Dendritic effects of tDCS insights from a morphologically realistic model neuron.

Transcranial direct current stimulation (tDCS) induces subcellular compartmental-dependent polarization, maximal in the distal portions of axons and dendrites. Using a morphologically realistic neuron model, we simulated tDCS-induced membrane polarization of the apical dendrite. Thus, we investigated the differential dendritic effects of anodal and cathodal tDCS on membrane potential polarization along the dendritic structure and its subsequent effects on dendritic membrane resistance, excitatory postsynaptic potential amplitude, backpropagating action potential amplitude, input/output relations, and long-term synaptic plasticity. We further showed that the effects of anodal and cathodal tDCS on the backpropagating action potential were asymmetric, and explained this asymmetry. Additionally, we showed that the effects on input/output relations were rather weak and limited to the low-mid range of stimulation frequencies, and that synaptic plasticity effects were mostly limited to the distal portion of the dendrite. Thus, we demonstrated how tDCS modifies dendritic physiology due to the dendrite's unique morphology and composition of voltage-gated ion channels.

Molecular Insights into Transcranial Direct Current Stimulation Effects: Metabolomics and Transcriptomics Analyses.

INTRODUCTION: Transcranial direct current stimulation (tDCS) is an evolving non-invasive neurostimulation technique. Despite multiple studies, its underlying molecular mechanisms are still unclear. Several previous human studies of the effect of tDCS suggest that it generates metabolic effects. The induction of metabolic effects by tDCS could provide an explanation for how it generates its long-term beneficial clinical outcome. AIM: Given these hints of tDCS metabolic effects, we aimed to delineate the metabolic pathways involved in its mode of action. METHODS: To accomplish this, we utilized a broad analytical approach of co-analyzing metabolomics and transcriptomic data generated from anodal tDCS in rat models. Since no metabolomic dataset was available, we performed a tDCS experiment of bilateral anodal stimulation of 200 µA for 20 min and for 5 consecutive days, followed by harvesting the brain tissue below the stimulating electrode and generating a metabolomics dataset using LC-MS/MS. The analysis of the transcriptomic dataset was based on a publicly available dataset. RESULTS: Our analyses revealed that tDCS alters the metabolic profile of brain tissue, affecting bioenergetic-related pathways, such as glycolysis and mitochondrial functioning. In addition, we found changes in calcium-related signaling. CONCLUSIONS: We conclude that tDCS affects metabolism by modulating energy production-related processes. Given our findings concerning calcium-related signaling, we suggest that the immediate effects of tDCS on calcium dynamics drive modifications in distinct metabolic pathways. A thorough understanding of the underlying molecular mechanisms of tDCS has the potential to revolutionize its applicability, enabling the generation of personalized medicine in the field of neurostimulation and thus contributing to its optimization.

Direct Current Stimulation Modulates Synaptic Facilitation via Distinct Presynaptic Calcium Channels.

Transcranial direct current stimulation (tDCS) is a subthreshold neurostimulation technique known for ameliorating neuropsychiatric conditions. The principal mechanism of tDCS is the differential polarization of subcellular neuronal compartments, particularly the axon terminals that are sensitive to external electrical fields. Yet, the underlying mechanism of tDCS is not fully clear. Here, we hypothesized that direct current stimulation (DCS)-induced modulation of presynaptic calcium channel conductance alters axon terminal dynamics with regard to synaptic vesicle release. To examine the involvement of calcium-channel subtypes in tDCS, we recorded spontaneous excitatory postsynaptic currents (sEPSCs) from cortical layer-V pyramidal neurons under DCS while selectively inhibiting distinct subtypes of voltage-dependent calcium channels. Blocking P/Q or N-type calcium channels occluded the effects of DCS on sEPSCs, demonstrating their critical role in the process of DCS-induced modulation of spontaneous vesicle release. However, inhibiting T-type calcium channels did not occlude DCS-induced modulation of sEPSCs, suggesting that despite being active in the subthreshold range, T-type calcium channels are not involved in the axonal effects of DCS. DCS modulates synaptic facilitation by regulating calcium channels in axon terminals, primarily via controlling P/Q and N-type calcium channels, while T-type calcium channels are not involved in this mechanism.

Attitudes of psychiatrists toward telepsychiatry: A policy Delphi study.

