この作品は、国立科学財団（BES-0447713）からとエシロール、国際からの助成金からCAREER賞によって部分的にサポートされていました。

<ol>
	<li>Leigh, R. J., Zee, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Neurology of Eye Movements. , (2006).</li><li>Pruehsner, W. R., Enderle, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+operating+version+of+the+Eye+Tracker,+a+system+to+measure+saccadic+eye+movements.">The operating version of the Eye Tracker, a system to measure saccadic eye movements.</a> Biomed Sci Instrum. 38, 113-118 (2002).</li><li>Pruehsner, W. R., Liebler, C. M., Rodriguez-Campos, F., Enderle, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Eye+Tracker+System--a+system+to+measure+and+record+saccadic+eye+movements.">The Eye Tracker System--a system to measure and record saccadic eye movements.</a> Biomed Sci Instrum. 39, 208-213 (2003).</li><li>Rufa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-based+eye+tracking:+our+experience+with+Advanced+Stimuli+Design+for+Eye+Tracking+software.">Video-based eye tracking: our experience with Advanced Stimuli Design for Eye Tracking software.</a> Ann N Y Acad Sci. 1039, 575-579 (2005).</li><li>Cornelissen, F. W., Peters, E. M., Palmer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Eyelink+Toolbox:+eye+tracking+with+MATLAB+and+the+Psychophysics+Toolbox.">The Eyelink Toolbox: eye tracking with MATLAB and the Psychophysics Toolbox.</a> Behav Res Methods Instrum Comput. 34, 613-617 (2002).</li><li>Han, S. J., Guo, Y., Granger-Donetti, B., Vicci, V. R., Alvarez, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+heterophoria+and+phoria+adaptation+using+an+automated+objective+system+compared+to+clinical+methods.">Quantification of heterophoria and phoria adaptation using an automated objective system compared to clinical methods.</a> Ophthalmic Physiol Opt. 30, 95-107 (2010).</li><li>Kim, E. H., Granger-Donetti, B., Vicci, V. R., Alvarez, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Relationship+between+Phoria+and+the+Ratio+of+Convergence+Peak+Velocity+to+Divergence+Peak+Velocity.">The Relationship between Phoria and the Ratio of Convergence Peak Velocity to Divergence Peak Velocity.</a> Invest Ophthalmol Vis Sci. , (2010).</li><li>Lee, Y. Y., Granger-Donetti, B., Chang, C., Alvarez, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+convergence+induced+changes+in+phoria+and+divergence+dynamics.">Sustained convergence induced changes in phoria and divergence dynamics.</a> Vision Res. 49, 2960-2972 (2009).</li><li>Maruta, J., Suh, M., Niogi, S. N., Mukherjee, P., Ghajar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+tracking+synchronization+as+a+metric+for+concussion+screening.">Visual tracking synchronization as a metric for concussion screening.</a> J Head Trauma Rehabil. 25, 293-305 (2010).</li><li>Osanai, R., Kinoshita, M., Hirose, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+movement+disorders+in+myotonic+dystrophy+type+1.">Eye movement disorders in myotonic dystrophy type 1.</a> Acta Otolaryngol Suppl. , 78-84 (2007).</li><li>Kaufman, L. D., Pratt, J., Levine, B., Black, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antisaccades:+a+probe+into+the+dorsolateral+prefrontal+cortex+in+Alzheimer's+disease.+A+critical+review.">Antisaccades: a probe into the dorsolateral prefrontal cortex in Alzheimer's disease. A critical review.</a> J Alzheimers Dis. 19, 781-793 (2010).</li><li>Hannula, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+Eye+Movement+Monitoring+to+Examine+Item+and+Relational+Memory+in+Schizophrenia.">Use of Eye Movement Monitoring to Examine Item and Relational Memory in Schizophrenia.</a> Biol Psychiatry. , (2010).</li><li>Schubert, M. C., Zee, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saccade+and+vestibular+ocular+motor+adaptation.">Saccade and vestibular ocular motor adaptation.</a> Restor Neurol Neurosci. 28, 9-18 (2010).</li><li>Noudoost, B., Chang, M. H., Steinmetz, N. A., Moore, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Top-down+control+of+visual+attention.">Top-down control of visual attention.</a> Curr Opin Neurobiol. 20, 183-190 (2010).</li><li>Herwig, A., Beisert, M., Schneider, W. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+spatial+interaction+of+visual+working+memory+and+attention:+evidence+for+a+global+effect+from+memory-guided+saccades.">On the spatial interaction of visual working memory and attention: evidence for a global effect from memory-guided saccades.</a> J Vis. 10, (2010).</li><li>McDowell, J. E., Dyckman, K. A., Austin, B. P., Clementz, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurophysiology+and+neuroanatomy+of+reflexive+and+volitional+saccades:+evidence+from+studies+of+humans.">Neurophysiology and neuroanatomy of reflexive and volitional saccades: evidence from studies of humans.</a> Brain Cogn. 68, 255-270 (2008).</li></ol>

