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Multifractal analysis offers a sophisticated method to examine the complex morphology of neurons, which traditionally have been analyzed using monofractal techniques. This study investigates the multifractal properties of two-dimensional neuron projections from the human dorsal striatum, focusing on potential morphological changes related to aging and differences based on spatial origin within the nucleus. Using multifractal spectra, we analyzed various parameters, including generalized dimensions and Hölder exponents, to characterize the neurons' morphology. Despite the detailed analysis, no significant correlation was found between neuronal morphology and age. However, clear morphological differences were observed between neurons from the caudate nucleus and the putamen. Neurons from the putamen displayed higher morphological complexity and greater local homogeneity, while those from the caudate nucleus exhibited more scaling laws and higher local heterogeneity. These findings suggest that while age may not significantly impact neuronal morphology in the dorsal striatum, the spatial origin within this brain region plays a crucial role in determining neuronal structure. Further studies with larger samples are recommended to confirm these findings and to explore the full potential of multifractal analysis in neuronal morphology research. [ABSTRACT FROM AUTHOR]

Murine organotypic brain slice cultures have been widely used in neuroscientific research and are offering the opportunity to study neuronal function under normal and disease conditions. Despite the broad application, the mechanisms governing the maturation of immature cortical circuits in vitro are not well understood. In this study, we present a detailed investigation into the development of the neocortex in vitro. Using a holistic approach, we studied organotypic whole hemisphere brain slice cultures from postnatal mice and tracked the development of the somatosensory area over a 5-wk period. Our analysis revealed the maturation of passive and active intrinsic properties of pyramidal cells together with their morphology, closely resembling in vivo development. Detailed multielectrode array (MEA) electrophysiological assessments and RNA expression profiling demonstrated stable network properties by 2 wk in culture, followed by the transition of spontaneous activity toward more complex patterns including high-frequency oscillations. However, culturing weeks 4 and 5 exhibited increased variability and initial signs of neuronal loss, highlighting the importance of considering developmental stages in experimental design. This comprehensive characterization is vital for understanding the temporal dynamics of the neocortical development in vitro, with implications for neuroscientific research methodologies, particularly in the investigation of diseases such as epilepsy and other neurodevelopmental disorders. NEW & NOTEWORTHY: The development of the mouse neocortex in vitro mimics the in vivo development. Mouse brain cultures can serve as a model system for cortical development for the first 2 wk in vitro and as a model system for the adult cortex from 2 to 4 wk in vitro. Mouse organotypic brain slice cultures develop high-frequency network oscillations at γ frequency after 2 wk in vitro. Mouse brain cultures exhibit increased heterogeneity and variability after 4 wk in culture. [ABSTRACT FROM AUTHOR]

Background Dendritic spines are tiny protrusions found along the dendrites of neurons, and their number is a measure of the density of synaptic connections. Altered density and morphology is observed in several pathologies, and spine formation as well as morphological changes correlate with learning and memory. The detection of spines in microscopy images and the analysis of their morphology is therefore a prerequisite for many studies. We have developed a new open-source, freely available, plugin for ImageJ/FIJI, called Spot Spine, that allows detection and morphological measurements of spines in three dimensional images. Method Local maxima are detected in spine heads, and the intensity distribution around the local maximum is computed to perform the segmentation of each spine head. Spine necks are then traced from the spine head to the dendrite. Several parameters can be set to optimize detection and segmentation, and manual correction gives further control over the result of the process. Results The plugin allows the analysis of images of dendrites obtained with various labeling and imaging methods. Quantitative measurements are retrieved including spine head volume and surface, and neck length. Conclusion The plugin and instructions for use are available at https://imagej.net/plugins/spot-spine.

Golgi staining is a classical method for visualization of neuronal dendrites and dendritic spines and is still widely used for neuronal morphology visualization. Silver Golgi staining currently suffers from problems such as the tendency to produce black precipitation and incomplete morphological structure of stained neurons. We investigated how potassium dichromate (K2Cr2O7 and silver nitrate (AgNO3 affect the staining effect of the silver Golgi staining method, and realized the optimization of the silver Golgi staining method by studying the change of the mass concentration of the staining solution constituents, potassium dichromate, and silver nitrate, and the different modes of action of the components. The results showed that： the staining effect of the mixture of 35 mg/mL paraformaldehyde (PFA and 20 mg/mL K2Cr2O7 was significantly better than that of 20 mg/mL K2Cr2O7 alone, and during the optimization process of the mass concentration of K2Cr2O7, the best staining effect was achieved by 80 mg/mL K2Cr2O7, and the staining effect of 35 mg /mL PFA mixed with 20 mg/mL K2Cr2O7 and 80 mg/mL K2Cr2O7 optimal staining duration ratio was 3：1, the optimal mass concentration of AgNO3 during the staining process was 10 mg/mL, and in the final staining solution composition and the mode of action of 2 days 40 mg/mL PFA fixation, 3 days 35 mg/mL PFA and 20 mg/mL K2Cr2O7, 40 mg/mL PFA fixation, 3 days 35 mg/mL PFA and 20 mg/mL K2Cr2O7, and 2 days 35 mg/mL PFA and 20 mg/mL K2Cr2O7. PFA and 20 mg/mL K2Cr2O7 mixed solution for 3 days, 80 mg/mL K2Cr2O7 for 1 day and 10 mg/mL AgNO3 for 5 days had the best effect on the visualization of neuronal morphology in brain tissue. The optimized silver-stained Golgi staining method will achieve complete morphological structural staining of neurons, and will also produce less imaging impurity precipitation noise during the staining process. This work will provide a new modified Golgi staining method to achieve staining of the complete morphological structure of neurons. This optimized method is expected to provide a new staining method for studying the relationship between structural changes of brain neurons and brain diseases.. [ABSTRACT FROM AUTHOR]

Neuronal morphology can be represented using various feature representations, such as hand-crafted morphometrics and deep features. These features are complementary to each other, contributing to improving performance. However, existing classification methods only utilize a single feature representation or simply concatenate different features without fully considering their complementarity. Therefore, their performance is limited and can be further improved. In this paper, we propose a multi-level feature fusion network that fully utilizes diverse feature representations and their complementarity to effectively describe neuronal morphology and improve performance. Specifically, we devise a Multi-Level Fusion Module (MLFM) and incorporate it into each feature extraction block. It can facilitate the interaction between different features and achieve effective feature fusion at multiple levels. The MLFM comprises a channel attention-based Feature Enhancement Module (FEM) and a cross-attention-based Feature Interaction Module (FIM). The FEM is used to enhance robust morphological feature presentations, while the FIM mines and propagates complementary information across different feature presentations. In this way, our feature fusion network ultimately yields a more distinctive neuronal morphology descriptor that can effectively characterize neurons than any singular morphological representation. Experimental results show that our method effectively depicts neuronal morphology and correctly classifies 10-type neurons on the NeuronMorpho-10 dataset with an accuracy of 95.18%, outperforming other approaches. Moreover, our method performs well on the NeuronMorpho-12 and NeuronMorpho-17 datasets and possesses good generalization.