Objectives: To delineate areas of consensus and disagreements among practicing psychiatrists from various levels of clinical experience, hierarchy and organizations, and to test their ability to converge toward agreement, which will enable better integration of telepsychiatry into mental health services. Methods: To study attitudes of Israeli public health psychiatrists, we utilized a policy Delphi method, during the early stages of the COVID pandemic. In-depth interviews were conducted and analyzed, and a questionnaire was generated. The questionnaire was disseminated amongst 49 psychiatrists, in two succeeding rounds, and areas of consensus and controversies were identified. Results: Psychiatrists showed an overall consensus regarding issues of economic and temporal advantages of telepsychiatry. However, the quality of diagnosis and treatment and the prospect of expanding the usage of telepsychiatry to normal circumstances-beyond situations of pandemic or emergency were disputed. Nonetheless, efficiency and willingness scales slightly improved during the 2nd round of the Delphi process. Prior experience with telepsychiatry had a strong impact on the attitude of psychiatrists, and those who were familiar with this practice were more favorable toward its usage in their clinic. Conclusions: We have delineated experience as a major impact on the attitudes toward telepsychiatry and the willingness for its assimilation in clinical practice as a legitimate and trustworthy method. We have also observed that the organizational affiliation significantly affected psychiatrists' attitude, when those working at local clinics were more positive toward telepsychiatry compared with employees of governmental institutions. This might be related to experience and differences in organizational environment. Taken together, we recommend to include hands-on training of telepsychiatry in medical education curriculum during residency, as well as refresher exercises for attending practitioners.

Synergies between synaptic and HCN channel plasticity dictates firing rate homeostasis and mutual information transfer in hippocampal model neuron.

Homeostasis is a precondition for any physiological system of any living organism. Nonetheless, models of learning and memory that are based on processes of synaptic plasticity are unstable by nature according to Hebbian rules, and it is not fully clear how homeostasis is maintained during these processes. This is where theoretical and computational frameworks can help in gaining a deeper understanding of the various cellular processes that enable homeostasis in the face of plasticity. A previous simplistic single compartmental model with a single synapse showed that maintaining input/output response homeostasis and stable synaptic learning could be enabled by introducing a linear relationship between synaptic plasticity and HCN conductance plasticity. In this study, we aimed to examine whether this approach could be extended to a more morphologically realistic model that entails multiple synapses and gradients of various VGICs. In doing so, we found that a linear relationship between synaptic plasticity and HCN conductance plasticity was able to maintain input/output response homeostasis in our morphologically realistic model, where the slope of the linear relationship was dependent on baseline HCN conductance and synaptic permeability values. An increase in either baseline HCN conductance or synaptic permeability value led to a decrease in the slope of the linear relationship. We further show that in striking contrast to the single compartment model, here linear relationship was insufficient in maintaining stable synaptic learning despite maintaining input/output response homeostasis. Additionally, we showed that homeostasis of input/output response profiles was at the expense of decreasing the mutual information transfer due to the increase in noise entropy, which could not be fully rescued by optimizing the linear relationship between synaptic and HCN conductance plasticity. Finally, we generated a place cell model based on theta oscillations and show that synaptic plasticity disrupts place cell activity. Whereas synaptic plasticity accompanied by HCN conductance plasticity through linear relationship maintains the stability of place cell activity. Our study establishes potential differences between a single compartmental model and a morphologically realistic model.

Elevated ROS levels during the early development of Angelman syndrome alter the apoptotic capacity of the developing neural precursor cells.