利害の衝突が宣言されていない。

重要なステップ： 眼球運動のモニターが適切に被写体に調整する必要があります。例えば、眼球運動記録モニターは、範囲内で動作し、被験者に調節されなければならない。被検眼の動きの範囲を超えた場合、システムが飽和する。飽和時には、眼球運動信号が有効ではありません。キャリブレーションは、また眼球運動記録において非常に重要です。全眼球運動モニタ?...

A correction was made to <a href="http://www.jove.com/details.php?id=2530">VisualEyes: A Modular Software System for Oculomotor Experimentation</a>. There was an error in the authors, Eun H. Kim and Tara L. Alvarez, names. The author's names have been corrected to: Eun H. Kim and Tara L. Alvarez instead of: Eun Kim and Tara Alvarez

Erratum: VisualEyes: A Modular Software System for Oculomotor Experimentation

visualeyes: a modular software system for oculomotor experimentation

眼球運動の研究では、脳は、両方の正常な機能不全、脳内の視覚的な情報を取得する方法の理解を形成する強力な基盤を提供してきた。1しかし、プラットフォームの開発を刺激するために、店舗目の動きが実質的なプログラミング、時間とコストを必要とすることができます。多くのシステムでは、実験的なニーズの多様のための多数の刺激をプログラムするための柔軟性を提供していません。しかし、VisualEyesシステムは、オペレータが任意の背景と前景の刺激、プログラムタンデムまたは目の動きに対向するための1つまたは2つの画面を選択し、独立して、左右の眼を刺激することができ、柔軟なアーキテクチャを有する。このシステムは、大幅に動眼研究を行うために必要なプログラミングの開発時間を減らすことができる。刺激のアレイを生成するために、眼球運動の応答を取得するための1）眼球運動記録装置のLabViewで書か2）VisualEyesソフトウェア：VisualEyesシステムは3つの部分に説明されるテキストフ​​ァイルおよび3）オフラインデータ分析としてストア応答。輪部追跡システム、強膜サーチコイル、またはビデオ画像システム：眼球運動のような器具のいくつかの種類によって記録することができる。そのようなサッカードの手順などの典型的な眼球運動刺激は、vergentランプと対応する応答とのvergentのステップが表示されます。このビデオレポートでは、多数の視覚刺激と健康だけでなく、臨床集団を研究するために、基本的な科学者や臨床医によって利用することができ、レコード目の動きを作成するためのシステムの柔軟性を実証する。

Watch this Scientific Journal Video about VisualEyes: A Modular Software System for Oculomotor Experimentation at JoVE.com

VisualEyes: A Modular Software System for Oculomotor Experimentation

神経制御および認知プロセスは、眼球運動を介して研究することができる。 VisualEyesソフトウェアは、オペレータが、独立して、単純な、カスタムスクリプト言語を使用して2つのコンピュータ画面上に刺激をプログラムすることができます。システムは、タンデム眼球運動（サッカードと円滑追跡）、または反対の眼の動き（輻輳）、または任意の組み合わせを刺激することができる。

VisualEyes：眼球運動実験のためのモジュラ式のソフトウェアシステム

Eye movement studies have provided a strong foundation forming an understanding of how the brain acquires visual information in both the normal and ...

visualeyes-a-modular-software-system-for-oculomotor-experimentation

Research

JoVE Journal

Neuroscience

12.6K ビュー.  New Jersey Institute of Technology. 神経制御および認知プロセスは、眼球運動を介して研究することができる。 VisualEyesソフトウェアは、オペレータが、独立して、単純な、カスタムスクリプト言語を使用して2つのコンピュータ画面上に刺激をプログラムすることができます。システムは、タンデム眼球運動（サッカードと円滑追跡）、または反対の眼の動き（輻輳）、または任意の組み?...

