The Effects of Acute GABA Treatment on the Functional Connectivity and Network Topology of Cortical Cultures.

γ-Aminobutyric acid (GABA) is an inhibitory transmitter, acting on receptor channels to reduce neuronal excitability in matured neural systems. However, electrophysiological responses of whole neuronal ensembles to the exposure to GABA are still unclear. We used micro-electrode arrays (MEAs) to study the effects of the increasing amount of GABA on functional network of cortical neural cultures. Then the recorded data were analyzed by the cross-covariance analysis and graph theory. Results showed that after the GABA treatment, the activity parameters of firing rate, bursting rate, bursting duration and network burst frequency in neural cultures decreased as expected. In addition, the functional connectivity also decreased in similarity, network density, and the size of the largest component. However, small-worldness was not found to be influenced by the acute GABA treatment. Our results support the position that using graph theory to evaluate the functional connectivity of neural cultures may enhance understanding of the pharmacological impact of neurotransmitters on neuronal networks.

Loss of SIL1 Affects Actin Dynamics and Leads to Abnormal Neural Migration.

SIL1 is a nucleotide exchange factor for the molecular chaperone protein Bip in the endoplasmic reticulum that plays a crucial role in protein folding. The Sil1 gene is currently the only known causative gene of Marinesco-Sjögren syndrome (MSS). Intellectual developmental disability is the main symptom of MSS, and its mechanism has not been fully elucidated. Studies have shown that mutations in the Sil1 gene can delay neuronal migration during cortical development, but the underlying molecular mechanisms remain unclear. To further identify potential molecules involved in the regulation of central nervous system development by SIL1, we established a cortical neuron model with SIL1 protein deficiency and used proteomic analysis to screen for differentially expressed proteins after Sil1 silencing, followed by GO functional enrichment and protein‒protein interaction (PPI) network analysis. We identified 68 upregulated and 137 downregulated proteins in total, and among them, 10 upregulated and 3 downregulated proteins were mainly related to actin cytoskeleton dynamics. We further validated the differential changes in actin-related molecules using qRT‒PCR and Western blotting of a Sil1 gene knockout (Sil1-/-) mouse model. The results showed that the protein levels of ACTN1 and VIM decreased, while their mRNA levels increased as a compensatory response to protein deficiency. The mRNA and protein levels of IQGAP1 both showed a secondary increase. In conclusion, we identified ACTN1 and VIM as the key molecules regulated by SIL1 that are involved in neuronal migration during cortical development.

Shape-changing electrode array for minimally invasive large-scale intracranial brain activity mapping.

Large-scale brain activity mapping is important for understanding the neural basis of behaviour. Electrocorticograms (ECoGs) have high spatiotemporal resolution, bandwidth, and signal quality. However, the invasiveness and surgical risks of electrode array implantation limit its application scope. We developed an ultrathin, flexible shape-changing electrode array (SCEA) for large-scale ECoG mapping with minimal invasiveness. SCEAs were inserted into cortical surfaces in compressed states through small openings in the skull or dura and fully expanded to cover large cortical areas. MRI and histological studies on rats proved the minimal invasiveness of the implantation process and the high chronic biocompatibility of the SCEAs. High-quality micro-ECoG activities mapped with SCEAs from male rodent brains during seizures and canine brains during the emergence period revealed the spatiotemporal organization of different brain states with resolution and bandwidth that cannot be achieved using existing noninvasive techniques. The biocompatibility and ability to map large-scale physiological and pathological cortical activities with high spatiotemporal resolution, bandwidth, and signal quality in a minimally invasive manner offer SCEAs as a superior tool for applications ranging from fundamental brain research to brain-machine interfaces.

A high-density 1,024-channel probe for brain-wide recordings in non-human primates.

