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<em>In Vivo</em> Two-photon Imaging of Cortical Neurons in Neonatal Mice
摘要： Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging methods exist for examining the brain tissue of live newborn mammals. Herein we summarize a protocol for imaging individual cortical neurons in living neonatal mice. This protocol includes the following two methodologies: (1) the Supernova system for sparse and bright labeling of cortical neurons in the developing brain, and (2) a surgical procedure for the fragile neonatal skull. This protocol allows the observation of temporal changes of individual cortical neurites during neonatal stages with a high signal-to-noise ratio. Labeled cell-specific gene silencing and knockout can also be achieved by combining the Supernova with RNA interference and CRISPR/Cas9 gene editing systems. This protocol can, thus, be used for analyzing the developmental dynamics of cortical neurons, molecular mechanisms that control the neuronal dynamics, and changes in neuronal dynamics in disease models.
关键词： Neuroscience,in vivo imaging,single-cell labeling,mouse,cerebral cortex,Newborn,two-photon
更新于2025-09-23 15:21:01
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A Protocol for Transcranial Photobiomodulation Therapy in Mice
摘要： Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide range of neurological and psychiatric disorders, and memory enhancement in age-related cognitive decline and neurodegenerative diseases. We describe a laboratory protocol for transcranial photobiomodulation therapy (PBMT) in mice. Aged BALB/c mice (18 months old) are treated with a 660 nm laser transcranially, once daily for 2 weeks. Laser transmittance data shows that approximately 1% of the incident red light on the scalp reaches a 1 mm depth from the cortical surface, penetrating the dorsal hippocampus. Treatment outcomes are assessed by two methods: a Barnes maze test, which is a hippocampus-dependent spatial learning and memory task evaluation, and measuring hippocampal ATP levels, which is used as a bioenergetics index. The results from the Barnes task show an enhancement of the spatial memory in laser-treated aged mice when compared with age-matched controls. Biochemical analysis after laser treatment indicates increased hippocampal ATP levels. We postulate that the enhancement of memory performance is potentially due to an improvement in hippocampal energy metabolism induced by the red laser treatment. The observations in mice could be extended to other animal models since this protocol could potentially be adapted to other species frequently used in translational neuroscience, such as rabbit, cat, dog, or monkey. Transcranial photobiomodulation is a safe and cost-effective modality which may be a promising therapeutic approach in age-related cognitive impairment.
关键词： low-level laser therapy,optical properties,red light,hippocampus,mouse,aging,Transcranial photobiomodulation,learning,memory,Issue 141,Neuroscience
更新于2025-09-23 15:21:01
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Glymphatic clearance of simulated silicon dispersion in mouse brain analyzed by laser induced breakdown spectroscopy
摘要： Silicon-based devices, such as neural probes, are increasingly used as electrodes for receiving electrical signals from neural tissue. Neural probes used chronically have been known to induce inflammation and elicit an immune response. The current study detects and evaluates silicon dispersion from a concentrated source in the mouse brain using laser induced breakdown spectroscopy. Element lines for Si (I) were found at the injection site at approximately 288 nm at 3hr post-implantation, even with tissue perfusion, indicating possible infusion into neural tissue. At 24hr and 1-week post-implantation, no silicon lines were found, indicating clearance. An isolated immune response was found by CD68 macrophage response at 24hr post injection. Future studies should measure chronic silicon exposure to determine if the inflammatory response is proportional to silicon administration. The present type of protocol, coupling laser induced breakdown spectroscopy, neuroimaging, histology, immunohistochemistry, and determination of clearance could be used to investigate the glymphatic system and different tissue states such as in disease (e.g. Alzheimer's).
关键词： Inflammation,Silicon,Neuroscience,Biophysics,Physics methods,Probe,Toxicology,Physics,Optics,Immune response,Electrode
更新于2025-09-23 15:19:57
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Optical Monitoring of Living Nerve Terminal Labeling in Hair Follicle Lanceolate Endings of the <em>Ex Vivo</em> Mouse Ear Skin
摘要： A novel dissection and recording technique is described for optical monitoring staining and de-staining of lanceolate terminals surrounding hair follicles in the skin of the mouse pinna. The preparation is simple and relatively fast, reliably yielding extensive regions of multiple labeled units of living nerve terminals to study uptake and release of styryl pyridinium dyes extensively used in studies of vesicle recycling. Subdividing the preparations before labeling allows test vs. control comparisons in the same ear from a single individual. Helpful tips are given for improving the quality of the preparation, the labeling and the imaging parameters. This new system is suitable for assaying pharmacologically and mechanically-induced uptake and release of these vital dyes in lanceolate terminals in both wild-type and genetically modified animals. Examples of modulatory influences on labeling intensity are given.