ビデオ: VisualEyes: A Modular Software System for Oculomotor Experimentation

A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously published phenotype assessments in several disease models, including spinocerebellar ataxias, Huntington s disease and spinobulbar muscular atrophy. Measures include hind limb clasping, ledge test, gait and kyphosis. Each measure is recorded on a scale of 0-3, with a combined total of 0-12 for all four measures. The results effectively discriminate between affected and non-affected individuals, while also quantifying the temporal progression of neurodegenerative disease phenotypes. Measures may be analyzed individually or combined into a composite phenotype score for greater statistical power. The ideal combination of the four described measures will depend upon the disorder in question. We present an example of the protocol used to assess disease severity in a transgenic mouse model of spinocerebellar ataxia type 7 (SCA7).
Albert R. La Spada and Gwenn A. Garden contributed to this manuscript equally.

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebellar ataxia. Measures include hind limb clasping, ledge test, gait and kyphosis. This protocol effectively discriminates between affected and non-affected individuals, and detects the progression of affected individuals over time.

小脳性運動失調のマウスモデルを評価するための単純な複合表現型のスコアリングシステム

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously ...

神経科学

Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings with a force transducer to monitor the strength of contraction. In addition, we demonstrate how to visually monitor peristaltic activity, which can be used as a bioassay for various peptides, biogenic amines and neurotransmitters. This preparation is amenable to student laboratories in physiology and for demonstrating pharmacological concepts to students. This preparation has been in use for over 100 years, and it still offers much as a model for investigating the generation and regulation of peristaltic rhythms and for describing the mechanisms underlying their modulation. The pharmacological assays and receptor sub-typing that were started over 50 years ago on the hindgut still contribute to research today. This robust preparation is well suited to training students in physiology and pharmacology.

In this report we demonstrate techniques that can be used to investigate the biology of the crayfish hindgut. We show how to dissect a crayfish abdomen and study the associated anatomy, physiology and modulation of activity. The peristaltic activity and strength of contractions are measured using a force transducer.

ザリガニ後腸との生理学的実験：学生実験室実習

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings ...

VisioTracker, an Innovative Automated Approach to Oculomotor Analysis

Investigations into the visual system development and function necessitate quantifiable behavioral models of visual performance that are easy to elicit, robust, and simple to manipulate. A suitable model has been found in the optokinetic response (OKR), a reflexive behavior present in all vertebrates due to its high selection value. The OKR involves slow stimulus-following movements of eyes alternated with rapid resetting saccades. The measurement of this behavior is easily carried out in zebrafish larvae, due to its early and stable onset (fully developed after 96 hours post fertilization (hpf)), and benefitting from the thorough knowledge about zebrafish genetics, for decades one of the favored model organisms in this field. Meanwhile the analysis of similar mechanisms in adult fish has gained importance, particularly for pharmacological and toxicological applications.
Here we describe VisioTracker, a fully automated, high-throughput system for quantitative analysis of visual performance. The system is based on research carried out in the group of Prof. Stephan Neuhauss and was re-designed by TSE Systems. It consists of an immobilizing device for small fish monitored by a high-quality video camera equipped with a high-resolution zoom lens. The fish container is surrounded by a drum screen, upon which computer-generated stimulus patterns can be projected. Eye movements are recorded and automatically analyzed by the VisioTracker software package in real time.
Data analysis enables immediate recognition of parameters such as slow and fast phase duration, movement cycle frequency, slow-phase gain, visual acuity, and contrast sensitivity.
Typical results allow for example the rapid identification of visual system mutants that show no apparent alteration in wild type morphology, or the determination of quantitative effects of pharmacological or toxic and mutagenic agents on visual system performance.

The VisioTracker is an automated system for the quantitative analysis of visual performance of larval and small adult fish based on the recording of eye movements. It features full control over visual stimulus properties and real-time analysis, enabling high-throughput research in fields such as visual system development and function, pharmacology, neural circuit studies and sensorimotor integration.

VisioTracker、眼球運動解析への革新的な自動化されたアプローチ

Investigations into the visual system development and function necessitate quantifiable behavioral models of visual performance that are easy to elicit, ...