Large-scale neural population recordings with single-cell resolution across the primate brain remain challenging. Here we introduce the Neuroscroll probe that isolates single neuronal activities simultaneously from 1,024 densely spaced channels that are flexibly distributed across the shank of the probe. The Neuroscroll probe length is easily tunable for individual probes from 10 mm to 90 mm, covering the brain size of non-human primates and humans, and the probes remain intact and functional after repeated bending deformations. The Neuroscroll probes provided reliable recordings from large neural populations with high chronic stability up to 105 weeks in rats. Recording with each Neuroscroll probe yielded hundreds of well-isolated single units simultaneously from multiple brain regions distributed across the entire depth of the rhesus macaque brain. With the thousand simultaneously recorded channels, unprecedented probe length, excellent mechanical stability and flexible recording site distribution, the Neuroscroll probes enable a wide range of new experimental paradigms in system neuroscience studies with great versatility.

Overexpression of soybean GmDHN9 gene enhances drought resistance of transgenic Arabidopsis.

Soybean is one of the important oil crops and a major source of protein and lipids. Drought can cause severe soybean yields. Dehydrin protein (DHN) is a subfamily of LEA proteins that play an important role in plant responses to abiotic stresses. In this study, the soybean GmDHN9 gene was cloned and induced under a variety of abiotic stresses. Results showed that the GmDHN9 gene response was more pronounced under drought induction. Subcellular localization results indicated that the protein was localized in the cytoplasm. The role of transgenic Arabidopsis plants in drought stress response was further studied. Under drought stress, the germination rate, root length, chlorophyll, proline, relative water content, and antioxidant enzyme content of transgenic Arabidopsis thaliana transgenic genes were higher than those of wild-type plants, and transgenic plants contained less O2-, H2O2 and MDA contents. In short, the GmDHN9 gene can regulate the homeostasis of ROS and enhance the drought resistance of plants.

Macaque Brainnetome Atlas: A multifaceted brain map with parcellation, connection, and histology.

The rhesus macaque (Macaca mulatta) is a crucial experimental animal that shares many genetic, brain organizational, and behavioral characteristics with humans. A macaque brain atlas is fundamental to biomedical and evolutionary research. However, even though connectivity is vital for understanding brain functions, a connectivity-based whole-brain atlas of the macaque has not previously been made. In this study, we created a new whole-brain map, the Macaque Brainnetome Atlas (MacBNA), based on the anatomical connectivity profiles provided by high angular and spatial resolution ex vivo diffusion MRI data. The new atlas consists of 248 cortical and 56 subcortical regions as well as their structural and functional connections. The parcellation and the diffusion-based tractography were evaluated with invasive neuronal-tracing and Nissl-stained images. As a demonstrative application, the structural connectivity divergence between macaque and human brains was mapped using the Brainnetome atlases of those two species to uncover the genetic underpinnings of the evolutionary changes in brain structure. The resulting resource includes: (1) the thoroughly delineated Macaque Brainnetome Atlas (MacBNA), (2) regional connectivity profiles, (3) the postmortem high-resolution macaque diffusion and T2-weighted MRI dataset (Brainnetome-8), and (4) multi-contrast MRI, neuronal-tracing, and histological images collected from a single macaque. MacBNA can serve as a common reference frame for mapping multifaceted features across modalities and spatial scales and for integrative investigation and characterization of brain organization and function. Therefore, it will enrich the collaborative resource platform for nonhuman primates and facilitate translational and comparative neuroscience research.

High-Performance Artificial Ligament Made from Helical Polyester Fibers Wrapped with Aligned Carbon Nanotube Sheets.

Repairing high-load connective tissues, such as ligaments, by surgically implanting artificial grafts after injury is challenging because they lack biointegration with host bones for stable interfaces. Herein, a high-performance helical composite fiber (HCF) ligament by wrapping aligned carbon nanotube (CNT) sheets around polyester fibers is proposed. Anterior cruciate ligament (ACL) reconstruction surgery shows that HCF grafts could induce effective bone regeneration, thus allowing the narrowing of bone tunnel defects. Such repair of the bone tunnel is in strong contrast to the tunnel enlargement of more than 50% for commercial artificial ligaments made from bare polyester fibers. Rats reconstructed with this HCF ligament show normal jumping, walking, and running without limping. This work allows bone regeneration in vivo through a one-step surgery without seeding cells or transforming growth factors, thereby opening an avenue for high-performance artificial tissues.

The regulatory role of endoplasmic reticulum chaperone proteins in neurodevelopment.