Background Dendritic spines are tiny protrusions found along the dendrites of neurons, and their number is a measure of the density of synaptic connections. Altered density and morphology is observed in several pathologies, and spine formation as well as morphological changes correlate with learning and memory. The detection of spines in microscopy images and the analysis of their morphology is therefore a prerequisite for many studies. We have developed a new open-source, freely available, plugin for ImageJ/FIJI, called Spot Spine, that allows detection and morphological measurements of spines in three dimensional images. Method Local maxima are detected in spine heads, and the intensity distribution around the local maximum is computed to perform the segmentation of each spine head. Spine necks are then traced from the spine head to the dendrite. Several parameters can be set to optimize detection and segmentation, and manual correction gives further control over the result of the process. Results The plugin allows the analysis of images of dendrites obtained with various labeling and imaging methods. Quantitative measurements are retrieved including spine head volume and surface, and neck length. Conclusion The plugin and instructions for use are available at https://imagej.net/plugins/spot-spine.

Gulf War Illness (GWI) is a multi-symptom disorder that manifests with fatigue, sleep disturbances, mood-cognition pathologies, and musculoskeletal symptoms. GWI affects at least 25% of the military personnel that served in Operations Desert Shield and Desert Storm from 1990 to 1991. We modeled Gulf War toxicant exposure in C57BL/6J mice by combined exposure to pyridostigmine bromide (an anti-sarin drug), chlorpyrifos (an organophosphate insecticide), and DEET (an insect repellent) for 10 days followed by oral treatment with Withania somnifera root extract for 21 days beginning at 12 weeks post-exposure. W. somnifera, commonly referred to as ashwagandha, has been used in traditional Ayurvedic medicine for centuries to improve memory and reduce inflammation, and its roots contain bioactive molecules which share functional groups with modern pain, cancer, and anti-inflammatory drugs. Previously, we observed that GWI mice displayed chronic reductions in dendritic arbor and loss of spines in granule cells of the dentate gyrus of the hippocampus at 14 weeks post-exposure. Here, we examined the effects of treatment with W. somnifera root extract on chronic dendrite and spine morphology in dentate granule cells of the mouse hippocampus following Gulf War toxicant exposure. GWI mice showed approximately 25% decreases in dendritic length (p < 0.0001) and overall dendritic spine density with significant reductions in thin and mushroom spines. GWI mice treated with the Ayurvedic W. somnifera extract exhibited dendritic lengths and spine densities near normal levels. These findings demonstrate the efficacy of the Ayurvedic treatment for neuroprotection following these toxic exposures. We hope that the extract and the neuronal processes influenced will open new avenues of research regarding treatment of Gulf War Illness and neurodegenerative disorders. [ABSTRACT FROM AUTHOR]

Neurons typically generate action potentials at their axon initial segment based on the integration of synaptic inputs. In many neurons, the axon extends from the soma, equally weighting dendritic inputs. A notable exception is found in a subset of hippocampal pyramidal cells where the axon emerges from a basal dendrite. This structure allows these axon-carrying dendrites (AcDs) a privileged input route. We found that in male mice, such cells in the CA1 region receive stronger excitatory input from the contralateral CA3, compared with those with somatic axon origins. This is supported by a higher count of putative synapses from contralateral CA3 on the AcD. These findings, combined with prior observations of their distinct role in sharp-wave ripple firing, suggest a key role of this neuron subset in coordinating bi-hemispheric hippocampal activity during memory-centric oscillations. [ABSTRACT FROM AUTHOR]

Aging is the leading risk factor for Alzheimer's disease and other neurodegenerative diseases. We now understand that a breakdown in the neuronal cytoskeleton, mainly underpinned by protein modifications leading to the destabilization of microtubules, is central to the pathogenesis of Alzheimer's disease. This is accompanied by morphological defects across the somatodendritic compartment, axon, and synapse. However, knowledge of what occurs to the microtubule cytoskeleton and morphology of the neuron during physiological aging is comparatively poor. Several recent studies have suggested that there is an age-related increase in the phosphorylation of the key microtubule stabilizing protein tau, a modification, which is known to destabilize the cytoskeleton in Alzheimer's disease. This indicates that the cytoskeleton and potentially other neuronal structures reliant on the cytoskeleton become functionally compromised during normal physiological aging. The current literature shows age-related reductions in synaptic spine density and shifts in synaptic spine conformation which might explain age-related synaptic functional deficits. However, knowledge of what occurs to the microtubular and actin cytoskeleton, with increasing age is extremely limited. When considering the somatodendritic compartment, a regression in dendrites and loss of dendritic length and volume is reported whilst a reduction in soma volume/size is often seen. However, research into cytoskeletal change is limited to a handful of studies demonstrating reductions in and mislocalizations of microtubule-associated proteins with just one study directly exploring the integrity of the microtubules. In the axon, an increase in axonal diameter and age-related appearance of swellings is reported but like the dendrites, just one study investigates the microtubules directly with others reporting loss or mislocalization of microtubule-associated proteins. Though these are the general trends reported, there are clear disparities between model organisms and brain regions that are worthy of further investigation. Additionally, longitudinal studies of neuronal/cytoskeletal aging should also investigate whether these age-related changes contribute not just to vulnerability to disease but also to the decline in nervous system function and behavioral output that all organisms experience. This will highlight the utility, if any, of cytoskeletal fortification for the promotion of healthy neuronal aging and potential protection against age-related neurodegenerative disease. This review seeks to summarize what is currently known about the physiological aging of the neuron and microtubular cytoskeleton in the hope of uncovering mechanisms underpinning age-related risk to disease.

