この作品は、GAGとEY014061＆EY014997 ARLへ、だけでなく、UL1DE019583 - 02コンソーシアム賞にNIHの助成金NS052535によってサポートされていました。

バイアスを防ぐために、評価を行う実験者が動物の遺伝子型の知識を持つべきではありません。個々の対策が最も重篤な症状を表す、関連する表現型と3の非存在を表す0で、0〜3のスケールで採点されます。各テストは、スコアが再現可能であることを保証するために複数回実行されます。肥満は記述されているすべての措置の解釈が複雑になります。研究者は結果で肥満の役割の可能性を評価するための表現型のスコアリング後のマウスを比較検討することもできます。 レッジのテストレッジのテストでは、小脳性運動失調および他の多くの神経変性疾患で損なわれている調整の直接的な指標である。この措置は、小脳性運動失調の人間の兆候に最も直接的に比較です。 <ol><li>ケージからマウスを持ち上げて、ケージの棚の上に置きます。マウスは、通常は棚に沿って歩くとケージに戻って下降しようとします。 </li><li>それはケージの棚に沿って歩くと、そのケージに自身を低下させるとして、マウスを観察。野生型マウスでは、その足を使って、通常はそのバランスを失うことなく、棚に沿って歩いていく、そして優雅にケージに自身を戻って低下します。これは、0のスコアが割り当てられます。マウスは棚に沿って歩いている間、その足場を失うが、それ以外は調整が表示された場合、それは1のスコアを受け取ります。ケージに下降したときに効果的に頭ではなく、その足で、その後ろ足、または土地を使用しない場合、それは2のスコアを受け取ります。それが棚から落ちた、またはそれに近い、徒歩またはそれ自体を下げるしようとしたとき、または揺れや励ましにもかかわらず、全く移動を拒否した場合、それは3のスコアを受け取ります。一部のマウスは、それらを棚に沿って歩いたり、ケージの中に下降することを奨励するために穏やかなナッジが必要になります。 </li><li>レッジのテストの得点を記録。 </li></ol>抱茎の後肢抱茎の後肢は、特定の小脳性運動失調[1]を含む神経変性モデルマウス、数の疾患の進行のマーカーである。 <ol><li>そのベースの近くに尾をつかんで、すべての周囲のオブジェクトの透明なマウスを持ち上げる。 </li><li> 10秒間後肢の位置を守ってください。後肢が一貫して腹部から離れて、外側にスプレイされている場合、それは0のスコアが割り当てられます。 one後肢が中断時間の50％以上を腹部に向かって後退している場合、それは1のスコアを受け取ります。両方の後肢の一部が一時停止時間の50％以上を腹部に向かって収納されている場合、それは2のスコアを受け取ります。その後肢は完全に撤回し、中断時間の50％以上を腹部に触れている場合、それは3のスコアを受け取ります。 </li><li>そのケージに戻すマウスを置き、その後肢抱茎のスコアを記録。 </li></ol>足取り歩行は、調整と筋肉の機能の指標です。 <ol><li>そのケージからマウスを削除し、その頭が離れて研究者から上向きにして平らな面に置きます。 </li><li>それは歩くと後ろからマウスを守ってください。マウスがその体重がその腹部が地面に触れていないと、すべての手足でサポートされている、と両方後肢が均等に参加して、正常に動くなら、それは0のスコアを受け取ります。それは振戦を示したり、歩きながら足を引きずるように見える場合、それは1のスコアを受け取ります。それが表示される場合は重度の振戦、重度の跛行は、骨盤を下げたり、足が移動（&quot;鴨足&quot;）の間に身体から離れて指し、それは2のスコアを受け取ります。マウスが困難前進があり、地面に沿って腹部をドラッグしたら、それは3のスコアを受け取ります。 </li><li>そのケージに戻すマウスを配置し、歩行のスコアを記録。 </li></ol>脊柱後弯症脊柱後弯症は、マウスモデルの神経変性疾患の共通の症状[2]である背骨の特徴背曲です。それは、神経変性への二次的脊椎筋肉の筋緊張の消失によって引き起こされている。 <ol><li>そのケージからマウスを取り外し、平らな面に置きます。 </li><li>それが歩くとき、それを守ってください。マウスはそれが歩くとき、簡単にその背骨をまっすぐにすることができる、そして永続的な脊柱後がない場合は、それは0のスコアを受け取ります。マウスは、軽度の脊柱後弯症を示すが、その背骨をまっすぐにすることができるなら、それは1のスコアを受け取ります。それは完全に背骨をまっすぐにすることができないと永続的なしかし、軽度の脊柱後弯症を保持している場合、それは2のスコアを受け取ります。それが歩くときや、それが座っている間、マウスは顕著な脊柱後弯症を維持する場合、それは3のスコアが割り当てられます。 </li><li>そのケージに戻すマウスを置き、その脊柱後のスコアを記録。 </li></ol>代表的な結果動物の十分な数で、この手順では、系統間と時間をかけて同じ系統内で表現型の違いを検出することができる。データ分析のために、とることによって各数値データのスコアを計算する個々の三測定の平均値。措置は、別に分析したり、個々の複合表現型のスコアにまとめることができます。 データをグラフ化し、重要性を判断するために適切な統計的手法を適用する。図1は、マウスのトランスジェニックSCA7モデルの複合表現型の評価の結果を示しています。このfloxed - SCA7 - 92Qトランスジェニックモデルでは、92のCAGリピートを持つ人間ataxin - 7遺伝子は、つのloxP部位に隣接されており、細菌人工染色体から発現。複合表現型のスコアは12の可能な最高得点のための抱茎の、レッジの歩行、歩行や脊柱後の評価が含まれています。 floxed - SCA7 - 92QトランスジェニックマウスにおけるプログレッシブSCA7表現型はヒトの疾患の進行性の性質と一致しているの増加の複合表現型のスコアによって示されている。 <img src="/files/ftp_upload/1787/1787fig1.jpg" alt="図1" /> 図1 Floxed - SCA7 - 92Qマウスは20週（：; *** P &lt;0.001ボンフェローニ事後 2ウェイANOVA）から始まる非トランスジェニック同腹子と著しく異なるプログレッシブSCA7表現型を示す。マウスは、歩行、および脊柱後を抱茎の、岩棚のテストのための0から3のスケールごとに評価した。各年齢における各遺伝子型の平均コンポジットス​​コアを算出した。バーはSEMを示す。