关键词： ear,Issue 110,Neuroscience,mechanosensation,hair follicle,electrophysiology,mouse,Lanceolate ending,skin
更新于2025-09-19 17:15:36
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[IEEE 2018 International Joint Conference on Neural Networks (IJCNN) - Rio de Janeiro (2018.7.8-2018.7.13)] 2018 International Joint Conference on Neural Networks (IJCNN) - Investigation into Sub-Receptive Fields of Retinal Ganglion Cells with Natural Images
摘要： Determining the receptive field of a retinal ganglion cell is critically important when formulating a computational model that maps the relationship between the stimulus and response. This process is traditionally undertaken using reverse correlation to estimate the receptive field. By stimulating the retina with artificial stimuli, such as alternating checkerboards, bars or gratings and recording the neural response it is possible to estimate the cell’s receptive field by analysing the stimuli that produced the response. Artificial stimuli such as white noise is known to not stimulate the full range of the cell’s responses. By using natural image stimuli, it is possible to estimate the receptive field and obtain a resulting model that more accurately mimics the cells’ responses to natural stimuli. This paper extends on previous work to seek further improvements in estimating a ganglion cell’s receptive field by considering that the receptive field can be divided into subunits. It is thought that these subunits may relate to receptive fields which are associated with bipolar retinal cells. The findings of this preliminary study show that by using subunits to define the receptive field we achieve a significant improvement over existing approaches when deriving computational models of the cell’s response.
关键词： computational modelling,visual neuroscience,receptive field,retinal ganglion cell
更新于2025-09-19 17:15:36
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Conducting Hyperscanning Experiments with Functional Near-Infrared Spectroscopy
摘要： Concurrent brain recordings of two or more interacting persons, an approach termed hyperscanning, are gaining increasing importance for our understanding of the neurobiological underpinnings of social interactions, and possibly interpersonal relationships. Functional near-infrared spectroscopy (fNIRS) is well suited for conducting hyperscanning experiments because it measures local hemodynamic effects with a high sampling rate and, importantly, it can be applied in natural settings, not requiring strict motion restrictions. In this article, we present a protocol for conducting fNIRS hyperscanning experiments with parent-child dyads and for analyzing brain-to-brain synchrony. Furthermore, we discuss critical issues and future directions, regarding the experimental design, spatial registration of the fNIRS channels, physiological influences and data analysis methods. The described protocol is not specific to parent-child dyads, but can be applied to a variety of different dyadic constellations, such as adult strangers, romantic partners or siblings. To conclude, fNIRS hyperscanning has the potential to yield new insights into the dynamics of the ongoing social interaction, which possibly go beyond what can be studied by examining the activities of individual brains.
关键词： neuroscience,brain-to-brain synchrony,cooperation,fNIRS,parent-child interaction,hyperscanning,functional near-infrared spectroscopy
更新于2025-09-19 17:15:36
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[ACM Press the 2018 ACM International Symposium - Singapore, Singapore (2018.10.08-2018.10.12)] Proceedings of the 2018 ACM International Symposium on Wearable Computers - ISWC '18 - Seeing into the brain of an actor with mocap and fNIRS
摘要： This paper introduces the idea of using wearable, multi-modal body and brain sensing, in a theatrical setting, for neuroscientific research. Wearable motion capture suits are used to track the body movements of two actors while they enact a sequence of scenes together. One actor additionally wears a functional near-infrared spectroscopy (fNIRS)-based headgear to record the activation patterns on his prefrontal cortex. Repetitions in the movement data are then used to automatically segment the fNIRS data for further analysis. This exploration reveals that the semi-structured and repeatable nature of theatre can provide a useful laboratory for neuroscience, and that wearable sensing is a promising method to achieve this. This is important because it points to a new way of researching the brain in a more natural, and social, environment than traditional lab-based methods.
