Tool In Neuroscience Research Research Articles

Viral Tools for Neural Circuit Tracing.

Neural circuits provide an anatomical basis for functional networks. Therefore, dissecting the structure of neural circuits is essential to understanding how the brain works. Recombinant neurotropic viruses are important tools for neural circuit tracing with many advantages over non-viral tracers: they allow for anterograde, retrograde, and trans-synaptic delivery of tracers in a cell type-specific, circuit-selective manner. In this review, we summarize the recent developments in the viral tools for neural circuit tracing, discuss the key principles of using viral tools in neuroscience research, and highlight innovations for developing and optimizing viral tools for neural circuit tracing across diverse animal species, including nonhuman primates.

An Ultra-Low-Noise, Low Power and Miniaturized Dual-Channel Wireless Neural Recording Microsystem.

As the basic tools for neuroscience research, invasive neural recording devices can obtain high-resolution neuronal activity signals through electrodes connected to the subject’s brain. Existing wireless neural recording devices are large in size or need external large-scale equipment for wireless power supply, which limits their application. Here, we developed an ultra-low-noise, low power and miniaturized dual-channel wireless neural recording microsystem. With the full-differential front-end structure of the dual operational amplifiers (op-amps), the noise level and power consumption are notably reduced. The hierarchical microassembly technology, which integrates wafer-level packaged op-amps and the miniaturized Bluetooth module, dramatically reduces the size of the wireless neural recording microsystem. The microsystem shows a less than 100 nV/ ultra-low noise level, about 10 mW low power consumption, and 9 × 7 × 5 mm small size. The neural recording ability was then demonstrated in saline and a chronic rat model. Because of its miniaturization, it can be applied to freely behaving small animals, such as rats. Its features of ultra-low noise and high bandwidth are conducive to low-amplitude neural signal recording, which may help advance neuroscientific discovery.

(Digital Presentation) Bioelectrochemistry of Neuronal Tau Protein

Tau is a neuronal protein and one of the biomarkers of neurodegenerative diseases, such as Alzheimer’s disease. Tau protein undergoes post-translational modifications, aggregation, and is a viable drug target. In addition, this protein is a vital biomarker in biological fluids towards early detection of neurodegenerative diseases. We reported on using electrochemical impedance spectroscopy and cyclic voltammetry for detection of protein-protein interactions (1), protein-ligand interactions (2), enzymatic catalysis and enzyme inhibition (3) while focusing on tau protein and its biochemistry. For example, tau-tau, tau-ferritin, tau-transferrin protein interactions were monitored, and protein kinase-catalyzed phosphorylation of tau protein was detected. The phosphorylation inhibitors, such as antibodies, were screened for their efficacy. Bioelectrochemical methodologies were used to gain insight into various facets of protein biochemistry, and represent promising tools in neuroscience research. References A) Carlin, N., Martic-Milne, S. (2018). Anti-tau antibodies based electrochemical sensor for detection of tau protein. J. Electrochem. Soc.165: G3018-G3025. B) Jahshan, A., Esteves, J.O.V., Martic-Milne, S. (2016). Evaluation of ferritin and transferrin binding to tau protein. J. Inorg. Biochem. 162: 127-134. C) Esteves, J.O.V., Trzeciakiewicz, H., Loeffler, D.A., Martic, S. (2015). Effects of tau domain-specific antibodies and intravenous immunoglobulin on tau aggregation and aggregate degradation. Biochemistry. 54: 15-18.Trzeciakiewicz, H., Esteves, J.O.V., Carlin, N., Martic, S. (2015). Electrochemistry of heparin binding to tau protein on Au surfaces. Electrochim. Acta. 162: 24-30,Esteves, J.O.V., Martic-Milne, S. (2016). Electrochemical detection of anti-tau antibodies binding to tau protein and inhibition of GSK-3-β-catalyzed phosphorylation. Anal. Biochem. 496: 55-62

Behavioral and slice electrophysiological assessment of DREADD ligand, deschloroclozapine (DCZ) in rats

Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) have become a premier neuroscience research tool for enabling reversible manipulations of cellular activity following experimenter-controlled delivery of a DREADD-specific ligand. However, several DREADD ligands, e.g., clozapine-N-oxide (CNO), have metabolic and off-target effects that may confound experimental findings. New DREADD ligands aim to reduce metabolic and potential off-target effects while maintaining strong efficacy for the designer receptors. Recently a novel DREADD ligand, deschloroclozapine (DCZ), was shown to induce chemogenetic-mediated cellular and behavioral effects in mice and monkeys without detectable side effects. The goal of the present study was to examine the effectiveness of systemic DCZ for DREADD-based chemogenetic manipulations in behavioral and slice electrophysiological applications in rats. We demonstrate that a relatively low dose of DCZ (0.1 mg/kg) supports excitatory DREADD-mediated cFos induction, DREADD-mediated inhibition of a central amygdala-dependent behavior, and DREADD-mediated inhibition of neuronal activity in a slice electrophysiology preparation. In addition, we show that this dose of DCZ does not alter gross locomotor activity or induce a place preference/aversion in control rats without DREADD expression. Together, our findings support the use of systemic DCZ for DREADD-based manipulaations in rats, and provide evidence that DCZ is a superior alternative to CNO.

Connecting the dots between cell populations, whole-brain activity, and behavior.

.Simultaneously manipulating and monitoring both microscopic and macroscopic brain activity in vivo and identifying the linkage to behavior are powerful tools in neuroscience research. These capabilities have been realized with the recent technical advances of optogenetics and its combination with fMRI, here termed “opto-fMRI.” Opto-fMRI allows for targeted brain region-, cell-type-, or projection-specific manipulation and targeted activity measurement to be linked with global brain signaling and behavior. We cover the history, technical advances, applications, and important considerations of opto-fMRI in anesthetized and awake rodents and the future directions of the combined techniques in neuroscience and neuroimaging.

Focused ultrasound excites cortical neurons via mechanosensitive calcium accumulation and ion channel amplification

Ultrasonic neuromodulation has the unique potential to provide non-invasive control of neural activity in deep brain regions with high spatial precision and without chemical or genetic modification. However, the biomolecular and cellular mechanisms by which focused ultrasound excites mammalian neurons have remained unclear, posing significant challenges for the use of this technology in research and potential clinical applications. Here, we show that focused ultrasound excites primary murine cortical neurons in culture through a primarily mechanical mechanism mediated by specific calcium-selective mechanosensitive ion channels. The activation of these channels results in a gradual build-up of calcium, which is amplified by calcium- and voltage-gated channels, generating a burst firing response. Cavitation, temperature changes, large-scale deformation, and synaptic transmission are not required for this excitation to occur. Pharmacological and genetic inhibition of specific ion channels leads to reduced responses to ultrasound, while over-expressing these channels results in stronger ultrasonic stimulation. These findings provide a mechanistic explanation for the effect of ultrasound on neurons to facilitate the further development of ultrasonic neuromodulation and sonogenetics as tools for neuroscience research.

A Versatile Clustered Regularly Interspaced Palindromic Repeats Toolbox to Study Neurological CaV3.2 Channelopathies by Promoter-Mediated Transcription Control.

Precise genome editing in combination with viral delivery systems provides a valuable tool for neuroscience research. Traditionally, the role of genes in neuronal circuits has been addressed by overexpression or knock-out/knock-down systems. However, those techniques do not manipulate the endogenous loci and therefore have limitations. Those constraints include that many genes exhibit extensive alternative splicing, which can be regulated by neuronal activity. This complexity cannot be easily reproduced by overexpression of one protein variant. The CRISPR activation and interference/inhibition systems (CRISPRa/i) directed to promoter sequences can modulate the expression of selected target genes in a highly specific manner. This strategy could be particularly useful for the overexpression of large proteins and for alternatively spliced genes, e.g., for studying large ion channels known to be affected in ion channelopathies in a variety of neurological diseases. Here, we demonstrate the feasibility of a newly developed CRISPRa/i toolbox to manipulate the promoter activity of the Cacna1h gene. Impaired, function of the low-voltage-activated T-Type calcium channel CaV3.2 is involved in genetic/mutational as well as acquired/transcriptional channelopathies that emerge with epileptic seizures. We show CRISPR-induced activation and inhibition of the Cacna1h locus in NS20Y cells and primary cortical neurons, as well as activation in mouse organotypic slice cultures. In future applications, the system offers the intriguing perspective to study functional effects of gain-of-function or loss-of-function variations in the Cacna1h gene in more detail. A better understanding of CaV3.2 channelopathies might result in a major advancement in the pharmacotherapy of CaV3.2 channelopathy diseases.