<ol>
	<li>Chou, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polyglutamine-expanded+ataxin-3+causes+cerebellar+dysfunction+of+SCA3+transgenic+mice+by+inducing+transcriptional+dysregulation.">Polyglutamine-expanded ataxin-3 causes cerebellar dysfunction of SCA3 transgenic mice by inducing transcriptional dysregulation.</a> Neurobiol Dis. 31 (1), 89-101 (2008).</li><li>Thomas, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+endogenous+androgen+receptor+protein+accelerates+motor+neuron+degeneration+and+accentuates+androgen+insensitivity+in+a+mouse+model+of+X-linked+spinal+and+bulbar+muscular+atrophy.">Loss of endogenous androgen receptor protein accelerates motor neuron degeneration and accentuates androgen insensitivity in a mouse model of X-linked spinal and bulbar muscular atrophy.</a> Hum Mol Genet. 15 (14), 2225-2238 (2006).</li><li>Ditzler, S., Stoeck, J., LeBlanc, M., Kooperberg, C., Hansen, S., Coppin, L., Olson, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Rapid+Neurobehavioral+Assessment+Reveals+that+FK506+Delays+Symptom+Onset+in+R6/2+Huntington's+Disease+Mice.">A Rapid Neurobehavioral Assessment Reveals that FK506 Delays Symptom Onset in R6/2 Huntington's Disease Mice.</a> Preclinica Research Articles. 1 (3), 115-126 (2003).</li></ol>

これらの研究は、ワシントン動物実験の大学および使用委員会によって承認された。

このプロトコルは、小脳性運動失調のマウスモデルにおける疾患の重症度の高感度かつ容易に行う評価できるように設計されています。スコアリングシステムの個々のコンポーネントは、神経変性の異なるマウスモデルで多かれ少なかれ効果的になります。 このスコアリングシステムの要素を効果的に小脳失調症、ハンチントン病と球脊髄性筋萎縮症[1-3]を含むヒトの神...

a simple composite phenotype scoring system for evaluating mouse models of cerebellar ataxia

私たちは、小脳性運動失調症のマウスモデルにおける疾患の重症度の迅速かつ高感度定量のためのプロトコルについて説明します。それは脊髄小脳変性症、ハンチントン病と球脊髄性筋萎縮症を含むいくつかの疾患モデルで以前に発行された表現型の評価、から派生しています。対策は、後肢抱茎の、レッジの試験、歩行や脊柱後などがあります。各指標は、4つのすべての措置に対して0〜12の合計で、0〜3のスケールで記録されます。また、神経変性疾患の表現型の時間的進行を定量化しながら結果は事実上、影響を受けると非罹患個体との間で区別する。対策は、個々に分析さ以上の統計的検出力のための複合表現型のスコアに組み合わせることができる。 four説明した対策の理想的な組み合わせは、問題の疾患に依存します。我々は、脊髄小脳失調タイプ7（SCA7）のトランスジェニックマウスモデルでの疾患の重症度を評価するために使用されるプロトコルの例を示す。 アルバートR.ラスパーダとグウェンA.庭は均等にこの原稿に貢献した。

Watch this Scientific Journal Video about A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia at JoVE.com

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

私たちは、小脳性運動失調症のマウスモデルにおける疾患の重症度の迅速かつ高感度定量のためのプロトコルについて説明します。対策は、後肢抱茎の、レッジの試験、歩行や脊柱後などがあります。このプロトコルは、効果的に影響を受けたと非罹患個体を区別し、時間の経過に影響を受ける個人の進行を検出する。

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 ...

