology and treatment of anxiety disorders. Electrophysiological studies in rodents have revealed critical roles of theta (4-12Hz) oscillations in amygdala and hippocampus during both fear learning and extinction, and engram research has shown that extinction relies on the formation of novel, highly context-dependent memory traces that suppress the initial fear memories. Whether similar processes occur in humans and how they relate to previously described neural mechanisms of episodic memory formation and retrieval remains unknown. Intracranial EEG (iEEG) recordings in epilepsy patients provide direct access to the deep brain structures of the fear and extinction network, while representational similarity analysis (RSA) allows characterizing the memory traces of specific cues and contexts. Here we combined these methods to show that amygdala theta oscillations during extinction learning signal safety rather than threat and that extinction memory traces are characterized by stable and context-specific neural representations that are coordinated across the extinction network. We further demonstrate that context specificity during extinction learning predicts the reoccurrence of fear memory traces during a subsequent test period, while reoccurrence of extinction memory traces predicts safety responses. Our results reveal the neurophysiological mechanisms and representational characteristics of context-dependent extinction learning in the human brain. In addition, they show that the mutual competition of fear and extinction memory traces provides a mechanistic basis for clinically important phenomena such as fear renewal and extinction retrieval.

neuroscience↗

A complete approach for circRNA therapeutics from purification to lyophilized delivery using novel ionizable lipids

Circular RNA (circRNA) has gained significant attention as a potential therapeutic tool due to its remarkable stability and resistance to degradation by exonucleases. However, scalable and efficient methods for purification and delivery remain critical challenges that must be addressed. In this study, we developed and evaluated an optimized affinity chromatography method using Oligo(dT) columns for the purification of circRNA, achieving high yield and purity comparing with high-performance liquid chromatography. Additionally, we investigated the in vivo efficacy of circRNA-Oligo(dT) encapsulated in lipid nanoparticles (LNPs) formulated with emerging STAAR ionizable lipids, including CP-LC-0867 and CP-LC-0729. Our results showed that LNPs formulated with CP-LC-0867 consistently produced higher protein expression compared to SM-102, with sustained luciferase activity observed over a 14-day period. Furthermore, we assessed the lyophilization potential of LNP-circRNA-Oligo(dT) using CP-LC-0729 to extend shelf life and eliminate the need for ultra-low temperature storage. Remarkably, the lyophilized LNPs exhibited no significant differences in protein expression compared to their non-lyophilized counterparts, demonstrating that lyophilization is a viable strategy for extending the storage and transport of circRNA therapies. These findings underscore the potential of optimized new ionizable lipids, improved purification strategies, and lyophilization techniques to enhance the scalability, stability, and practical application of circRNA therapies. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/620632v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@8bb7eeorg.highwire.dtl.DTLVardef@d8a1eeorg.highwire.dtl.DTLVardef@6f359forg.highwire.dtl.DTLVardef@1295d05_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

The PHD3-FOXO3 axis modulates the interferon type I response in microglia aggravating Alzheimer's disease progression

Microglia respond to Alzheimers disease (AD) with a variety of transcriptional responses. However, the regulation of specific transcriptional signatures and the contribution of each individual response to disease progression is only starting to be characterized. We have previously shown that hypoxia via hypoxia inducible factor 1 (HIF1) is a strong regulator of A{beta} plaque-associated microglia (A{beta}AM). Here, we characterize the role of HIF1-mediated transcription of Egln3, encoding for PHD3, in A{beta}AM. We show that oligomeric A{beta} treatment (oA{beta}) in vitro induces the expression of Hif1a and Egln3 in microglia, which correlates with the transcriptional activation of genes involved in the interferon type I signature (IFNS) in a PHD3-dependent manner. Mechanistically, we demonstrate FOXO3 to be an important repressor of IFNS in microglia, whose abundance decreases upon A{beta} presence, and, correspondingly, both in human single-nucleus (sn) and mouse A{beta}AM transcriptomics, FOXO3 DNA binding sites define the IFNS. FOXO3 repression of the IFNS is dependent on PHD3, with our results suggesting a physical interaction between both proteins in vitro. In vivo, loss of PHD3 correlate with abrogation of the IFNS and activation of the disease-associated microglia signature (DAM) in A{beta}AM. Transcriptional changes in microglia associate with increased microglia proximity to A{beta} plaques, augmented phagocytosis of A{beta} by microglia, reduced parenchymal levels of A{beta}, and an increase in small-sized plaques. PHD3 deficiency also reduced the A{beta} plaque-associated neuropathology and rescued behavioural deficits of an AD mouse model. Finally, we also demonstrate that microglial PHD3 overexpression during development in the absence of A{beta} pathology is sufficient to induce the IFNS and to behavioural alterations. Altogether, our data strongly indicate that the PHD3-FOXO3 axis controls the microglial IFNS in a cell autonomous manner, contributing to the progression of AD.

neuroscience↗

Computational analysis of SOD1-G93A mouse muscle biomarkers for comprehensive assessment of ALS progression