Angelman syndrome (AS) is a rare genetic neurodevelopmental disorder caused by the maternally inherited loss of function of the UBE3A gene. AS is characterized by a developmental delay, lack of speech, motor dysfunction, epilepsy, autistic features, happy demeanor, and intellectual disability. While the cellular roles of UBE3A are not fully understood, studies suggest that the lack of UBE3A function is associated with elevated levels of reactive oxygen species (ROS). Despite the accumulating evidence emphasizing the importance of ROS during early brain development and its involvement in different neurodevelopmental disorders, up to date, the levels of ROS in AS neural precursor cells (NPCs) and the consequences on AS embryonic neural development have not been elucidated. In this study we show multifaceted mitochondrial aberration in AS brain-derived embryonic NPCs, which exhibit elevated mitochondrial membrane potential (ΔΨm), lower levels of endogenous reduced glutathione, excessive mitochondrial ROS (mROS) levels, and increased apoptosis compared to wild-type (WT) littermates. In addition, we report that glutathione replenishment by glutathione-reduced ethyl ester (GSH-EE) corrects the excessive mROS levels and attenuates the enhanced apoptosis in AS NPCs. Studying the glutathione redox imbalance and mitochondrial abnormalities in embryonic AS NPCs provides an essential insight into the involvement of UBE3A in early neural development, information that can serve as a powerful avenue towards a broader view of AS pathogenesis. Moreover, since mitochondrial dysfunction and elevated ROS levels were associated with other neurodevelopmental disorders, the findings herein suggest some potential shared underlying mechanisms for these disorders as well.

Voltage-Gated Ion Channels and the Variability in Information Transfer.

The prerequisites for neurons to function within a circuit and be able to contain and transfer information efficiently and reliably are that they need to be homeostatically stable and fire within a reasonable range, characteristics that are governed, among others, by voltage-gated ion channels (VGICs). Nonetheless, neurons entail large variability in the expression levels of VGICs and their corresponding intrinsic properties, but the role of this variability in information transfer is not fully known. In this study, we aimed to investigate how this variability of VGICs affects information transfer. For this, we used a previously derived population of neuronal model neurons, each with the variable expression of five types of VGICs, fast Na+, delayed rectifier K+, A-type K+, T-type Ca++, and HCN channels. These analyses showed that the model neurons displayed variability in mutual information transfer, measured as the capability of neurons to successfully encode incoming synaptic information in output firing frequencies. Likewise, variability in the expression of VGICs caused variability in EPSPs and IPSPs amplitudes, reflected in the variability of output firing frequencies. Finally, using the virtual knockout methodology, we show that among the ion channels tested, the A-type K+ channel is the major regulator of information processing and transfer.

An Association Study of DNA Methylation and Gene Expression in Angelman Syndrome: A Bioinformatics Approach.

Angelman syndrome (AS) is a neurodevelopmental disorder caused by the loss of function of the E3-ligase UBE3A. Despite multiple studies, AS pathophysiology is still obscure and has mostly been explored in rodent models of the disease. In recent years, a growing body of studies has utilized omics datasets in the attempt to focus research regarding the pathophysiology of AS. Here, for the first time, we utilized a multi-omics approach at the epigenomic level and the transcriptome level, for human-derived neurons. Using publicly available datasets for DNA methylation and gene expression, we found genome regions in proximity to gene promoters and intersecting with gene-body regions that were differentially methylated and differentially expressed in AS. We found that overall, the genome in AS postmortem brain tissue was hypo-methylated compared to healthy controls. We also found more upregulated genes than downregulated genes in AS. Many of these dysregulated genes in neurons obtained from AS patients are known to be critical for neuronal development and synaptic functioning. Taken together, our results suggest a list of dysregulated genes that may be involved in AS development and its pathological features. Moreover, these genes might also have a role in neurodevelopmental disorders similar to AS.

The role of axonal voltage-gated potassium channels in tDCS.

BACKGROUND: Transcranial direct current stimulation (tDCS) is a non-invasive sub-threshold stimulation, widely accepted for its amelioration of distinct neuropsychiatric disorders. The weak electric field of tDCS modulates the activity of cortical neurons, which in turn modifies brain functioning. However, the underlying mechanisms for that are not fully understood. OBJECTIVE/HYPOTHESIS: Previous studies demonstrated that the axons are the most sensitive subcellular compartment for tDCS-induced polarization. Moreover, it was posited that DCS-induced axonal polarization is amplified by modifying the conductance of ionic channels. We posit that voltage-gated potassium-channels that are highly expressed in axons play a crucial role in DCS-induced modulation of cortical neurons functioning. METHODS: We examined the involvement of voltage-gated potassium-channels in the active modulation of spontaneous vesicle release by DCS. For that, we measured spontaneous excitatory postsynaptic currents (sEPSCs) from layer-V motor cortex during DCS application, while co-applying distinct voltage-gated potassium-channels blockers. Moreover, we examined the role of Kv1 potassium channels in DCS-induced modulation of action potential waveform at axon terminals by recording action potentials at terminal axon blebs during DCS application while locally inhibiting the Kv1 potassium-channels. RESULTS: We demonstrated that inhibiting voltage-gated potassium-channels occluded the DCS-induced modulation of subthreshold presynaptic vesicle release. Moreover, we showed that inhibiting Kv1 voltage-gated potassium-channels also occluded the DCS-induced modulation of action potential waveform at axon terminals. CONCLUSION: We suggest that DCS-induced depolarization inactivates the Kv1 potassium channels thus reducing potassium conductance, which amplifies axonal depolarization, subsequently enhancing the presynaptic component of synaptic transmission. Whereas DCS-induced hyperpolarization induces opposite effects.