A Lightweight, Headphones-based System for Manipulating Auditory Feedback in Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and sensorimotor plasticity1. Consistent sensory perturbations result in compensatory changes in motor output, reflecting changes in feedforward motor control that reduce the experienced feedback error. By quantifying how different sensory feedback errors affect human behavior, prior studies have explored how visual signals are used to recalibrate arm movements2,3 and auditory feedback is used to modify speech production4-7. The strength of this approach rests on the ability to mimic naturalistic errors in behavior, allowing the experimenter to observe how experienced errors in production are used to recalibrate motor output.
Songbirds provide an excellent animal model for investigating the neural basis of sensorimotor control and plasticity8,9. The songbird brain provides a well-defined circuit in which the areas necessary for song learning are spatially separated from those required for song production, and neural recording and lesion studies have made significant advances in understanding how different brain areas contribute to vocal behavior9-12. However, the lack of a naturalistic error-correction paradigm - in which a known acoustic parameter is perturbed by the experimenter and then corrected by the songbird - has made it difficult to understand the computations underlying vocal learning or how different elements of the neural circuit contribute to the correction of vocal errors13.
The technique described here gives the experimenter precise control over auditory feedback errors in singing birds, allowing the introduction of arbitrary sensory errors that can be used to drive vocal learning. Online sound-processing equipment is used to introduce a known perturbation to the acoustics of song, and a miniaturized headphones apparatus is used to replace a songbird's natural auditory feedback with the perturbed signal in real time. We have used this paradigm to perturb the fundamental frequency (pitch) of auditory feedback in adult songbirds, providing the first demonstration that adult birds maintain vocal performance using error correction14. The present protocol can be used to implement a wide range of sensory feedback perturbations (including but not limited to pitch shifts) to investigate the computational and neurophysiological basis of vocal learning.

We describe the design and assembly of miniaturized headphones suitable for replacing a songbird&#x2019;s natural auditory feedback with a manipulated acoustic signal. Online sound processing hardware is used to manipulate song output, introduce real-time errors in auditory feedback via the headphones, and drive vocal motor learning.

鳴禽類には聴覚フィードバックを操作するための軽量、ヘッドフォンベースのシステム

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and ...

A Caenorhabditis elegans Model System for Amylopathy Study

Amylopathy is a term that describes abnormal synthesis and accumulation of amyloid beta (A&beta;) in tissues with time. A&beta; is a hallmark of Alzheimer's disease (AD) and is found in Lewy body dementia, inclusion body myositis and cerebral amyloid angiopathy 1-4. Amylopathies progressively develop with time. For this reason simple organisms with short lifespans may help to elucidate molecular aspects of these conditions. Here, we describe experimental protocols to study A&beta;-mediated neurodegeneration using the worm Caenorhabditis elegans. Thus, we construct transgenic worms by injecting DNA encoding human A&beta;42 into the syncytial gonads of adult hermaphrodites. Transformant lines are stabilized by a mutagenesis-induced integration. Nematodes are age synchronized by collecting and seeding their eggs. The function of neurons expressing A&beta;42 is tested in opportune behavioral assays (chemotaxis assays). Primary neuronal cultures obtained from embryos are used to complement behavioral data and to test the neuroprotective effects of anti-apoptotic compounds.

A Caenorhabditis elegans Model System for Amylopathy Study

We describe methods to study aspects of amylopathies in the worm C. elegans. We show how to construct worms expressing human A&beta;42 in neurons and how to test their function in behavioral assays. We further show how to obtain primary neuronal cultures that can be used for pharmacological testing.

A<em&gt;線虫（Caenorhabditis elegans）</emAmylopathyの研究のための&gt;モデルシステム

Amylopathy is a term that describes abnormal synthesis and accumulation of amyloid beta (A&beta;) in tissues with time. A&beta; is a hallmark of ...

A High Content Imaging Assay for Identification of Botulinum Neurotoxin Inhibitors

Synaptosomal-associated protein-25 (SNAP-25) is a component of the soluble NSF attachment protein receptor (SNARE) complex that is essential for synaptic neurotransmitter release. Botulinum neurotoxin serotype A (BoNT/A) is a zinc metalloprotease that blocks exocytosis of neurotransmitter by cleaving the SNAP-25 component of the SNARE complex. Currently there are no licensed medicines to treat BoNT/A poisoning after internalization of the toxin by motor neurons. The development of effective therapeutic measures to counter BoNT/A intoxication has been limited, due in part to the lack of robust high-throughput assays for screening small molecule libraries. Here we describe a high content imaging (HCI) assay with utility for identification of BoNT/A inhibitors. Initial optimization efforts focused on improving the reproducibility of inter-plate results across multiple, independent experiments. Automation of immunostaining, image acquisition, and image analysis were found to increase assay consistency and minimize variability while enabling the multiparameter evaluation of experimental compounds in a murine motor neuron system.