The endoplasmic reticulum (ER) is the largest tubular reticular organelle spanning the cell. As the main site of protein synthesis, Ca2+ homeostasis maintenance and lipid metabolism, the ER plays a variety of essential roles in eukaryotic cells, with ER molecular chaperones participate in all these processes. In recent years, it has been reported that the abnormal expression of ER chaperones often leads to a variety of neurodevelopmental disorders (NDDs), including abnormal neuronal migration, neuronal morphogenesis, and synaptic function. Neuronal development is a complex and precisely regulated process. Currently, the mechanism by which neural development is regulated at the ER level remains under investigation. Therefore, in this work, we reviewed the recent advances in the roles of ER chaperones in neural development and developmental disorders caused by the deficiency of these molecular chaperones.

A Computational Model to Investigate GABA-Activated Astrocyte Modulation of Neuronal Excitation.

Gamma-aminobutyric acid (GABA) is critical for proper neural network function and can activate astrocytes to induce neuronal excitability; however, the mechanism by which astrocytes transform inhibitory signaling to excitatory enhancement remains unclear. Computational modeling can be a powerful tool to provide further understanding of how GABA-activated astrocytes modulate neuronal excitation. In the present study, we implemented a biophysical neuronal network model to investigate the effects of astrocytes on excitatory pre- and postsynaptic terminals following exposure to increasing concentrations of external GABA. The model completely describes the effects of GABA on astrocytes and excitatory presynaptic terminals within the framework of glutamatergic gliotransmission according to neurophysiological findings. Utilizing this model, our results show that astrocytes can rapidly respond to incoming GABA by inducing Ca2+ oscillations and subsequent gliotransmitter glutamate release. Elevation in GABA concentrations not only naturally decreases neuronal spikes but also enhances astrocytic glutamate release, which leads to an increase in astrocyte-mediated presynaptic release and postsynaptic slow inward currents. Neuronal excitation induced by GABA-activated astrocytes partly counteracts the inhibitory effect of GABA. Overall, the model helps to increase knowledge regarding the involvement of astrocytes in neuronal regulation using simulated bath perfusion of GABA, which may be useful for exploring the effects of GABA-type antiepileptic drugs.

First molecular evidence of Anaplasma spp. co-infection in stray dogs from Anhui, China.

Anaplasma species are obligate intracellular pathogens that threaten the health of both humans and animals worldwide. This study aimed to determine the prevalence of Anaplasma species in stray dogs in Anhui Province, China. Blood samples from 201 apparently healthy stray dogs were collected from August 2017 to January 2018, and Anaplasma spp. infection in these dogs was evaluated by nested PCR and phylogenetic analysis. The overall infection rate of Anaplasma spp. in stray dogs was 38.3% (77/201). The prevalences of single infection of A. platys, A. phagocytophilum and A. ovis were 15.4%, 11.9%, and 8.5%, respectively. Co-infection rate of A. platys and A. phagocytophilum was 1.5% and that of A. platys and A. ovis was 0.5%. Co-infection by these three pathogens was found in one sample (0.5%). This is the first report of Anaplasma spp. infections in stray dogs from Anhui, China.

The effect of acute glutamate treatment on the functional connectivity and network topology of cortical cultures.

Microelectrode arrays (MEAs) allow the investigation of the pharmacological and toxicological effects of chemicals on cultured neuronal networks. Understanding the functional connections between neurons and the resulting neuronal networks is important for evaluating drugs that affect synaptic transmission. Therefore, we acutely treated a mature cultured neuronal network on MEAs with accumulating amounts of glutamate and recorded their altered electrophysiology. Subsequently, a cross-covariance analysis was applied to process the spiking activity in the network and to evaluate the connections between neurons. Finally, graph theory was used to assess the functional network properties under acute glutamate treatment. Our data demonstrated that glutamate increased the similarity, connectivity weight, density, and largest-component size of the functional network. In addition, the small-world network topology was altered after glutamate treatment. Our results indicate that the graph theory can advance our understanding of the pharmacological significance of neurotransmitters on neuronal networks.

Reliability of motor and sensory neural decoding by threshold crossings for intracortical brain-machine interface.