CTCF in parvalbumin-expressing neurons regulates motor, anxiety and social behavior and neuronal identity.

CCCTC-binding factor (CTCF) is a regulator of chromatin organization and has direct effects on gene transcription. Mutations in CTCF have been identified in individuals with neurodevelopmental conditions. There are wide range of behaviors associated with these mutations, including intellectual disabilities, changes in temperament, and autism. Previous mice-model studies have identified roles for CTCF in excitatory neurons in specific behaviors, particularly in regards to learning and memory. However, the role of CTCF in inhibitory neurons is less well defined. In the current study, specific knockout of CTCF in parvalbumin-expressing neurons, a subset of inhibitory neurons, induced a specific behavioral phenotype, including locomotor abnormalities, anxiolytic behavior, and a decrease in social behavior. The anxiolytic and social abnormalities are detected before the onset of locomotor abnormalities. Immunohistochemical analysis revealed a disbalance in parvalbumin-expressing and somatostatin-expressing cells in these mice. Single nuclei RNA sequencing identified changes in gene expression in parvalbumin-expressing neurons that are specific to inhibitory neuronal identity and function. Electrophysiology analysis revealed an enhanced inhibitory tone in the hippocampal pyramidal neurons in knockout mice. These findings indicate that CTCF in parvalbumin-expressing neurons has a significant role in the overall phenotype of CTCF-associated neurodevelopmental deficits.

The Attitudes and Perceptions of Israeli Psychiatrists Toward Telepsychiatry and Their Behavioral Intention to Use Telepsychiatry.

Background: Although telemedicine care has grown in recent years, telepsychiatry is growing at a slower pace than expected, because service providers often hamper the assimilation and expansion of telepsychiatry due their attitudes and perceptions. The unified theory of acceptance and use of technology (UTAUT) is a model that was developed to assess the factors influencing the assimilation of a new technology. We used the UTAUT model to examine the associations between the attitudes and perceptions of psychiatrists in Israel toward telepsychiatry and their intention to use it. Methods: An online, close-ended questionnaire based on a modified UTAUT model was distributed among psychiatrists in Israel. Seventy-six questionnaires were completed and statistically analyzed. Results: The behavioral intention of Israeli psychiatrists to use telepsychiatry was relatively low, despite their perceptions of themselves as capable of high performance with low effort. Nonetheless, they were interested in using telepsychiatry voluntarily. Experience in telepsychiatry, and to a lesser extent, facilitating conditions, were found to be positively correlated with the intention to use telepsychiatry. Psychiatrists have a positive attitude toward treating patients by telepsychiatry and perceive its risk as moderate. Discussion: Despite high performance expectancy, low effort expectancy, low perceived risk, largely positive attitudes, high voluntariness, and the expectancy for facilitating conditions, the intention to use telepsychiatry was rather low. This result is explained by the low level of experience, which plays a pivotal role. We recommend promoting the facilitating conditions that affect the continued use of telepsychiatry when initiating its implementation, and conclude that it is critical to create a sense of success during the initial stages of experience.

Calcium channels control tDCS-induced spontaneous vesicle release from axon terminals.