Gene Expression Profile Changes in the Stimulated Rat Brain Cortex After Repetitive Transcranial Magnetic Stimulation.

Repetitive transcranial magnetic stimulation (rTMS) is gaining popularity as a research tool in neuroscience; however, little is known about its molecular mechanisms of action. The present study aimed to investigate the rTMS-induced transcriptomic changes; we performed microarray messenger RNA, micro RNA, and integrated analyses to explore these molecular events. Eight adult male Sprague-Dawley rats were subjected to a single session of unilateral rTMS at 1 Hz (n = 4) or sham (n = 4). The left hemisphere was stimulated for 20 minutes. To evaluate the cumulative effect of rTMS, eight additional rats were assigned to the 1-Hz (n = 4) or sham (n = 4) rTMS groups. The left hemisphere was stimulated for 5 consecutive days using the same protocol. Microarray analysis revealed differentially expressed genes in the rat cortex after rTMS treatment. The overrepresented gene ontology categories included the positive regulation of axon extension, axonogenesis, intracellular transport, and synaptic plasticity after repeated sessions of rTMS. A single session of rTMS primarily induced changes in the early genes, and several miRNAs were significantly related to the mRNAs. Future studies are required to validate the functional significance of selected genes and refine the therapeutic use of rTMS.

Custom-Built Operant Conditioning Setup for Calcium Imaging and Cognitive Testing in Freely Moving Mice.

Operant chambers are widely used in animal research to study cognition, motivation, and learning processes. Paired with the rapidly developing technologies for brain imaging and manipulations of brain activity, operant conditioning chambers are a powerful tool for neuroscience research. The behavioral testing and imaging setups that are commercially available are often quite costly. Here, we present a custom-built operant chamber that can be constructed in a few days by an unexperienced user with relatively inexpensive, widely available materials. The advantages of our operant setup compared with other open-source and closed-source solutions are its relatively low cost, its support of complex behavioral tasks, its user-friendly setup, and its validated functionality with video imaging of behavior and calcium imaging using the UCLA Miniscope. Using this setup, we replicate our previously published findings showing that mice exposed to social defeat stress in adolescence have inhibitory control impairments in the Go/No-Go task when they reach adulthood. We also present calcium imaging data of medial prefrontal cortex (mPFC) neuronal activity acquired during Go/No-Go testing in freely moving mice and show that neuronal population activity increases from day 1 to day 14 of the task. We propose that our operant chamber is a cheaper alternative to its commercially available counterparts and offers a better balance between versatility and user-friendly setup than other open-source alternatives.

A dual-channel optogenetic stimulator selectively modulates distinct defensive behaviors

SummaryImplantable devices and systems have been emerging as powerful tools for neuroscience research and medical applications. Here we report a wireless, dual-channel optoelectronic system for functional optogenetic interrogation of superior colliculus (SC), a layered structure pertinent to defensive behaviors, in rodents. Specifically, a flexible and injectable probe comprises two thin-film microscale light-emitting diodes (micro-LEDs) at different depths, providing spatially resolved optical illuminations within the tissue. Under remote control, these micro-LEDs interrogate the intermediate layer and the deep layer of the SC (ILSC and DLSC) of the same mice, and deterministically evoke distinct freezing and flight behaviors, respectively. Furthermore, the system allows synchronized optical stimulations in both regions, and we discover that the flight response dominates animals' behaviors in our experiments. In addition, c-Fos immunostaining results further elucidate the functional hierarchy of the SC. These demonstrations provide a viable route to unraveling complex brain structures and functions.