关键词： Wearable,fNIRS,theatre,mocap,neuroscience
更新于2025-09-11 14:15:04
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Expanding the optogenetics toolkit
摘要： Optogenetics has been instrumental for the dissection of functional neural circuits. As this branch of neuroscience flourishes, and as experimental designs become increasingly sophisticated, the need for new opsins—light-activated proteins that can be used to excite or inhibit specific neuronal populations—also grows. Previous efforts have relied on either the discovery of naturally occurring opsin variants with unique features through genome mining, or the engineering of opsins to achieve certain desired properties. Now, researchers at the Janelia Research Campus of the Howard Hughes Medical Institute, led by Joshua Dudman and Alla Karpova, have developed a new protein-engineering approach that could in principle double the number of viable tools for optogenetics. Taking inspiration from evolution, the researchers wondered whether topological inversion—flipping a protein inside-out within the membrane—could generate opsin variants with new properties. They designed a leader sequence containing the signal peptide and the transmembrane domain of neurexin 1B-δ, and they packed in a few more tweaks to further support its desired orientation. They then fused the leader sequence and a well-characterized opsin, ChR2 E123T/T159C. This effectively flipped the opsin within the membrane. Strikingly, the opsin was transformed from an optogenetic activator to a potent inhibitor that functions as a light-activated, nonselective cation pump. They dubbed this new variant ‘full-length inversion of ChR’ (FLInChR). FLInChR suppressed the activity of SNr GABAergic neurons in brain slices. In vivo, FLInChR was able to inhibit SNr projection neurons and modulate animal behaviors. Notably, the extent of inhibition elicited by FLInChR was on par with that observed with ArchT, a commonly used optogenetic inhibitor. It is worth noting that nonselective cation pumping has not been previously achieved through directed molecular engineering. This topological engineering approach could help to create new optogenetic tools for neuroscience research.
关键词： neuroscience,opsins,optogenetics,protein engineering,topological inversion
更新于2025-09-10 09:29:36
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Neuroergonomics || Functional Near-Infrared Spectroscopy
摘要： The mission of social psychology is to understand “how the thoughts, feelings, and behaviour of individuals are influenced by the actual, imagined, or implied presence of others.” The mission of social neuroscience is to understand the neural and broader biological underpinnings of social psychological phenomena. Like every scientific discipline, the success of social neuroscience rests on the effectiveness of its methods. Recent years have witnessed a proliferation of methods that hold promise for advancing social neuroscience. One such method is continuous-wave functional near-infrared spectroscopy (fNIRS), a functional neuroimaging technique that can be used to measure brain activity noninvasively. In this chapter we demonstrate the utility of fNIRS for social neuroscience in identifying well-established patterns of prefrontal activity when people make self- and other-referential judgments.
关键词： social neuroscience,self-referential judgments,prefrontal activity,functional near-infrared spectroscopy,fNIRS
更新于2025-09-10 09:29:36
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An optimized and automated approach to quantifying channelrhodopsin photocurrent kinetics
摘要： Channelrhodopsins are light-activated ion channels that enable targetable activation or inhibition of excitable cells with light. Ion conductance can generally be described by a four step photocycle, which includes two open and two closed states. While a complete understanding of channelrhodopsin function cannot be understood in the absence of kinetic modeling, model fitting requires manual fitting, which is laborious and technically complicated for non-experts. To enhance analysis of photocurrent data, this manuscript describes a fitting program where electrophysiology data can be automatically and quantitatively analyzed. Significant improvement in this program when compare to our previous version includes 1) the ability to automatically find the experiment start time using the derivative of the current signal, 2) utilizing the Object Oriented Programing (OPP) paradigm which is significantly more reliable if the code is used by people with little to no programming experience and 3) the distribution of the code is simplified to sharing a single MATLAB file, including rigorous comments throughout. To demonstrate the utility of this program, we show automated fitting of photocurrents from two member proteins: channelrhodopsin-2 and a chimera between channelrhodopsin-1 and channelrhodopsin-2 (C1C2).
关键词： neuroscience,channelrhodopsin,optogenetics,kinetic modeling,electrophysiology
更新于2025-09-10 09:29:36