OBJECTIVE: For intracortical neurophysiological studies, spike sorting is an important procedure to isolate single units for analyzing specific functions. However, whether spike sorting is necessary or not for neural decoding applications is controversial. Several studies showed that using threshold crossings (TC) instead of spike sorting could also achieve a similar satisfactory performance. However, such studies were limited in similar behavioral tasks, and the neural signal source mainly focused on the motor-related cortical regions. It is not certain if this conclusion is applicable to other situations. Therefore, we compared the performance of TC and spike sorting in neural decoding with more comprehensive paradigms and parameters. APPROACH: Two rhesus macaques implanted with Utah or floating microelectrode arrays (FMAs) in motor or sensory-related cortical regions were trained to perform a motor or a sensory task. Data from each monkey were preprocessed with three different schemes: TC, automatic sorting (AS), and manual sorting (MS). A support vector machine was used as the decoder, and the decoding accuracy was used for evaluating the performance of three preprocessing methods. Different neural signal sources, different decoders, and related parameters and decoding stability were further tested to systematically compare three preprocessing methods. MAIN RESULTS: TC could achieve a similar (-4.5 RMS threshold) or better (-3.0 RMS threshold) decoding performance compared to the other two sorting methods in the motor or sensory tasks even if the neural signal sources or decoder-related parameters were changed. Moreover, TC was much more stable in neural decoding across sessions and robust to changes of threshold. SIGNIFICANCE: Our results indicated that spike-firing patterns could be stably extracted through TC from multiple cortices in both motor and sensory neural decoding applications. Considering the stability of TC, it might be more suitable for neural decoding compared to sorting methods.

Modulation of Beta Oscillations for Implicit Motor Timing in Primate Sensorimotor Cortex during Movement Preparation.

Motor timing is an important part of sensorimotor control. Previous studies have shown that beta oscillations embody the process of temporal perception in explicit timing tasks. In contrast, studies focusing on beta oscillations in implicit timing tasks are lacking. In this study, we set up an implicit motor timing task and found a modulation pattern of beta oscillations with temporal perception during movement preparation. We trained two macaques in a repetitive visually-guided reach-to-grasp task with different holding intervals. Spikes and local field potentials were recorded from microelectrode arrays in the primary motor cortex, primary somatosensory cortex, and posterior parietal cortex. We analyzed the association between beta oscillations and temporal interval in fixed-duration experiments (500 ms as the Short Group and 1500 ms as the Long Group) and random-duration experiments (500 ms to 1500 ms). The results showed that the peak beta frequencies in both experiments ranged from 15 Hz to 25 Hz. The beta power was higher during the hold period than the movement (reach and grasp) period. Further, in the fixed-duration experiments, the mean power as well as the maximum rate of change of beta power in the first 300 ms were higher in the Short Group than in the Long Group when aligned with the Center Hit event. In contrast, in the random-duration experiments, the corresponding values showed no statistical differences among groups. The peak latency of beta power was shorter in the Short Group than in the Long Group in the fixed-duration experiments, while no consistent modulation pattern was found in the random-duration experiments. These results indicate that beta oscillations can modulate with temporal interval in their power mode. The synchronization period of beta power could reflect the cognitive set maintaining working memory of the temporal structure and attention.

Influence of Heat Treatments on Microstructure and Mechanical Properties of Ti⁻26Nb Alloy Elaborated In Situ by Laser Additive Manufacturing with Ti and Nb Mixed Powder.

In the present work, a Ti⁻26Nb alloy was elaborated in situ by laser additive manufacturing (LAM) with Ti and Nb mixed powders. The alloys were annealed at temperatures ranging from 650 °C to 925 °C, and the effects of the annealing temperature on the microstructure and mechanical properties were investigated. It has been found that the microstructure of the as-deposited alloy obtained in the present conditions is characterized by columnar prior ß grains with a relatively strong <001> fiber texture in the build direction. The as-deposited alloy exhibits extremely high strength, and its ultimate tensile strength and yield strength are about 799 MPa and 768 MPa, respectively. The annealing temperature has significant effects on the microstructure and mechanical properties of the alloys. Annealing treatment can promote the dissolution of unmelted Nb particles and eliminate the micro-segregation of Nb at the elliptical-shaped grain boundaries, while increasing the grain size of the alloy. With an increase in annealing temperature, the strength of the alloy decreases but the ductility increases. The alloy annealed at 850 °C exhibits a balance of strength and ductility.