BACKGROUND: Transcranial direct current stimulation (tDCS) is a subthreshold neurostimulation therapeutic method that ameliorate neuropsychiatric impairments. The most sensitive subcellular compartment for tDCS are the axons that polarize. However, how these relatively small polarizations significantly alter synaptic dynamics is still unknown. OBJECTIVE/HYPOTHESIS: We hypothesized that tDCS-induced axonal polarization modulates calcium channel activity at the presynaptic compartment, thus playing a crucial role in synaptic vesicle release. METHODS: For this aim, we examined how different DCS conditions and orientations affect the spontaneous excitatory post synaptic currents (sEPSCs) recorded from hippocampal CA1 pyramidal neurons. Since P/Q-type calcium-channels are the main presynaptic voltage-dependent calcium-channels in the hippocampus, we further examined the DCS effects while applying a P/Q-type calcium channels blocker, ω-agatoxin. Additionally, to explain the DCS-induced calcium channel-regulated vesicle release dynamics, we developed a simplified model to complement our experimental results. RESULTS: We demonstrated that anodal-DCS application in a dorso-ventral orientation, similar to that of in-vivo experiments, enhanced the sEPSCs frequency, while cathodal-DCS was ineffective. Moreover, DCS application in parallel to the Schaffer collaterals (medio-lateral orientation), showed both anodal and cathodal significant effects. Furthermore, the ω-agatoxin application occluded the DCS-induced modulation of sEPSC frequencies in any orientation. The model showed the interaction between DCS-induced membrane polarization, calcium channel activation and presynaptic vesicle release. CONCLUSION: Using experiments and modeling we show that DCS induces a small variation in terminal membrane potential sufficient to activate P/Q type voltage-gated calcium channels, and that this is sufficient to modify presynaptic calcium concentration, subsequently altering spontaneous vesicle release.

Attitudinal Barriers Hindering Adoption of Telepsychiatry among Mental Healthcare Professionals: Israel as a Case-Study.

Despite proven advantages for the use of telemedicine in psychiatry, mental healthcare professionals have shown deep-seated mistrust and suspicion of telepsychiatry, which hinders its widespread application. The current study examines the attitudes of Israeli mental health professionals towards telepsychiatry and seeks to uncover the effects of experience and organizational affiliation on its adoption. The methodology included qualitative and thematic analysis of 27 in-depth interviews with Israeli mental health professionals, focusing on three major themes-clinical quality, economic efficiency, and the effects on the work-life balance of healthcare professionals. The attitudes of mental health professionals were found to be widely divergent and sharply dichotomized regarding different aspects of telepsychiatry and its suitability for mental healthcare services. However, there was a general consensus that telemedicine may not fulfil its promise of being a panacea to the problems of modern public medicine. In addition, attitudes were related to hierarchical position, organizational affiliation, and personal experience with telepsychiatry. Specifically, organizational affiliation influenced experience with and support for the assimilation of telepsychiatry. The study also revealed the role of organizational leadership and culture in promoting or inhibiting the proliferation and adoption of innovative technologies and services in modern medicine.

The role of sodium channels in direct current stimulation-axonal perspective.

Transcranial neurostimulation methods are utilized as therapies for various neuropsychiatric disorders. Primarily, they entail the delivery of weak subthreshold currents across the brain, which modulate neuronal excitability. However, it is still a puzzle how such weak electrical fields actuate their effects. Previous studies showed that axons are the most sensitive subcellular compartment for direct current stimulation, and maximal polarization is achieved at their terminals. Nonetheless, polarization of axon terminals according to models was predicted to be weak, and the mechanism for substantial axon terminals polarization was obscure. Here, we show that a weak subthreshold electrical field modifies the conductance of voltage-dependent sodium channels in axon terminals, subsequently amplifying their membrane polarization. Moreover, we show that this amplification has substantial effects on synaptic functioning. Finally, we employ analytical modeling to explain how sodium currents modifications enhance axon terminal polarization. These findings relate to the mechanistic aspects of any neurostimulation technique.

Electrophysiological Characterization of Regular and Burst Firing Pyramidal Neurons of the Dorsal Subiculum in an Angelman Syndrome Mouse Model.