Botulinum neurotoxin is one of the most potent toxins among Category-A biothreat agents, yet a post-exposure therapeutic is not available. The high content imaging approach is a powerful methodology for identifying novel inhibitors as it enables multiparameter screening using biologically relevant motor neurons, the primary target of this toxin.

ボツリヌス神経毒素阻害剤の同定のためのハイコンテントイメージングアッセイ

Synaptosomal-associated protein-25 (SNAP-25) is a component of the soluble NSF attachment protein receptor (SNARE) complex that is essential for synaptic ...

Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits

The brain operates through the coordinated activation and the dynamic communication of neuronal assemblies. A major open question is how a vast repertoire of dynamical motifs, which underlie most diverse brain functions, can emerge out of a fixed topological and modular organization of brain circuits. Compared to in vivo studies of neuronal circuits which present intrinsic experimental difficulties, in vitro preparations offer a much larger possibility to manipulate and probe the structural, dynamical and chemical properties of experimental neuronal systems. This work describes an in vitro experimental methodology which allows growing of modular networks composed by spatially distinct, functionally interconnected neuronal assemblies. The protocol allows controlling the two-dimensional (2D) architecture of the neuronal network at different levels of topological complexity.
A desired network patterning can be achieved both on regular cover slips and substrate embedded micro electrode arrays. Micromachined structures are embossed on a silicon wafer and used to create biocompatible polymeric stencils, which incorporate the negative features of the desired network architecture. The stencils are placed on the culturing substrates during the surface coating procedure with a molecular layer for promoting cellular adhesion. After removal of the stencils, neurons are plated and they spontaneously redirected to the coated areas. By decreasing the inter-compartment distance, it is possible to obtain either isolated or interconnected neuronal circuits. To promote cell survival, cells are co-cultured with a supporting neuronal network which is located at the periphery of the culture dish. Electrophysiological and optical recordings of the activity of modular networks obtained respectively by using substrate embedded micro electrode arrays and calcium imaging are presented. While each module shows spontaneous global synchronizations, the occurrence of inter-module synchronization is regulated by the density of connection among the circuits.

This manuscript describes a protocol to grow in vitro modular networks consisting of spatially confined, functionally inter-connected neuronal circuits. A polymeric mask is used to pattern a protein layer to promote cellular adhesion over the culturing substrate. Plated neurons grow on coated areas establishing spontaneous connections and exhibiting electrophysiological activity.

デザイン、治療、セルラーめっき、機能的に相互接続された回路で構成モジュラー神経回路網の培養表面

The brain operates through the coordinated activation and the dynamic communication of neuronal assemblies. A major open question is how a vast repertoire ...

Analyzing Neural Activity and Connectivity Using Intracranial EEG Data with SPM Software

Measuring neural activity and connectivity associated with cognitive functions at high spatial and temporal resolutions is an important goal in cognitive neuroscience. Intracranial electroencephalography (EEG) can directly record electrical neural activity and has the unique potential to accomplish this goal. Traditionally, averaging analysis has been applied to analyze intracranial EEG data; however, several new techniques are available for depicting neural activity and intra- and inter-regional connectivity. Here, we introduce two analytical protocols we recently applied to analyze intracranial EEG data using the Statistical Parametric Mapping (SPM) software: time-frequency SPM analysis for neural activity and dynamic causal modeling of induced responses for intra- and inter-regional connectivity. We report our analysis of intracranial EEG data during the observation of faces as representative results. The results revealed that the inferior occipital gyrus (IOG) showed gamma-band activity at very early stages (110 ms) in response to faces, and both the IOG and amygdala showed rapid intra- and inter-regional connectivity using various types of oscillations. These analytical protocols have the potential to identify the neural mechanisms underlying cognitive functions with high spatial and temporal profiles.