A Fast, Open EEG Classification Framework Based on Feature Compression and Channel Ranking.

Superior feature extraction, channel selection and classification methods are essential for designing electroencephalography (EEG) classification frameworks. However, the performance of most frameworks is limited by their improper channel selection methods and too specifical design, leading to high computational complexity, non-convergent procedure and narrow expansibility. In this paper, to remedy these drawbacks, we propose a fast, open EEG classification framework centralized by EEG feature compression, low-dimensional representation, and convergent iterative channel ranking. First, to reduce the complexity, we use data clustering to compress the EEG features channel-wise, packing the high-dimensional EEG signal, and endowing them with numerical signatures. Second, to provide easy access to alternative superior methods, we structurally represent each EEG trial in a feature vector with its corresponding numerical signature. Thus, the recorded signals of many trials shrink to a low-dimensional structural matrix compatible with most pattern recognition methods. Third, a series of effective iterative feature selection approaches with theoretical convergence is introduced to rank the EEG channels and remove redundant ones, further accelerating the EEG classification process and ensuring its stability. Finally, a classical linear discriminant analysis (LDA) model is employed to classify a single EEG trial with selected channels. Experimental results on two real world brain-computer interface (BCI) competition datasets demonstrate the promising performance of the proposed framework over state-of-the-art methods.

Improving Generalization Based on l1-Norm Regularization for EEG-Based Motor Imagery Classification.

Multichannel electroencephalography (EEG) is widely used in typical brain-computer interface (BCI) systems. In general, a number of parameters are essential for a EEG classification algorithm due to redundant features involved in EEG signals. However, the generalization of the EEG method is often adversely affected by the model complexity, considerably coherent with its number of undetermined parameters, further leading to heavy overfitting. To decrease the complexity and improve the generalization of EEG method, we present a novel l1-norm-based approach to combine the decision value obtained from each EEG channel directly. By extracting the information from different channels on independent frequency bands (FB) with l1-norm regularization, the method proposed fits the training data with much less parameters compared to common spatial pattern (CSP) methods in order to reduce overfitting. Moreover, an effective and efficient solution to minimize the optimization object is proposed. The experimental results on dataset IVa of BCI competition III and dataset I of BCI competition IV show that, the proposed method contributes to high classification accuracy and increases generalization performance for the classification of MI EEG. As the training set ratio decreases from 80 to 20%, the average classification accuracy on the two datasets changes from 85.86 and 86.13% to 84.81 and 76.59%, respectively. The classification performance and generalization of the proposed method contribute to the practical application of MI based BCI systems.

Injectable OPF/graphene oxide hydrogels provide mechanical support and enhance cell electrical signaling after implantation into myocardial infarct.

After myocardial infarction (MI), the scar tissue contributes to ventricular dysfunction by electrically uncoupling viable cardiomyocytes in the infarct region. Injection of a conductive hydrogel could not only provide mechanical support to the infarcted region, but also synchronize contraction and restore ventricular function by electrically connecting isolated cardiomyocytes to intact tissue. Methods: We created a conductive hydrogel by introducing graphene oxide (GO) nanoparticles into oligo(poly(ethylene glycol) fumarate) (OPF) hydrogels. The hydrogels were characterized by AFM and electrochemistry workstation. A rat model of myocardial infarction was used to investigate the ability of OPF/GO to improve cardiac electrical propagation in the injured heart in vivo. Echocardiography (ECHO) was used to evaluate heart function 4 weeks after MI. Ca2+ imaging was used to visualize beating cardiomyocytes (CMs). Immunofluorescence staining was used to visualize the expression of cardiac-specific markers. Results: OPF/GO hydrogels had semiconductive properties that were lacking in pure OPF. In addition, the incorporation of GO into OPF hydrogels could improve cell attachment in vitro. Injection of OPF/GO 4 weeks after myocardial infarction in rats enhanced the Ca2+ signal conduction of cardiomyocytes in the infarcted region in comparison with PBS or OPF alone. Moreover, the injection of OPF/GO hydrogel into the infarct region enhanced the generation of cytoskeletal structure and intercalated disc assembly. Echocardiography analysis showed improvement in load-dependent ejection fraction/fractional shortening of heart function 4 weeks after injection. Conclusions: We prepared a conductive hydrogel (OPF/GO) that provide mechanical support and biological conduction in vitro and in vivo. We found that injected OPF/GO hydrogels can provide mechanical support and electric connection between healthy myocardium and the cardiomyocytes in the scar via activating the canonical Wnt signal pathway, thus upregulating the generation of Cx43 and gap junction associated proteins. Injection of OPF/GO hydrogel maintained better heart function after myocardial infarction than the injection of a nonconductive polymer.