Multifractal analysis offers a sophisticated method to examine the complex morphology of neurons, which traditionally have been analyzed using monofractal techniques. This study investigates the multifractal properties of two-dimensional neuron projections from the human dorsal striatum, focusing on potential morphological changes related to aging and differences based on spatial origin within the nucleus. Using multifractal spectra, we analyzed various parameters, including generalized dimensions and Hölder exponents, to characterize the neurons’ morphology. Despite the detailed analysis, no significant correlation was found between neuronal morphology and age. However, clear morphological differences were observed between neurons from the caudate nucleus and the putamen. Neurons from the putamen displayed higher morphological complexity and greater local homogeneity, while those from the caudate nucleus exhibited more scaling laws and higher local heterogeneity. These findings suggest that while age may not significantly impact neuronal morphology in the dorsal striatum, the spatial origin within this brain region plays a crucial role in determining neuronal structure. Further studies with larger samples are recommended to confirm these findings and to explore the full potential of multifractal analysis in neuronal morphology research.

Background Dendritic spines are tiny protrusions found along the dendrites of neurons, and their number is a measure of the density of synaptic connections. Altered density and morphology is observed in several pathologies, and spine formation as well as morphological changes correlate with learning and memory. The detection of spines in microscopy images and the analysis of their morphology is therefore a prerequisite for many studies. We have developed a new open-source, freely available, plugin for ImageJ/FIJI, called Spot Spine, that allows detection and morphological measurements of spines in three dimensional images. Method Local maxima are detected in spine heads, and the intensity distribution around the local maximum is computed to perform the segmentation of each spine head. Spine necks are then traced from the spine head to the dendrite. Several parameters can be set to optimize detection and segmentation, and manual correction gives further control over the result of the process. Results The plugin allows the analysis of images of dendrites obtained with various labeling and imaging methods. Quantitative measurements are retrieved including spine head volume and surface, and neck length. Conclusion The plugin and instructions for use are available at https://imagej.net/plugins/spot-spine.

Synapse formation is critical for the wiring of neural circuits in the developing brain. The synaptic scaffolding protein S-SCAM/MAGI-2 has important roles in the assembly of signaling complexes at post-synaptic densities. However, the role of S-SCAM in establishing the entire synapse is not known. Here, we report significant effects of RNAi-induced S-SCAM knockdown on the number of synapses in early stages of network development in vitro. In vivo knockdown during the first three postnatal weeks reduced the number of dendritic spines in the rat brain neocortex. Knockdown of S-SCAM in cultured hippocampal neurons severely reduced the clustering of both pre- and post-synaptic components. This included synaptic vesicle proteins, pre- and post-synaptic scaffolding proteins, and cell adhesion molecules, suggesting that entire synapses fail to form. Correspondingly, functional and morphological characteristics of developing neurons were affected by reducing S-SCAM protein levels; neurons displayed severely impaired synaptic transmission and reduced dendritic arborization. A next-generation sequencing approach showed normal expression of housekeeping genes but changes in expression levels in 39 synaptic signaling molecules in cultured neurons. These results indicate that S-SCAM mediates the recruitment of all key classes of synaptic molecules during synapse assembly and is critical for the development of neural circuits in the developing brain. [ABSTRACT FROM AUTHOR]

Gulf War Illness (GWI) is a multi-symptom disorder that manifests with fatigue, sleep disturbances, mood-cognition pathologies, and musculoskeletal symptoms. GWI affects at least 25% of the military personnel that served in Operations Desert Shield and Desert Storm from 1990 to 1991. We modeled Gulf War toxicant exposure in C57BL/6J mice by combined exposure to pyridostigmine bromide (an anti-sarin drug), chlorpyrifos (an organophosphate insecticide), and DEET (an insect repellent) for 10 days followed by oral treatment with Withania somnifera root extract for 21 days beginning at 12 weeks post-exposure. W. somnifera, commonly referred to as ashwagandha, has been used in traditional Ayurvedic medicine for centuries to improve memory and reduce inflammation, and its roots contain bioactive molecules which share functional groups with modern pain, cancer, and anti-inflammatory drugs. Previously, we observed that GWI mice displayed chronic reductions in dendritic arbor and loss of spines in granule cells of the dentate gyrus of the hippocampus at 14 weeks post-exposure. Here, we examined the effects of treatment with W. somnifera root extract on chronic dendrite and spine morphology in dentate granule cells of the mouse hippocampus following Gulf War toxicant exposure. GWI mice showed approximately 25% decreases in dendritic length (p < 0.0001) and overall dendritic spine density with significant reductions in thin and mushroom spines. GWI mice treated with the Ayurvedic W. somnifera extract exhibited dendritic lengths and spine densities near normal levels. These findings demonstrate the efficacy of the Ayurvedic treatment for neuroprotection following these toxic exposures. We hope that the extract and the neuronal processes influenced will open new avenues of research regarding treatment of Gulf War Illness and neurodegenerative disorders.

Synapse formation is critical for the wiring of neural circuits in the developing brain. The synaptic scaffolding protein S-SCAM/MAGI-2 has important roles in the assembly of signaling complexes at post-synaptic densities. However, the role of S-SCAM in establishing the entire synapse is not known. Here, we report significant effects of RNAi-induced S-SCAM knockdown on the number of synapses in early stages of network development in vitro. In vivo knockdown during the first three postnatal weeks reduced the number of dendritic spines in the rat brain neocortex. Knockdown of S-SCAM in cultured hippocampal neurons severely reduced the clustering of both pre- and post-synaptic components. This included synaptic vesicle proteins, pre- and post-synaptic scaffolding proteins, and cell adhesion molecules, suggesting that entire synapses fail to form. Correspondingly, functional and morphological characteristics of developing neurons were affected by reducing S-SCAM protein levels; neurons displayed severely impaired synaptic transmission and reduced dendritic arborization. A next-generation sequencing approach showed normal expression of housekeeping genes but changes in expression levels in 39 synaptic signaling molecules in cultured neurons. These results indicate that S-SCAM mediates the recruitment of all key classes of synaptic molecules during synapse assembly and is critical for the development of neural circuits in the developing brain.