Genetically encoded intrabodies as high-precision tools to visualize and manipulate neuronal function

Basic neuroscience research employs numerous forms of antibodies as key reagents in diverse applications. While the predominant use of antibodies is as immunolabeling reagents, neuroscientists are making increased use of intracellular antibodies or intrabodies. Intrabodies are recombinant antibodies genetically encoded for expression within neurons. These can be used to target various cargo (fluorescent proteins, reporters, enzymes, etc.) to specific molecules and subcellular domains to report on and manipulate neuronal function with high precision. Intrabodies have the advantages inherent in all genetically encoded recombinant antibodies but represent a distinct subclass in that their structure allows for their expression and function within cells. The high precision afforded by the ability to direct their expression to specific cell types, and the selective binding of intrabodies to targets within these allows intrabodies to offer unique advantages for neuroscience research, given the tremendous molecular, cellular and morphological complexity of brain neurons. Intrabodies expressed within neurons have been used for a variety of purposes in basic neuroscience research. Here I provide a general background to intrabodies and their development, and examples of their emerging utility as valuable basic neuroscience research tools.

Nanotechnology Enables Novel Modalities for Neuromodulation.

Neuromodulation is of great importance both as a fundamental neuroscience research tool for analyzing and understanding the brain function, and as a therapeutic avenue for treating brain disorders. Here, an overview of conceptual and technical progress in developing neuromodulation strategies is provided, and it is suggested that recent advances in nanotechnology are enabling novel neuromodulation modalities with less invasiveness, improved biointerfaces, deeper penetration, and higher spatiotemporal precision. The use of nanotechnology and the employment of versatile nanomaterials and nanoscale devices with tailored physical properties have led to considerable research progress. To conclude, an outlook discussing current challenges and future directions for next-generation neuromodulation modalities is presented.

Natural and Artificial Intelligence: A brief introduction to the interplay between AI and neuroscience research

Neuroscience and artificial intelligence (AI) share a long history of collaboration. Advances in neuroscience, alongside huge leaps in computer processing power over the last few decades, have given rise to a new generation of in silico neural networks inspired by the architecture of the brain. These AI systems are now capable of many of the advanced perceptual and cognitive abilities of biological systems, including object recognition and decision making. Moreover, AI is now increasingly being employed as a tool for neuroscience research and is transforming our understanding of brain functions. In particular, deep learning has been used to model how convolutional layers and recurrent connections in the brain’s cerebral cortex control important functions, including visual processing, memory, and motor control. Excitingly, the use of neuroscience-inspired AI also holds great promise for understanding how changes in brain networks result in psychopathologies, and could even be utilized in treatment regimes. Here we discuss recent advancements in four areas in which the relationship between neuroscience and AI has led to major advancements in the field; (1) AI models of working memory, (2) AI visual processing, (3) AI analysis of big neuroscience datasets, and (4) computational psychiatry.

Implantable Optrode Array for Optogenetic Modulation and Electrical Neural Recording.

During the last decade, optogenetics has become an essential tool for neuroscience research due to its unrivaled feature of cell-type-specific neuromodulation. There have been several technological advances in light delivery devices. Among them, the combination of optogenetics and electrophysiology provides an opportunity for facilitating optogenetic approaches. In this study, a novel design of an optrode array was proposed for realizing optical modulation and electrophysiological recording. A 4 × 4 optrode array and five-channel recording electrodes were assembled as a disposable part, while a reusable part comprised an LED (light-emitting diode) source and a power line. After the characterization of the intensity of the light delivered at the fiber tips, in vivo animal experiment was performed with transgenic mice expressing channelrhodopsin, showing the effectiveness of optical activation and neural recording.

Shedding light on neuroscience: Two decades of functional near-infrared spectroscopy applications and advances from a bibliometric perspective.