The complete mitochondrial genome of the greater green snake Cyclophiops major (Reptilia, Serpentes, Colubridae).

The greater green snake Cyclophiops major is a protected and colubrid species. Here, we investigated the complete mitochondrial genome of C. major. The genome is 17,217bp in size, including 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, 2 control regions, and an origin of light-strand replication. All genes are distributed on the heavy strand, except for ND6 gene and 8 tRNA genes. The AT content of the overall base composition of light strand is 59.83%, showing AT bias. Phylogenetic tree was built based on the genome of C. major and other related snakes to analyze their phylogenic relationship.

Synthetic Engineering of Spider Silk Fiber as Implantable Optical Waveguides for Low-Loss Light Guiding.

A variety of devices used for biomedical engineering have been fabricated using protein polymer because of their excellent properties, such as strength, toughness, biocompatibility, and biodegradability. In this study, we fabricated an optical waveguide using genetically engineered spider silk protein. This method has two significant advantages: (1) recombinant spider silk optical waveguide exhibits excellent optical and biological properties and (2) biosynthesis of spider silk protein can overcome the limitation to the research on spider silk optical waveguide due to the low yield of natural spider silk. In detail, two kinds of protein-based optical waveguides made from recombinant spider silk protein and regenerative silkworm silk protein were successfully prepared. Results suggested that the recombinant spider silk optical waveguide showed a smoother surface and a higher refractive index when compared with regenerative silkworm silk protein. The optical loss of recombinant spider silk optical waveguide was 0.8 ± 0.1 dB/cm in air and 1.9 ± 0.3 dB/cm in mouse muscles, which were significantly lower than those of regenerative silkworm silk optical waveguide. Moreover, recombinant spider silk optical waveguide can meet the demand to guide and efficiently deliver light through biological tissue. In addition, recombinant spider silk optical waveguide showed low toxicity to cells in vitro and low-level inflammatory reaction with surrounding tissue in vivo. Therefore, recombinant spider silk optical waveguide is a promising implantable device to guide and deliver light with low loss.

Efficient GSH delivery using PAMAM-GSH into MPP-induced PC12 cellular model for Parkinson's disease.

Glutathione (GSH) depletion has been an important contributor to the dysfunction of dopamine neurons. Polyamidoamine-GSH (PAMAM-GSH) was synthesized and the delivery effect of GSH into PC12 cells was tested. MTT assessment for cytotoxicity and reactive oxygen species (ROS) as well as nitrite oxide (NO) and intracelluar superoxide dismutase (SOD) detection for antioxidative ability were performed. Furthermore, the antiapoptotic ability was analysed by assessing caspase-3, JNK1/2 and Erk1/2 expression. Our data indicated that PAMAM-GSH is an effective agent to replenish GSH into PC12 cells. PAMAM-GSH developed its antioxidative and protective ability for 1-methyl-4-phenylpyridinium (MPP)-induced PC12 cells by reducing the intracellular levels of ROS and SOD activity as well as decreasing the release of NO. Meanwhile, PAMAM-GSH could inhibit caspase-3 activation and might show its antiapoptotic ability to MPP-induced PC12 cells through JNK2/Erk1/2 pathway. In summary, these studies suggest that PAMAM-GSH conjugate has an intrinsic ability to penetrate PC12 cells and deliver GSH into these cells which may provide a new strategy for clinical applications in the treatment of Parkinson's disease.