AimsTo identify potential image biomarkers of neuromuscular disease by analysing morphological and network-derived features in skeletal muscle biopsies from a murine model of amyotrophic lateral sclerosis (ALS), the SOD1G93A mouse, and wild-type (WT) controls at distinct stages of disease progression. MethodsUsing the NDICIA computational framework, we quantitatively evaluated histological differences between skeletal muscle biopsies from SOD1G93A and WT mice. The process involved the selection of a subset of features revealing these differences. A subset of discriminative features was selected to characterise these differences, and their temporal dynamics were assessed across disease stages. ResultsOur findings demonstrated that muscle pathology in the mutant model evolves from early alterations in muscle fibre arrangement, detectable at the presymptomatic stage through graph theory features, to the subsequent development of the typical morphological pattern of neurogenic atrophy at more advanced disease stages. ConclusionsOur assay identifies a neurogenic signature in mutant muscle biopsies, even when the disease was phenotypically imperceptible. KEY POINTS- NDICIA analysis detected differences between SOD1G93A and WT muscles at presymptomatic stage through the analysis of graph theory features. - Our computational tool identified different neurogenic-like traits in the ALS mouse model at all analysed stages of disease progression. - Differences between SOD1G93A and WT muscle images became more pronounced as the disease advanced. - The integration of UMAP into the upgraded NDICIA framework was validated as a robust alternative to PCA. - Muscle fibre characteristics in SOD1WT closely resembled those of WT mice.

neuroscience↗

Phase-Precession-Like Effects in the Anterior Insula Cortex during Reward Expectancy

Reward expectancy shapes behavioral performance by coordinating the allocation of cognitive resources, which incorporates the involvement of the anterior insular cortex (AIC). To investigate the electrophysiological mechanisms of the AIC during reward expectancy, we collected intracranial electroencephalographic data from epilepsy patients undergoing clinical monitoring. Subjects navigated a virtual T-maze containing rewards at three predetermined locations. We focused on the period immediately before entering the reward zone, termed the reward expectancy stage. During this stage, we identified the reward-specific brain patterns (RBPs) that were preactivated in the AIC. This pre-activation preceded the emergence of robust phase-amplitude coupling (PAC), where theta oscillations (5-7 Hz) modulated gamma activities (75-110 Hz). The PAC strength correlated positively with the pre-activation level of RBPs, implying the potential role of oscillatory coordination in amplifying preactivated reward representations. Strikingly, this PAC effect exhibited a specific temporal structure that peak gamma activity became progressively coupled to earlier theta phases as reward approached, mirroring the theta phase precession phenomenon previously observed in frontal and temporal lobes. We refer to this dynamic as phase-precession-like effect (PPLE). Moreover, subjects exhibiting PPLE in the AIC presented greater trial-by-trial improvements in response latency. Taken together, these findings shed light on the electrophysiological mechanisms of the AIC underlying reward expectancy.

neuroscience↗

Enhancing KCC2 function reduces interictal activity and prevents seizures in mesial temporal lobe epilepsy

The neuronal K/Cl cotransporter KCC2 regulates the transmembrane chloride gradient, which controls the efficacy of GABAergic signaling. In mesial temporal lobe epilepsy (mTLE) and other neurological disorders, reduced KCC2 expression or function can result in depolarizing GABA signaling, which is thought to contribute to pathological activity and seizures. Therefore, restoring chloride homeostasis represents a promising therapeutic strategy. We investigated the mechanisms and antiseizure effects of two small molecules, prochlorperazine (PCPZ) and CLP-257, that have been identified as potential KCC2 enhancers. We found that both compounds enhance KCC2 function and clustering in cortical neurons while reducing its membrane diffusion, without altering canonical regulatory phosphorylation. CLP-257 also selectively increased extrasynaptic, but not synaptic, GABAA receptor-mediated currents. Using in vitro recordings from resected brain tissue of patients with drug-resistant mTLE and in vivo recordings from a mouse model, we show that PCPZ and CLP-257 (or its prodrug CLP-290) effectively suppressed spontaneous epileptiform activity in both models. These findings reveal that PCPZ and CLP-257 act as genuine KCC2 enhancers and provide experimental evidence of the therapeutic potential of such compounds for treating drug-resistant mTLE. Significance statementA major challenge in treating epilepsy is the high percentage of patients with drug-resistant forms, like mesial temporal lobe epilepsy (mTLE). This study investigates a therapeutic strategy by targeting the neuronal KCC2 transporter, which is often dysfunctional in epilepsy. Our findings identify two compounds, prochlorperazine and CLP-257, that enhance KCC2 function by promoting its clustering on the cell membrane, a previously uncharacterized mechanism. Importantly, these compounds effectively reduce spontaneous epileptiform activity in human brain tissue from mTLE patients and significantly reduce seizure frequency in a mouse model. This work provides a critical proof-of-concept for activating KCC2 as a viable therapeutic approach for drug-resistant epilepsy.

neuroscience↗

Sleep increases firing rate modulation during interictal epileptic activities in mesial structures