Angelman syndrome (AS) is a debilitating neurogenetic disorder characterized by severe developmental delay, speech impairment, gait ataxia, sleep disturbances, epilepsy, and a unique behavioral phenotype. AS is caused by a microdeletion or mutation in the maternal 15q11-q13 chromosome region containing UBE3A gene. The hippocampus is one of the important brain regions affected in AS mice leading to substantial hippocampal-dependent cognitive and behavioral deficits. Recent studies have suggested an abnormal increase in the α1-Na/K-ATPase (α1-NaKA) in AS mice as the precipitating factor leading to the hippocampal deficits. A subsequent study showed that the hippocampal-dependent behavioral deficits occur as a result of altered calcium (Ca+2) dynamics in the CA1 pyramidal neurons (PNs) caused by the elevated α1-NaKA expression levels in the AS mice. Nonetheless, a causal link between hippocampal deficits and major behavioral phenotypes in AS is still obscure. Subiculum, a region adjacent to the hippocampal CA1 is the major output source of the hippocampus and plays an important role in the transfer of information from the CA1 region to the cortical areas. However, in spite of the robust hippocampal deficits and several known electrophysiological alterations in multiple brain regions in AS mice, the neuronal properties of the subicular neurons were never investigated in these mice. Additionally, subicular function is also implied in many neuropsychiatric disorders such as autism, schizophrenia, Alzheimer's disease, and epilepsy that share some common features with AS. Therefore, given the importance of the subiculum in these neuropsychiatric disorders and the altered electrophysiological properties of the hippocampal CA1 PNs projecting to the subiculum, we sought to examine the subicular PNs. We performed whole-cell recordings from dorsal subiculum of both WT and AS mice and found three distinct populations of PNs based on their ability to fire bursts or single action potentials following somatic current injection: strong bursting, weak bursting, and regular firing neurons. We found no overall differences in the distribution of these different subicular PN populations among AS and WT controls. However, the different cell types showed distinct alterations in their intrinsic membrane properties. Further, none of these populations were altered in their excitatory synaptic properties. Altogether, our study characterized the different subtypes of PNs in the subicular region of an AS mouse model.

Angelman Syndrome and Angelman-like Syndromes Share the Same Calcium-Related Gene Signatures.

Angelman-like syndromes are a group of neurodevelopmental disorders that entail clinical presentation similar to Angelman Syndrome (AS). In our previous study, we showed that calcium signaling is disrupted in AS, and we identified calcium-target and calcium-regulating gene signatures that are able to differentiate between AS and their controls in different models. In the herein study, we evaluated these sets of calcium-target and calcium-regulating genes as signatures of AS-like and non-AS-like syndromes. We collected a number of RNA-seq datasets of various AS-like and non-AS-like syndromes and performed Principle Component Analysis (PCA) separately on the two sets of signature genes to visualize the distribution of samples on the PC1-PC2 plane. In addition to the evaluation of calcium signature genes, we performed differential gene expression analyses to identify calcium-related genes dysregulated in each of the studied syndromes. These analyses showed that the calcium-target and calcium-regulating signatures differentiate well between AS-like syndromes and their controls. However, in spite of the fact that many of the non-AS-like syndromes have multiple differentially expressed calcium-related genes, the calcium signatures were not efficient classifiers for non-AS-like neurodevelopmental disorders. These results show that features based on clinical presentation are reflected in signatures derived from bioinformatics analyses and suggest the use of bioinformatics as a tool for classification.

Aberrant aggressive behavior in a mouse model of Angelman syndrome.

Angelman syndrome (AS) is a genetic neurodevelopmental disorder due to the absence of the E3-ligase protein, UBE3A. Inappropriate social interactions, usually hyper-sociability, is a part of that syndrome. In addition, clinical surveys and case reports describe aggressive behavior in AS individuals as a severe difficulty for caretakers. A mouse model for AS recapitulates most of the human AS phenotypes. However, very few studies utilized this mouse model for investigating affiliative social behavior, and not even a single study examined aggressive behavior. Hence, the aim of the herein study was to examine affiliative and aggressive social behavior. For that, we utilized a battery of behavioral paradigms, and performed detailed analyses of these behaviors. AS mice exhibited a unique characteristic of reduced habituation towards a social stimulus in comparison to their wild-type (WT) littermates. However, overall there were no additional marked differences in affiliative social behavior. In contrast to the mild changes in affiliative behavior, there was a striking enhanced aggression in the AS mice compared to their WT littermates. The herein findings emphasize the use of AS mouse model in characterizing and measuring inappropriate aggressive behavior, and suggests these as tools for investigating therapeutic interventions aimed at attenuating aggressive behavior.

Bioinformatics analyses show dysregulation of calcium-related genes in Angelman syndrome mouse model.