Functional near-infrared spectroscopy (fNIRS) is a noninvasive optical brain-imaging technique that detects changes in hemoglobin concentration in the cerebral cortex. fNIRS devices are safe, silent, portable, robust against motion artifacts, and have good temporal resolution. fNIRS is reliable and trustworthy, as well as an alternative and a complement to other brain-imaging modalities, such as electroencephalography or functional magnetic resonance imaging. Given these advantages, fNIRS has become a well-established tool for neuroscience research, used not only for healthy cortical activity but also as a biomarker during clinical assessment in individuals with schizophrenia, major depressive disorder, bipolar disease, epilepsy, Alzheimer's disease, vascular dementia, and cancer screening. Owing to its wide applicability, studies on fNIRS have increased exponentially over the last two decades. In this study, scientific publications indexed in the Web of Science databases were collected and a bibliometric-type methodology was developed. For this purpose, a comprehensive science mapping analysis, including top-ranked authors, journals, institutions, countries, and co-occurring keywords network, was conducted. From a total of 2310 eligible documents, 6028 authors and 531 journals published fNIRS-related papers, Fallgatter published the highest number of articles and was the most cited author. University of Tübingen in Germany has produced the most trending papers since 2000. USA was the most prolific country with the most active institutions, followed by China, Japan, Germany, and South Korea. The results also revealed global trends in emerging areas of research, such as neurodevelopment, aging, and cognitive and emotional assessment.

Inhibition of Na+/H+ exchanger modulates microglial activation and scar formation following microelectrode implantation

Objective. Intracortical microelectrodes are an important tool for neuroscience research and have great potential for clinical use. However, the use of microelectrode arrays to treat neurological disorders and control prosthetics is limited by biological challenges such as glial scarring, which can impair chronic recording performance. Microglia activation is an early and prominent contributor to glial scarring. After insertion of an intracortical microelectrode, nearby microglia transition into a state of activation, migrate, and encapsulate the device. Na+/H+ exchanger isoform-1 (NHE-1) is involved in various microglial functions, including their polarity and motility, and has been implicated in pro-inflammatory responses to tissue injury. HOE-642 (cariporide) is an inhibitor of NHE-1 and has been shown to depress microglial activation and inflammatory response in brain injury models. Approach. In this study, the effects of HOE-642 treatment on microglial interactions to intracortical microelectrodes was evaluated using two-photon microscopy in vivo. Main results. The rate at which microglia processes and soma migrate in response to electrode implantation was unaffected by HOE-642 administration. However, HOE-642 administration effectively reduced the radius of microglia activation at 72 h post-implantation from 222.2 µm to 177.9 µm. Furthermore, treatment with HOE-642 significantly reduced microglial encapsulation of implanted devices at 5 h post-insertion from 50.7 ± 6.0% to 8.9 ± 6.1%, which suggests an NHE-1-specific mechanism mediating microglia reactivity and gliosis during implantation injury. Significance. This study implicates NHE-1 as a potential target of interest in microglial reactivity and HOE-642 as a potential treatment to attenuate the glial response and scar formation around implanted intracortical microelectrodes.

Hematology and Culture Assessment of Cranially Implanted Rhesus Macaques (Macaca mulatta).

The use of percutaneous cranial implants in rhesus macaques (Macaca mulatta) has long been a valuable tool for neuroscience research. However, when treating and assessing these animals, veterinarians are required to make assumptions about diagnostic results due to a lack of research into how these implants affect physiology. Microbial cultures of cranial implant sites show an abundance of colonizing bacteria, but whether these microbes affect animal health and wellbeing is poorly understood. In addition, microbial antibiotic resistance can present significant health concerns for both the animals and the researchers. To help elucidate the relationship between percutaneous cranial implants and blood parameters, complete blood cell counts and serum chemistry results were assessed on 57 nonhuman primates at our institution from September 2001 to March 2017. Generalized estimating equations were used to compare the results before and after an animal's first implant surgery. This modelling showed that cranial implants were a significant predictor of alterations in the number of neutrophils, lymphocytes, and red blood cells, and in the concentration of hemoglobin, alkaline phosphatase, creatinine, calcium, phos- phorus, total protein, albumin, and globulin. Anaerobic and aerobic bacterial cultures were performed to identify bacteria associated with cranial implants. Staphylococcus spp., Streptococcus spp., and Corynebacterium spp. comprised the majority of the aerobic bacterial isolates, while Fusobacterium spp., Peptostreptococcus spp. and Bacterioides fragilis comprised the majority of anaerobic bacterial isolates. Using a Pearson r correlation for statistical analysis, we assessed whether any of these bacterial isolates developed antibiotic resistances over time. Cefazolin, the most frequently used antibiotic in monkeys in this study, was the only antimicrobial out of 41 agents tested to which bacteria developed resistance over time. These results indicate that percutaneous implants are associated with a generalized inflammatory state, multiple bacterial species are present at the implant site, and these bacteria may contribute to the inflammatory response.