Background: Spinocerebellar ataxia type 1 (SCA1) is a neurodegenerative disease caused by a polyglutamine expansion in the ataxin‐1 protein resulting in neuropathology including mutant ataxin‐1 protein aggregation, aberrant neurodevelopment, and mitochondrial dysfunction. Objectives: Identify SCA1‐relevant phenotypes in patient‐specific fibroblasts and SCA1 induced pluripotent stem cells (iPSCs) neuronal cultures. Methods: SCA1 iPSCs were generated and differentiated into neuronal cultures. Protein aggregation and neuronal morphology were evaluated using fluorescent microscopy. Mitochondrial respiration was measured using the Seahorse Analyzer. The multi‐electrode array (MEA) was used to identify network activity. Finally, gene expression changes were studied using RNA‐seq to identify disease‐specific mechanisms. Results: Bioenergetics deficits in patient‐derived fibroblasts and SCA1 neuronal cultures showed altered oxygen consumption rate, suggesting involvement of mitochondrial dysfunction in SCA1. In SCA1 hiPSC‐derived neuronal cells, nuclear and cytoplasmic aggregates were identified similar in localization as aggregates in SCA1 postmortem brain tissue. SCA1 hiPSC‐derived neuronal cells showed reduced dendrite length and number of branching points while MEA recordings identified delayed development in network activity in SCA1 hiPSC‐derived neuronal cells. Transcriptome analysis identified 1050 differentially expressed genes in SCA1 hiPSC‐derived neuronal cells associated with synapse organization and neuron projection guidance, where a subgroup of 151 genes was highly associated with SCA1 phenotypes and linked to SCA1 relevant signaling pathways. Conclusions: Patient‐derived cells recapitulate key pathological features of SCA1 pathogenesis providing a valuable tool for the identification of novel disease‐specific processes. This model can be used for high throughput screenings to identify compounds, which may prevent or rescue neurodegeneration in this devastating disease. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society. [ABSTRACT FROM AUTHOR]

Aims: Dynamin‐2 is a large GTPase, a member of the dynamin superfamily that regulates membrane remodelling and cytoskeleton dynamics. Mutations in the dynamin‐2 gene (DNM2) cause autosomal dominant centronuclear myopathy (CNM), a congenital neuromuscular disorder characterised by progressive weakness and atrophy of the skeletal muscles. Cognitive defects have been reported in some DNM2‐linked CNM patients suggesting that these mutations can also affect the central nervous system (CNS). Here we studied how a dynamin‐2 CNM‐causing mutation influences the CNS function. Methods: Heterozygous mice harbouring the p.R465W mutation in the dynamin–2 gene (HTZ), the most common causing autosomal dominant CNM, were used as disease model. We evaluated dendritic arborisation and spine density in hippocampal cultured neurons, analysed excitatory synaptic transmission by electrophysiological field recordings in hippocampal slices, and evaluated cognitive function by performing behavioural tests. Results: HTZ hippocampal neurons exhibited reduced dendritic arborisation and lower spine density than WT neurons, which was reversed by transfecting an interference RNA against the dynamin‐2 mutant allele. Additionally, HTZ mice showed defective hippocampal excitatory synaptic transmission and reduced recognition memory compared to the WT condition. Conclusion: Our findings suggest that the dynamin‐2 p.R465W mutation perturbs the synaptic and cognitive function in a CNM mouse model and support the idea that this GTPase plays a key role in regulating neuronal morphology and excitatory synaptic transmission in the hippocampus. [ABSTRACT FROM AUTHOR]

Single-cell transcriptomic approaches are revolutionizing neuroscience. Integrating this wealth of data with morphology and physiology, for the comprehensive study of neuronal biology, requires multiplexing gene expression data with complementary techniques. To meet this need, multiple groups in parallel have developed "Patch-seq," a modification of whole-cell patch-clamp protocols that enables mRNA sequencing of cell contents after electrophysiological recordings from individual neurons and morphologic reconstruction of the same cells. In this review, we first outline the critical technical developments that enabled robust Patch-seq experimental efforts and analytical solutions to interpret the rich multimodal data generated. We then review recent applications of Patch-seq that address novel and long-standing questions in neuroscience. These include the following: (1) targeted study of specific neuronal populations based on their anatomic location, functional properties, lineage, or a combination of these factors; (2) the compilation and integration of multimodal cell type atlases; and (3) the investigation of the molecular basis of morphologic and functional diversity. Finally, we highlight potential opportunities for further technical development and lines of research that may benefit from implementing the Patch-seq technique. As a multimodal approach at the intersection of molecular neurobiology and physiology, Patch-seq is uniquely positioned to directly link gene expression to brain function.

Duplications of the Xq28 region are a common cause of X‐linked intellectual disability (XLID). The RAB39B gene locates in Xq28 and has been implicated in disease pathogenesis. However, whether increased dosage of RAB39B leads to cognitive impairment and synaptic dysfunction remains elusive. Herein, we overexpressed RAB39B in mouse brain by injecting AAVs into bilateral ventricles of neonatal animals. We found that at 2 months of age, neuronal overexpression of RAB39B impaired the recognition memory and the short‐term working memory in mice and resulted in certain autism‐like behaviours, including social novelty defect and repetitive grooming behaviour in female mice. Moreover, overexpression of RAB39B decreased dendritic arborization of primary neurons in vitro and reduced synaptic transmission in female mice. Neuronal overexpression of RAB39B also altered autophagy without affecting levels and PSD distribution of synaptic proteins. Our results demonstrate that overexpression of RAB39B compromises normal neuronal development, thereby resulting in dysfunctional synaptic transmission and certain intellectual disability and behavioural abnormalities in mice. These findings identify a molecular mechanism underlying XLID with increased copy numbers of Xq28 and provide potential strategies for disease intervention. [ABSTRACT FROM AUTHOR]

Visualizing nerve cells has been fundamental for the systematic description of brain structure and function in humans and other species. Different approaches aimed to unravel the morphological features of neuron types and diversity. The inherent complexity of the human nervous tissue and the need for proper histological processing have made studying human dendrites and spines challenging in postmortem samples. In this study, we used Golgi data and open‐source software for 3D image reconstruction of human neurons from the cortical amygdaloid nucleus to show different dendrites and pleomorphic spines at different angles. Procedures required minimal equipment and generated high‐quality images for differently shaped cells. We used the "single‐section" Golgi method adapted for the human brain to engender 3D reconstructed images of the neuronal cell body and the dendritic ramification by adopting a neuronal tracing procedure. In addition, we elaborated 3D reconstructions to visualize heterogeneous dendritic spines using a supervised machine learning‐based algorithm for image segmentation. These tools provided an additional upgrade and enhanced visual display of information related to the spatial orientation of dendritic branches and for dendritic spines of varied sizes and shapes in these human subcortical neurons. This same approach can be adapted for other techniques, areas of the central or peripheral nervous system, and comparative analysis between species. [ABSTRACT FROM AUTHOR]

Parental care by fathers enhances offspring survival and development in numerous species. In the biparental California mouse, Peromyscus californicus, behavioral plasticity is seen during the transition into fatherhood: adult virgin males often exhibit aggressive or indifferent responses to pups, whereas fathers engage in extensive paternal care. In this species and other biparental mammals, the onset of paternal behavior is associated with increased neural responsiveness to pups in specific brain regions, including the medial preoptic area of the hypothalamus (MPOA), a region strongly implicated in both maternal and paternal behavior. To assess possible changes in neural circuit properties underlying this increased excitability, we evaluated synaptic, intrinsic, and morphological properties of MPOA neurons in adult male California mice that were either virgins or first-time fathers. We used standard whole-cell recordings in a novel in vitro slice preparation. Excitatory and inhibitory post-synaptic currents from MPOA neurons were recorded in response to local electrical stimulation, and input/output curves were constructed for each. Responses to trains of stimuli were also examined. We quantified intrinsic excitability by measuring voltage changes in response to square-pulse injections of both depolarizing and hyperpolarizing current. Biocytin was injected into neurons during recording, and their morphology was analyzed. Most parameters did not differ significantly between virgins and fathers. However, we document a decrease in synaptic inhibition in fathers. These findings suggest that the onset of paternal behavior in California mouse fathers may be associated with limited electrophysiological plasticity within the MPOA.