BACKGROUND: Angelman syndrome (AS) is a genetic neurodevelopmental disorder caused by the loss of function of the UBE3A protein in the brain. In a previous study, we showed that activity-dependent calcium dynamics in hippocampal CA1 pyramidal neurons of AS mice is compromised, and its normalization rescues the hippocampal-dependent deficits. Therefore, we expected that the expression profiles of calcium-related genes would be altered in AS mice hippocampi. METHODS: We analyzed mRNA sequencing data from AS model mice and WT controls in light of the newly published CaGeDB database of calcium-related genes. We validated our results in two independent RNA sequencing datasets from two additional different AS models: first one, a human neuroblastoma cell line where UBE3A expression was knocked down by siRNA, and the second, an iPSC-derived neurons from AS patient and healthy donor control. FINDINGS: We found signatures of dysregulated calcium-related genes in AS mouse model hippocampus. Additionally, we show that these calcium-related genes function as signatures for AS in other human cellular models of AS, thus strengthening our findings. INTERPRETATION: Our findings suggest the downstream implications and significance of the compromised calcium signaling in Angelman syndrome. Moreover, since AS share similar features with other autism spectrum disorders, we believe that these findings entail meaningful data and approach for other neurodevelopmental disorders, especially those with known alterations of calcium signaling. FUNDING: This work was supported by the Angelman Syndrome Foundation and by the Israel Science Foundation, Grant Number 248/20.

Chronic α1-Na/K-ATPase inhibition reverses the elongation of the axon initial segment of the hippocampal CA1 pyramidal neurons in Angelman syndrome model mice.

Angelman syndrome (AS) is a neurodevelopmental disorder caused by the loss of function of the maternal UBE3A gene. The hippocampus is one of the most prominently affected brain regions in AS model mice, manifesting in severe hippocampal-dependent memory and plasticity deficits. Previous studies in AS mice reported an elongated axon initial segment (AIS) in pyramidal neurons (PNs) of the hippocampal CA1 region. These were the first reports in mammals to show AIS elongation in vivo. Correspondingly, this AIS elongation was linked to enhanced expression of the α1 subunit of Na+/K+-ATPase (α1-NaKA). Recently, it was shown that selective pharmacological inhibition of α1-NaKA by marinobufagenin (MBG) in adult AS mice rescued the hippocampal-dependent deficits via normalizing their compromised activity-dependent calcium (Ca+2) dynamics. In the herein study, we showed that a chronic selective α1-NaKA inhibition reversed the AIS elongation in hippocampal CA1 PNs of adult AS mice, and differentially altered their excitability and intrinsic properties. Taken together, our study is the first to demonstrate in vivo structural plasticity of the AIS in a mammalian model, and further elaborates on the modulatory effects of elevated α1-NaKA levels in the hippocampus of AS mice.

Bioinformatics Analyses of the Transcriptome Reveal Ube3a-Dependent Effects on Mitochondrial-Related Pathways.

The UBE3A gene encodes the ubiquitin E3-ligase protein, UBE3A, which is implicated in severe neurodevelopmental disorders. Lack of UBE3A expression results in Angelman syndrome, while UBE3A overexpression, due to genomic 15q duplication, results in autism. The cellular roles of UBE3A are not fully understood, yet a growing body of evidence indicates that these disorders involve mitochondrial dysfunction and increased oxidative stress. We utilized bioinformatics approaches to delineate the effects of murine Ube3a deletion on the expression of mitochondrial-related genes and pathways. For this, we generated an mRNA sequencing dataset from mouse embryonic fibroblasts (MEFs) in which both alleles of Ube3a gene were deleted and their wild-type controls. Since oxidative stress and mitochondrial dysregulation might not be exhibited in the resting baseline state, we also activated mitochondrial functioning in the cells of these two genotypes using TNFα application. Transcriptomes of the four groups of MEFs, Ube3a+/+ and Ube3a-/-, with or without the application of TNFα, were analyzed using various bioinformatics tools and machine learning approaches. Our results indicate that Ube3a deletion affects the gene expression profiles of mitochondrial-associated pathways. We further confirmed these results by analyzing other publicly available human transcriptome datasets of Angelman syndrome and 15q duplication syndrome.