Multimodal Functional Analysis Platform: 1. Ultrathin Fluorescence Endoscope Imaging System Enables Flexible Functional Brain Imaging.

To elucidate the expression mechanisms of brain functions, we have developed an ultrathin fluorescence endoscope imaging system (U-FEIS) that can image cells in the brain at any depth while minimizing the invasion. The endoscope part of U-FEIS consists of a GRIN lens and a 10,000-pixel image fiber with a diameter of 450μm. The specialized microscope of U-FEIS is within 30cm square and includes lenses and optical filters optimized for the endoscope. Using U-FEIS, we successfully visualized neurons expressing GFP with single-cell resolution and recorded the multineuronal activities in vitro and in vivo. U-FEIS can also perform imaging and optical stimulation simultaneously. Therefore, U-FEIS should be a powerful optical tool in neuroscience research.

Discovery of novel peptide neurotoxins from sea anemone species.

As primitive metazoa, sea anemones are rich in various bioactive peptide neurotoxins. These peptides have been applied to neuroscience research tools or directly developed as marine drugs. To date, more than 1100 species of sea anemones have been reported, but only 5% of the species have been used to isolate and identify sea anemone peptide neurotoxins. There is an urgent need for more systematic discovery and study of peptide neurotoxins in sea anemones. In this review, we have gathered the currently available methods from crude venom purification and gene cloning to venom multiomics, employing these techniques for discovering novel sea anemone peptide neurotoxins. In addition, the three-dimensional structures and targets of sea anemone peptide neurotoxins are summarized. Therefore, the purpose of this review is to provide a reference for the discovery, development, and utilization of sea anemone peptide neurotoxins.

Incorporating Neuroscience Data into Agent-Based Simulation Models of Buyer Behavior

Purpose: The article aims to analyze the possibility of using various cognitive neuroscience techniques when building the agent model of buyer behavior and propose an experimental procedure for obtaining qualitative data based on the triangulation of methods. Design/Methodology/Approach: The proposed approach combines agent-based simulation with cognitive neuroscience techniques at the stage of designing the characteristics and behavior rules of agents-consumers. Findings: The consumer’s purchasing behavior is determined by the compilation of the influence of environmental factors and marketing stimuli as well as by his personality traits. Due to the necessity to consider all these elements when mapping the consumer-agent characteristics and decision rules, traditional methods of data collection may not be sufficient. In such a situation, cognitive neuroscience techniques can become a source of additional information, allowing to take into account the influence of emotions or cognitive abilities on one’s decisions. To make it possible, it is necessary to conduct experiments with the use of neuroscience research tools (e.g., EEG, GSR, HR etc.) aimed at detecting emotional and cognitive states during exposure to an advertisement of a specific product. The neurophysiological data collected during the experiments allow for a more accurate estimation of the qualitative parameters describing consumer behavior rules. Practical Implications: The proposed concept allows for a more accurate representation of agents-consumers’ features and decision rules. Consequently, the agent-based model more reliably reflects reality, and thus the results obtained during model simulation are more valuable and can be the basis for formulating marketing plans. Originality/Value: The proposed approach enriches the methodology of data collection and estimation of qualitative parameters in building agent models of buyer behavior.