Here, we highlight the potential translational benefits of delivering FLASH radiotherapy using ultra-high dose rates (>100 Gy⋅s-1). Compared with conventional dose-rate (CONV; 0.07-0.1 Gy⋅s-1) modalities, we showed that FLASH did not cause radiation-induced deficits in learning and memory in mice. Moreover, 6 months after exposure, CONV caused permanent alterations in neurocognitive end points, whereas FLASH did not induce behaviors characteristic of anxiety and depression and did not impair extinction memory. Mechanistic investigations showed that increasing the oxygen tension in the brain through carbogen breathing reversed the neuroprotective effects of FLASH, while radiochemical studies confirmed that FLASH produced lower levels of the toxic reactive oxygen species hydrogen peroxide. In addition, FLASH did not induce neuroinflammation, a process described as oxidative stress-dependent, and was also associated with a marked preservation of neuronal morphology and dendritic spine density. The remarkable normal tissue sparing afforded by FLASH may someday provide heretofore unrealized opportunities for dose escalation to the tumor bed, capabilities that promise to hasten the translation of this groundbreaking irradiation modality into clinical practice.

Enhanced activity and overexpression of Pannexin 1 (Panx1) channels contribute to neuronal pathologies such as epilepsy and Alzheimer's disease (AD). The Panx1 channel ablation alters the hippocampus's glutamatergic neurotransmission, synaptic plasticity, and memory flexibility. Nevertheless, Panx1-knockout (Panx1-KO) mice still retain the ability to learn, suggesting that compensatory mechanisms stabilize their neuronal activity. Here, we show that the absence of Panx1 in the adult brain promotes a series of structural and functional modifications in the Panx1-KO hippocampal synapses, preserving spontaneous activity. Compared to the wild-type (WT) condition, the adult hippocampal neurons of Panx1-KO mice exhibit enhanced excitability, a more complex dendritic branching, enhanced spine maturation, and an increased proportion of multiple synaptic contacts. These modifications seem to rely on the actin–cytoskeleton dynamics as an increase in the actin polymerization and an imbalance between the Rac1 and the RhoA GTPase activities were observed in Panx1-KO brain tissues. Our findings highlight a novel interaction between Panx1 channels, actin, and Rho GTPases, which appear to be relevant for synapse stability. [ABSTRACT FROM AUTHOR]

Creating high-quality polygonal meshes which represent the membrane surface of neurons for both visualization and numerical simulation purposes is an important yet nontrivial task, due to their irregular and complicated structures. In this paper, we develop a novel approach of constructing a watertight 3D mesh from the abstract point-and-diameter representation of the given neuronal morphology. The membrane shape of the neuron is reconstructed by progressively deforming an initial sphere with the guidance of the neuronal skeleton, which can be regarded as a digital sculpting process. To efficiently deform the surface, a local mapping is adopted to simulate the animation skinning. As a result, only the vertices within the region of influence (ROI) of the current skeletal position need to be updated. The ROI is determined based on the finite-support convolution kernel, which is convolved along the line skeleton of the neuron to generate a potential field that further smooths the overall surface at both unidirectional and bifurcating regions. Meanwhile, the mesh quality during the entire evolution is always guaranteed by a set of quasi-uniform rules, which split excessively long edges, collapse undersized ones, and adjust vertices within the tangent plane to produce regular triangles. Additionally, the local vertices density on the result mesh is decided by the radius and curvature of neurites to achieve adaptiveness. [ABSTRACT FROM AUTHOR]

Neuronal plasticity is enhanced in an enriched environment (EE) with more sensory and social interaction. In an animal model of endogenous depression, we have previously shown that EE has positive effects on spatial memory and hippocampus synaptic plasticity. However, nothing is known about how EE influences dendritic remodelling in hippocampal neurons affected by endogenous depression. In depressed rats, the impact of EE on hippocampus neuronal morphology was examined. Neonatal clomipramine exposure from postnatal days (PND) 8-21 days induced endogenous depression. The depressed-like rats were exposed to an enriched environment for two weeks in adulthood. Brains were then collected, stained with a modified Golgi-cox technique and, the hippocampal CA1 dendritic arborisation was evaluated using the Neurolucida software. Depression resulted in the atrophy of CA1 hippocampal neurons. The number of branching points and the overall number of dendritic intersections were reduced in depressed rats,. Exposure to an enriched environment significantly increased dendritic branching and the total number of dendritic intersections in hippocampal CA1 pyramidal neurons. The hippocampal pyramidal neuronal morphology of depressed rats improved after exposure to environmental enrichment. Neuronal plasticity and the development of novel therapeutic strategy will be improved by a greater understanding of how the environment affects neuronal morphology in depressed states. [ABSTRACT FROM AUTHOR]

The lipid phosphatase PTEN (phosphatase and tensin homologue on chromosome 10) is a key tumour suppressor gene and an important regulator of neuronal signalling. PTEN mutations have been identified in patients with autism spectrum disorders, characterized by macrocephaly, impaired social interactions and communication, repetitive behaviour, intellectual disability, and epilepsy. PTEN enzymatic activity is regulated by a cluster of phosphorylation sites at the C-terminus of the protein. Here, we focused on the role of PTEN T366 phosphorylation and generated a knock-in mouse line in which Pten T366 was substituted with alanine (PtenT366A/T366A). We identify that phosphorylation of PTEN at T366 controls neuron size and connectivity of brain circuits involved in sensory processing. We show in behavioural tests that PtenT366/T366A mice exhibit cognitive deficits and selective sensory impairments, with significant differences in male individuals. We identify restricted cellular overgrowth of cortical neurons in PtenT366A/T366A brains, linked to increases in both dendritic arborization and soma size. In a combinatorial approach of anterograde and retrograde monosynaptic tracing using rabies virus, we characterize differences in connectivity to the primary somatosensory cortex of PtenT366A/T366A brains, with imbalances in long-range cortico-cortical input to neurons. We conclude that phosphorylation of PTEN at T366 controls neuron size and connectivity of brain circuits involved in sensory processing and propose that PTEN T366 signalling may account for a subset of autism-related functions of PTEN. [ABSTRACT FROM AUTHOR]