We present two analytical protocols that can be used to analyze intracranial electroencephalography data using the Statistical Parametric Mapping (SPM) software: time-frequency statistical parametric mapping analysis for neural activity, and dynamic causal modeling of induced responses for intra- and inter-regional connectivity.

神経活動と Spm と頭蓋内脳波データを使用した接続の分析

Measuring neural activity and connectivity associated with cognitive functions at high spatial and temporal resolutions is an important goal in cognitive ...

Ex Vivo Oculomotor Slice Culture from Embryonic GFP-Expressing Mice for Time-Lapse Imaging of Oculomotor Nerve Outgrowth

Accurate eye movements are crucial for vision, but the development of the ocular motor system, especially the molecular pathways controlling axon guidance, has not been fully elucidated. This is partly due to technical limitations of traditional axon guidance assays. To identify additional axon guidance cues influencing the oculomotor nerve, an ex vivo slice assay to image the oculomotor nerve in real-time as it grows towards the eye was developed. E10.5 IslMN-GFP embryos are used to generate ex vivo slices by embedding them in agarose, slicing on a vibratome, then growing them in a microscope stage-top incubator with time-lapse photomicroscopy for 24-72 h. Control slices recapitulate the in vivo timing of outgrowth of axons from the nucleus to the orbit. Small molecule inhibitors or recombinant proteins can be added to the culture media to assess the role of different axon guidance pathways. This method has the advantages of maintaining more of the local microenvironment through which axons traverse, not axotomizing the growing axons, and assessing the axons at multiple points along their trajectory. It can also identify effects on specific subsets of axons. For example, inhibition of CXCR4 causes axons still within the midbrain to grow dorsally rather than ventrally, but axons that have already exited ventrally are not affected.

An ex vivo slice assay allows oculomotor nerve outgrowth to be imaged in real time. Slices are generated by embedding E10.5 IslMN:GFP embryos in agarose, slicing on a vibratome, and growing in a stage-top incubator. The role of axon guidance pathways is assessed by adding inhibitors to the culture media.

オキュロ運動神経増殖のタイムラプスイメージングのための胚性GFP発現マウスからのEx Vivo Oculomotorスライス培養

Accurate eye movements are crucial for vision, but the development of the ocular motor system, especially the molecular pathways controlling axon ...

Isolation and Culture of Oculomotor, Trochlear, and Spinal Motor Neurons from Prenatal Islmn:GFP Transgenic Mice

Oculomotor neurons (CN3s) and trochlear neurons (CN4s) exhibit remarkable resistance to degenerative motor neuron diseases such as amyotrophic lateral sclerosis (ALS) when compared to spinal motor neurons (SMNs). The ability to isolate and culture primary mouse CN3s, CN4s, and SMNs would provide an approach to study mechanisms underlying this selective vulnerability. To date, most protocols use heterogeneous cell cultures, which can confound the interpretation of experimental outcomes. To minimize the problems associated with mixed-cell populations, pure cultures are indispensable. Here, the first protocol describes in detail how to efficiently purify and cultivate CN3s/CN4s alongside SMNs counterparts from the same embryos using embryonic day 11.5 (E11.5) IslMN:GFP transgenic mouse embryos. The protocol provides details on the tissue dissection and dissociation, FACS-based cell isolation, and in vitro cultivation of cells from CN3/CN4 and SMN nuclei. This protocol adds a novel in vitro CN3/CN4 culture system to existing protocols and simultaneously provides a pure species- and age-matched SMN culture for comparison. Analyses focusing on the morphological, cellular, molecular, and electrophysiological characteristics of motor neurons are feasible in this culture system. This protocol will enable research into the mechanisms that define motor neuron development, selective vulnerability, and disease.

Isolation and Culture of Oculomotor, Trochlear, and Spinal Motor Neurons from Prenatal Islmn:GFP Transgenic Mice

This work presents a protocol to yield homogeneous cell cultures of primary oculomotor, trochlear, and spinal motor neurons. These cultures can be used for comparative analyses of the morphological, cellular, molecular, and electrophysiological characteristics of ocular and spinal motor neurons.

出生前イスルmnからのオキュロモータ、トロクレア、脊髄運動ニューロンの単離と培養 :GFPトランスジェニックマウス

Oculomotor neurons (CN3s) and trochlear neurons (CN4s) exhibit remarkable resistance to degenerative motor neuron diseases such as amyotrophic lateral ...