Epileptic seizures and interictal epileptiform discharges (IEDs) are strongly influenced by sleep and circadian rhythms. However, human data on the effect of sleep on neuronal behavior during interictal activity have been lacking. We analyzed EEG data from epileptic patients implanted with macro and micro electrodes targeting mesial temporal structures. Sleep staging was performed on concomitantly recorded polysomnography and video-EEG. Automated IED detection identified thousands of IEDs per patient. Both the rate and amplitude of IEDs were increased with deeper stages of NREM sleep. Single unit activity (SUA) and multi-unit activity (MUA) increased their firing during the IED spike, and strongly decreased during the subsequent slow wave. These time-locked firing rate modulations were shown to increase during deeper stages of NREM sleep. Finally, during resting behaviour, neuronal firing rate, bursting rate and firing regularity were all shown to progressively decrease with deeper stages of NREM sleep.

neuroscience↗

The role of the thalamus in human reinforcement learning

Although the thalamus is supposed to be involved in reinforcement-based decision-making, there is no direct evidence regarding the involvement of this subcortical structure in humans. To fill this gap, we leveraged rare intra-thalamic electrophysiological recordings in patients and found that temporally structured thalamic oscillations encode key learning signals. Our findings also provide neural insight into the computational mechanisms of action inhibition in punishment avoidance learning.

neuroscience↗

The Interplay of Attention and Conscious Perception: Evidence from Human Intracerebral Recordings and Computational Modeling

How do attention and consciousness interact in the human brain? Rival theories of consciousness disagree on the role of fronto-parietal attentional networks in conscious perception. We recorded neural activity from 727 intracerebral contacts in 13 epileptic patients, while they detected near-threshold targets preceded by attentional cues. Unsupervised clustering revealed three patterns: (1) Attention-enhanced conscious report accompanied sustained right-hemisphere fronto-temporal activity, in networks connected by the superior longitudinal fasciculus (SLF) II-III, and late accumulation in bilateral dorso-prefrontal and right-hemisphere orbitofrontal cortex (SLF I-III). (2) Attentional reorienting affected conscious report through early, sustained activity in a right-hemisphere network (SLF III). (3) Conscious report accompanied left-hemisphere dorsolateral-prefrontal activity. Task modeling with recurrent neural networks identified specific excitatory and inhibitory interactions between attention and consciousness, and their causal contribution to conscious perception of near-threshold targets. Thus, distinct, hemisphere-asymmetric fronto-parietal networks support attentional gain and reorienting in shaping human conscious experience. One-Sentence SummaryIntracerebral recordings, tractography and modeling reveal the interaction of attention and consciousness in the human brain.

neuroscience↗

Spatiotemporal dynamics of human attention revealed byintracerebral recording

Exogenous attention, the process that makes external salient stimuli pop-out of a visual scene, is essential for survival. How attention-capturing events modulate human brain processing remains unclear. Here we show how the psychological construct of exogenous attention gradually emerges over large-scale gradients in the human cortex, by analyzing activity from 1,403 intracortical contacts implanted in 28 individuals, while they performed an exogenous attention task. The timing, location and task-relevance of attentional events defined a spatiotemporal gradient of three neural clusters, which mapped onto cortical gradients and presented a hierarchy of timescales. Visual attributes modulated neural activity at one end of the gradient, while at the other end it reflected the upcoming response timing, with attentional effects occurring at the intersection of visual and response signals. These findings challenge multi-step models of attention, and suggest that frontoparietal networks, which process sequential stimuli as separate events sharing the same location, drive exogenous attention phenomena such as inhibition of return.

neuroscience↗

Micro-electrode in vivo signatures of human periventricular heterotopia

Periventricular nodular heterotopia (PNH) is a malformation of cortical development that frequently causes drug-resistant epilepsy. The epileptogenicity of ectopic neurons in PNH as well as their role in generating interictal and ictal activity is still a matter of debate. We report the first in vivo microelectrode recording of heterotopic neurons in humans. Highly consistent interictal patterns (IPs) were identified within the nodules: 1) Periodic Discharges PLUS Fast activity (PD+F), Sporadic discharges PLUS Fast activity (SD+F), and 3) epileptic spikes (ES). Neuronal firing rates were significantly modulated during all IPs, suggesting that multiple IPs were generated by the same local neuronal populations. Furthermore, firing rates closely followed IP morphologies. Among the different IPs, SD+FA pattern was found only in the three nodules that were actively involved in seizure generation, but was never observed in the nodule that did not take part in ictal discharges. On the contrary, PD+F and ES were identified in all nodules. Units that were modulated during the IPs were also found to participate in seizures, increasing their firing rate at seizure onset and maintaining an elevated rate during the seizures. Together, nodules in PNH are highly epileptogenic, and show several IPs that provide promising pathognomonic signatures of PNH. Furthermore, our results show that PNH nodules may well initiate seizures. HighlightsO_LIFirst in vivo microelectrode description of local epileptic activities in human PNH C_LIO_LIRecordings revealed multiple microscopic epileptic interictal patterns C_LIO_LIFiring rates of all detected units were significantly modulated during all interictal patterns C_LIO_LISeizures recruited the same units that are involved in interictal activity C_LI

pathology↗