Background: The classical motor symptoms of Parkinson's disease (PD) are tightly linked to the gradual loss of dopamine within the striatum. Concomitantly, medium spiny neurons (MSNs) also experience morphological changes, such as reduced dendritic complexity and spine density, which may be potentially associated with motor dysfunction as well. Thus, MSNs may serve as the emerging targets for PD therapy besides the midbrain dopaminergic neurons. Results: To comprehensively examine pathological alterations of MSNs longitudinally, we established a THCre/Pitx3fl/fl (Pitx3cKO) mouse model that developed canonical PD features, including a significant loss of SNc DAergic neurons and motor deficits. During aging, the targeted neurotransmitter, MSNs morphology and DNA methylation profile were significantly altered upon Pitx3 deficiency. Specifically, dopamine, GABA and glutamate decreased in the model at the early stage. While nuclear, soma and dendritic atrophy, as well as nuclear invaginations increased in the aged MSNs of Pitx3cko mice. Furthermore, more nuclear DNA damages were characterized in MSNs during aging, and Pitx3 deficiency aggravated this phenomenon, together with alterations of DNA methylation profiling associated with lipoprotein and nucleus pathway at the late stage. Conclusion: The early perturbations of the neurotransmitters within MSNs may potentially contribute to the alterations of metabolism, morphology and epigenetics within the striatum at the late stage, which may provide new perspectives on the diagnosis and pathogenesis of PD. [ABSTRACT FROM AUTHOR]

• NMDA receptor antagonist MK801 reduces PyrN spine density and stability. • MK801 does not alter dendritic growth and branching. • Axons that synapse onto PyrN spines exhibit reduced arbor growth and branching in response to MK801 treatment. NMDA-type glutamate receptors play a critical role in activity-dependent neurite growth. We employed cell type-specific genetic labeling in zebrafish to examine the effects of NMDA receptor antagonism on the morphological development of tectal pyramidal neurons (PyrNs). Our data demonstrate that the NMDA receptor antagonist MK801 reduces PyrN spine density and stability without significantly altering dendritic growth and branching. However, the axons that synapse onto PyrN dendritic spines do exhibit reduced arbor growth and branching in response to MK801 treatment. Axons that synapse with PyrNs, but not on spines, are unaffected by MK801 treatment. These findings may reflect different roles for NMDARs during the development of spiny and aspiny dendrites. [ABSTRACT FROM AUTHOR]

Most functions of the nervous system depend on neuronal and glial morphology. Continuous advances in microscopic imaging and tracing software have provided an increasingly abundant availability of 3D reconstructions of arborizing dendrites, axons, and processes, allowing their detailed study. However, efficient, large-scale methods to rank neural morphologies by similarity to an archetype are still lacking. Using the NeuroMorpho.Org database, we present a similarity search software enabling fast morphological comparison of hundreds of thousands of neural reconstructions from any species, brain regions, cell types, and preparation protocols. We compared the performance of different morphological measurements: 1) summary morphometrics calculated by L -Measure, 2) persistence vectors, a vectorized descriptor of branching structure, 3) the combination of the two. In all cases, we also investigated the impact of applying dimensionality reduction using principal component analysis (PCA). We assessed qualitative performance by gauging the ability to rank neurons in order of visual similarity. Moreover, we quantified information content by examining explained variance and benchmarked the ability to identify occasional duplicate reconstructions of the same specimen. We also compared two different methods for selecting the number of principal components using this benchmark. The results indicate that combining summary morphometrics and persistence vectors with applied PCA using maximum likelihood based automatic dimensionality selection provides an information rich characterization that enables efficient and precise comparison of neural morphology. We have deployed the similarity search as open-source online software both through a user-friendly graphical interface and as an API for programmatic access. [Display omitted] • Implemented similarity search enables fast comparison of neuronal reconstructions. • Our method combines summary morphometrics with descriptors of branching structure. • We evaluated visual similarity, information content, and duplicate detection ability. • Similarity search provided both as GUI and API for NeuroMorpho.Org database. • Code and application are released open source for further community development. [ABSTRACT FROM AUTHOR]

Background: Dendritic spines are tiny protrusions found along the dendrites of neurons, and their number is a measure of the density of synaptic connections. Altered density and morphology is observed in several pathologies, and spine formation as well as morphological changes correlate with learning and memory. The detection of spines in microscopy images and the analysis of their morphology is therefore a prerequisite for many studies. We have developed a new open-source, freely available, plugin for ImageJ/FIJI, called Spot Spine, that allows detection and morphological measurements of spines in three dimensional images., Method: Local maxima are detected in spine heads, and the intensity distribution around the local maximum is computed to perform the segmentation of each spine head. Spine necks are then traced from the spine head to the dendrite. Several parameters can be set to optimize detection and segmentation, and manual correction gives further control over the result of the process., Results: The plugin allows the analysis of images of dendrites obtained with various labeling and imaging methods. Quantitative measurements are retrieved including spine head volume and surface, and neck length., Conclusion: The plugin and instructions for use are available at https://imagej.net/plugins/spot-spine., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Gilles JF et al.)

Abstract The digital reconstruction of a neuron is the most direct and effective way to investigate its morphology. Many automatic neuron tracing methods have been proposed, but without manual check it is difficult to know whether a reconstruction or which substructure in a reconstruction is accurate. For a neuron’s reconstructions generated by multiple automatic tracing methods with different principles or models, their common substructures are highly reliable and named individual motifs. In this work, we propose a Vaa3D-based method called Lamotif to explore individual motifs in automatic reconstructions of a neuron. Lamotif utilizes the local alignment algorithm in BlastNeuron to extract local alignment pairs between a specified objective reconstruction and multiple reference reconstructions, and combines these pairs to generate individual motifs on the objective reconstruction. The proposed Lamotif is evaluated on reconstructions of 163 multiple species neurons, which are generated by four state-of-the-art tracing methods. Experimental results show that individual motifs are almost on corresponding gold standard reconstructions and have much higher precision rate than objective reconstructions themselves. Furthermore, an objective reconstruction is mostly quite accurate if its individual motifs have high recall rate. Individual motifs contain common geometry substructures in multiple reconstructions, and can be used to select some accurate substructures from a reconstruction or some accurate reconstructions from automatic reconstruction dataset of different neurons.

In mammals, the zona incerta is thought to be involved in a number of behaviors: visceral activity, arousal, attention, and posture and locomotion. These diverse and complex features suggested that the zona incerta functions as a global or integrative node. Nevertheless, despite multiple investigations into its anatomy, physiology, and behavior in a variety of mammals, no specific character identifies the zona incerta besides its appearance in fiber‐stained material and its relationship to surrounding structures. One such structure is the thalamic reticular nucleus whose caudal pole often contains some intermingled cells of the zona incerta. In crocodilians, the entopeduncular nucleus (ep) abuts the caudal pole of the thalamic reticular nucleus and displays different immunohistochemical properties and soma size when compared with neurons in the thalamic reticular nucleus itself. To determine if neurons in the ep differed from those in the thalamic reticular nucleus in Alligator mississippiensis, the ep was investigated using Golgi methodology. The morphology and soma size of neurons in the ep differed from those in the thalamic reticular nucleus and indicated that these two areas are indeed separate neuronal aggregates. Based on these data and the known relationships of the zona incerta to surrounding structures in mammals, the ep of crocodilians is suggested to be the counterpart of the zona incerta of mammals. [ABSTRACT FROM AUTHOR]

Neuronal morphology influences synaptic connectivity and neuronal signal processing. However, it remains unclear how neuronal shape affects steady-state distributions of organelles like mitochondria. In this work, we investigated the link between mitochondrial transport and dendrite branching patterns by combining mathematical modeling with in vivo measurements of dendrite architecture, mitochondrial motility, and mitochondrial localization patterns in Drosophila HS (horizontal system) neurons. In our model, different forms of morphological and transport scaling rules—which set the relative thicknesses of parent and daughter branches at each junction in the dendritic arbor and link mitochondrial motility to branch thickness—predict dramatically different global mitochondrial localization patterns. We show that HS dendrites obey the specific subset of scaling rules that, in our model, lead to realistic mitochondrial distributions. Moreover, we demonstrate that neuronal activity does not affect mitochondrial transport or localization, indicating that steady-state mitochondrial distributions are hard-wired by the architecture of the neuron. [Display omitted] • A model for transport in dendrites predicts global mitochondrial localization patterns • In this model, specific scaling rules recapitulate realistic mitochondrial distributions • Drosophila HS dendrites obey these specific scaling rules Donovan et al. combine mathematical modeling with in vivo measurements of mitochondrial transport and neuronal architecture in Drosophila HS (horizontal system) neurons to demonstrate that global, steady-state mitochondrial localization patterns depend on specific dendrite branch scaling rules. [ABSTRACT FROM AUTHOR]

Abstract Since manual tracing is time consuming and the performance of automatic tracing is unstable, it is still a challenging task to generate accurate neuron reconstruction efficiently and effectively. One strategy is generating a reconstruction automatically and then amending its inaccurate parts manually. Aiming at finding inaccurate substructures efficiently, we propose a pipeline to retrieve similar substructures on one or more neuron reconstructions, which are very similar to a marked problematic substructure. The pipeline consists of four steps: getting a marked substructure, constructing a query substructure, generating candidate substructures and retrieving most similar substructures. The retrieval procedure was tested on 163 gold standard reconstructions provided by the BigNeuron project and a reconstruction of a mouse’s large neuron. Experimental results showed that the implementation of the proposed methods is very efficient and all retrieved substructures are very similar to the marked one in numbers of nodes and branches, and degree of curvature.

Dynamics of excitable cells and networks depend on the membrane time constant, set by membrane resistance and capacitance. Whereas pharmacological and genetic manipulations of ionic conductances of excitable membranes are routine in electrophysiology, experimental control over capacitance remains a challenge. Here, we present capacitance clamp, an approach that allows electrophysiologists to mimic a modified capacitance in biological neurons via an unconventional application of the dynamic clamp technique. We first demonstrate the feasibility to quantitatively modulate capacitance in a mathematical neuron model and then confirm the functionality of capacitance clamp in in vitro experiments in granule cells of rodent dentate gyrus with up to threefold virtual capacitance changes. Clamping of capacitance thus constitutes a novel technique to probe and decipher mechanisms of neuronal signaling in ways that were so far inaccessible to experimental electrophysiology.

The ventral pallidum (VP) integrates reward signals to regulate cognitive, emotional, and motor processes associated with motivational salience. Previous studies have revealed that the VP projects axons to many cortical and subcortical structures. However, descriptions of the neuronal morphologies and projection patterns of the VP neurons at the single neuron level are lacking, thus hindering the understanding of the wiring diagram of the VP. In this study, we used recently developed progress in robust sparse labeling and fluorescence micro-optical sectioning tomography imaging system (fMOST) to label mediodorsal thalamus-projecting neurons in the VP and obtain high-resolution whole-brain imaging data. Based on these data, we reconstructed VP neurons and classified them into three types according to their fiber projection patterns. We systematically compared the axonal density in various downstream centers and analyzed the soma distribution and dendritic morphologies of the various subtypes at the single neuron level. Our study thus provides a detailed characterization of the morphological features of VP neurons, laying a foundation for exploring the neural circuit organization underlying the important behavioral functions of VP. [ABSTRACT FROM AUTHOR]

The thalamic reticular nucleus (TRN) is a critical structure influencing information transfer to the forebrain. In crocodilians, the TRN shares many features with its mammalian counterpart. One area that has not been explored is how individual neurons in the crocodilian TRN compare with those found in mammals. In mammals, TRN neurons are aligned parallel to the external border of the dorsal thalamus, have their dendrites oriented perpendicular to the fibers in the internal capsule, have fine, filamentous dendritic appendages, are either bipolar or multipolar, and are commonly considered to be a homogeneous morphological population of cells. To investigate the cellular morphology of the TRN complex, a Golgi analysis was undertaken in Alligator mississippiensis. This study examined features that have been used in mammals. In Alligator, the four TRN divisions are the dorsal peduncular nucleus, the perireticular nucleus, the interstitial nucleus, and the neurons in the medial forebrain bundle associated with the interstitial nucleus. In crocodilians, the dorsal peduncular nucleus is homologous to the TRN of mammals. From the 1787 drawn neuron profiles in the traditional three planes of section, the following were concluded. First, neurons in each part of the TRN complex in Alligator were similar in morphology. Second, each part of the TRN complex of Alligator contained a heterogenous population of cells. These variations between the cellular morphology of the dorsal peduncular nucleus of crocodilians and the TRN of mammals are speculated to partly result from differences in forebrain organization. [ABSTRACT FROM AUTHOR]

Abstract Background Neurons are the basic structural unit of the brain, and their morphology is a key determinant of their classification. The morphology of a neuronal circuit is a fundamental component in neuron modeling. Recently, single-neuron morphologies of the whole brain have been used in many studies. The correctness and completeness of semimanually traced neuronal morphology are credible. However, there are some inaccuracies in semimanual tracing results. The distance between consecutive nodes marked by humans is very long, spanning multiple voxels. On the other hand, the nodes are marked around the centerline of the neuronal fiber, not on the centerline. Although these inaccuracies do not seriously affect the projection patterns that these studies focus on, they reduce the accuracy of the traced neuronal skeletons. These small inaccuracies will introduce deviations into subsequent studies that are based on neuronal morphology files. Results We propose a neuronal digital skeleton optimization method to evaluate and make fine adjustments to a digital skeleton after neuron tracing. Provided that the neuronal fiber shape is smooth and continuous, we describe its physical properties according to two shape restrictions. One restriction is designed based on the grayscale image, and the other is designed based on geometry. These two restrictions are designed to finely adjust the digital skeleton points to the neuronal fiber centerline. With this method, we design the three-dimensional shape restriction workflow of neuronal skeleton adjustment computation. The performance of the proposed method has been quantitatively evaluated using synthetic and real neuronal image data. The results show that our method can reduce the difference between the traced neuronal skeleton and the centerline of the neuronal fiber. Furthermore, morphology metrics such as the neuronal fiber length and radius become more precise. Conclusions This method can improve the accuracy of a neuronal digital skeleton based on traced results. The greater the accuracy of the digital skeletons that are acquired, the more precise the neuronal morphologies that are analyzed will be.

Abstract Research advancements in neuroscience entail the production of a substantial amount of data requiring interpretation, analysis, and integration. The complexity and diversity of neuroscience data necessitate the development of specialized databases and associated standards and protocols. NeuroMorpho.Org is an online repository of over one hundred thousand digitally reconstructed neurons and glia shared by hundreds of laboratories worldwide. Every entry of this public resource is associated with essential metadata describing animal species, anatomical region, cell type, experimental condition, and additional information relevant to contextualize the morphological content. Until recently, the lack of a user-friendly, structured metadata annotation system relying on standardized terminologies constituted a major hindrance in this effort, limiting the data release pace. Over the past 2 years, we have transitioned the original spreadsheet-based metadata annotation system of NeuroMorpho.Org to a custom-developed, robust, web-based framework for extracting, structuring, and managing neuroscience information. Here we release the metadata portal publicly and explain its functionality to enable usage by data contributors. This framework facilitates metadata annotation, improves terminology management, and accelerates data sharing. Moreover, its open-source development provides the opportunity of adapting and extending the code base to other related research projects with similar requirements. This metadata portal is a beneficial web companion to NeuroMorpho.Org which saves time, reduces errors, and aims to minimize the barrier for direct knowledge sharing by domain experts. The underlying framework can be progressively augmented with the integration of increasingly autonomous machine intelligence components.

The ventral pallidum (VP) integrates reward signals to regulate cognitive, emotional, and motor processes associated with motivational salience. Previous studies have revealed that the VP projects axons to many cortical and subcortical structures. However, descriptions of the neuronal morphologies and projection patterns of the VP neurons at the single neuron level are lacking, thus hindering the understanding of the wiring diagram of the VP. In this study, we used recently developed progress in robust sparse labeling and fluorescence micro-optical sectioning tomography imaging system (fMOST) to label mediodorsal thalamus-projecting neurons in the VP and obtain high-resolution whole-brain imaging data. Based on these data, we reconstructed VP neurons and classified them into three types according to their fiber projection patterns. We systematically compared the axonal density in various downstream centers and analyzed the soma distribution and dendritic morphologies of the various subtypes at the single neuron level. Our study thus provides a detailed characterization of the morphological features of VP neurons, laying a foundation for exploring the neural circuit organization underlying the important behavioral functions of VP.

Enhanced activity and overexpression of Pannexin 1 (Panx1) channels contribute to neuronal pathologies such as epilepsy and Alzheimer’s disease (AD). The Panx1 channel ablation alters the hippocampus’s glutamatergic neurotransmission, synaptic plasticity, and memory flexibility. Nevertheless, Panx1-knockout (Panx1-KO) mice still retain the ability to learn, suggesting that compensatory mechanisms stabilize their neuronal activity. Here, we show that the absence of Panx1 in the adult brain promotes a series of structural and functional modifications in the Panx1-KO hippocampal synapses, preserving spontaneous activity. Compared to the wild-type (WT) condition, the adult hippocampal neurons of Panx1-KO mice exhibit enhanced excitability, a more complex dendritic branching, enhanced spine maturation, and an increased proportion of multiple synaptic contacts. These modifications seem to rely on the actin–cytoskeleton dynamics as an increase in the actin polymerization and an imbalance between the Rac1 and the RhoA GTPase activities were observed in Panx1-KO brain tissues. Our findings highlight a novel interaction between Panx1 channels, actin, and Rho GTPases, which appear to be relevant for synapse stability.

Adult neural plasticity is an important and intriguing phenomenon in the brain, and adult hippocampal neurogenesis is directly involved in modulating neural plasticity by mechanisms that are only partially understood. We have performed gainof-function and loss-of-function experiments to study Smad2, a transcription factor selected from genes that are demethylated after exercise through the analysis of an array of physical activity-induced factors, and their corresponding gene expression, and an efficient inducer of plasticity. In these studies, changes in cell number and morphology were analyzed in the hippocampal dentate gyrus (cell proliferation and survival, including regional distribution, and structural maturation/differentiation, including arborization, dendritic spines, and neurotransmitter-specific vesicles) of sedentary male mice, after evaluation in a battery of behavioral tests. As a result, we reveal a role for Smad2 in the balance of proliferation versus maturation of differentiating immature cells (Smad2 silencing increases both the proliferation and survival of cycling cells in the dentate granule cell layer), and in the plasticity of both newborn and mature neurons in mice (by decreasing dendritic arborization and dendritic spine number). Moreover, Smad2 silencing specifically compromises spatial learning in mice (through impairments of spatial tasks acquisition both in long-term learning and working memory). These data suggest that Smad2 participates in adult neural plasticity by influencing the proliferation and maturation of dentate gyrus neurons. [ABSTRACT FROM AUTHOR]