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Research

JoVE Journal

Neuroscience

35.4K ビュー.  University of Washington. 私たちは、小脳性運動失調症のマウスモデルにおける疾患の重症度の迅速かつ高感度定量のためのプロトコルについて説明します。対策は、後肢抱茎の、レッジの試験、歩行や脊柱後などがあります。このプロトコルは、効果的に影響を受けたと非罹患個体を区別し、時間の経過に影響を受ける個人の進行を検出する。

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

Mouse Models of Periventricular Leukomalacia

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature infants and the most common antecedent of cerebral palsy. PVL is characterized by periventricular white matter damage with prominent oligodendroglial injury. Hypoxia/ischemia with or without systemic infection/inflammation are the primary causes of PVL. We use P6 mice to create models of neonatal brain injury by the induction of hypoxia/ischemia with or without systemic infection/inflammation with unilateral carotid ligation followed by exposure to hypoxia with or without injection of the endotoxin lipopolysaccharide (LPS). Immunohistochemistry of myelin basic protein (MBP) or O1 and electron microscopic examination show prominent myelin loss in cerebral white matter with additional damage to the hippocampus and thalamus. Establishment of mouse models of PVL will greatly facilitate the study of disease pathogenesis using available transgenic mouse strains, conduction of drug trials in a relatively high throughput manner to identify candidate therapeutic agents, and testing of stem cell transplantation using immunodeficiency mouse strains.

We established mouse models of periventricular leukomalacia (PVL), the predominant brain injury in premature infants characterized by periventricular white matter lesions. Hypoxia/ischemia with/without systemic infection are the primary causes of PVL. Unilateral carotid ligation and hypoxia exposure with/without lipopolysaccharide injection creates PVL-like lesions in P6 mice.

脳室周囲白質軟化症のマウスモデル

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature ...

神経科学

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

複数のマウスの神経解剖学的磁気共鳴イメージング

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

Organotypic Cerebellar Cultures: Apoptotic Challenges and Detection

Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 1,2 and have constituted a major breakthrough in the field of neuroscience. Since then, the technique was developed further and currently there are many different ways to prepare organotypic cultures. The method presented here was adapted from the one described by Stoppini et al. for the preparation of the slices and from Gogolla et al. for the staining procedure 3,4.
A unique feature of this technique is that it allows you to study different parts of the brain such as hippocampus or cerebellum in their original structure, providing a big advantage over dissociated cultures in which all the cellular organization and neuronal networks are disrupted. In the case of the cerebellum it is even more advantageous because it allows the study of Purkinje cells, extremely difficult to obtain as dissociated primary culture. This method can be used to study certain developmental features of the cerebellum in vitro, as well as for electrophysiological and pharmacological experiments in both wild type and mutant mice.
The method described here was designed to study the effect of apoptotic stimuli such as Fas ligand in the developing cerebellum, using TUNEL staining to measure apoptotic cell death. If TUNEL staining is combined with cell type specific markers, such as Calbindin for Purkinje cells, it is possible to evaluate cell death in a cell population specific manner. The Calbindin staining also serves the purpose of evaluating the quality of the cerebellar cultures.

This method describes the generation of organotypic cerebellar cultures and the effect of certain apoptotic stimuli on the viability of different cerebellar cell types.

器官小脳文化：アポトーシス課題と検出

Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 1,2 and have constituted a major breakthrough in the field of neuroscience. ...

Quantitative Analysis of Synaptic Vesicle Pool Replenishment in Cultured Cerebellar Granule Neurons using FM Dyes

After neurotransmitter release in central nerve terminals, SVs are rapidly retrieved by endocytosis. Retrieved SVs are then refilled with neurotransmitter and rejoin the recycling pool, defined as SVs that are available for exocytosis1,2. The recycling pool can generally be subdivided into two distinct pools - the readily releasable pool (RRP) and the reserve pool (RP). As their names imply, the RRP consists of SVs that are immediately available for fusion while RP SVs are released only during intense stimulation1,2. It is important to have a reliable assay that reports the differential replenishment of these SV pools in order to understand 1) how SVs traffic after different modes of endocytosis (such as clathrin-dependent endocytosis and activity-dependent bulk endocytosis) and 2) the mechanisms controlling the mobilisation of both the RRP and RP in response to different stimuli.
FM dyes are routinely employed to quantitatively report SV turnover in central nerve terminals3-8. They have a hydrophobic hydrocarbon tail that allows reversible partitioning in the lipid bilayer, and a hydrophilic head group that blocks passage across membranes. The dyes have little fluorescence in aqueous solution, but their quantum yield increases dramatically when partitioned in membrane9. Thus FM dyes are ideal fluorescent probes for tracking actively recycling SVs. The standard protocol for use of FM dye is as follows. First they are applied to neurons and are taken up during endocytosis (Figure 1). After non-internalised dye is washed away from the plasma membrane, recycled SVs redistribute within the recycling pool. These SVs are then depleted using unloading stimuli (Figure 1). Since FM dye labelling of SVs is quantal10, the resulting fluorescence drop is proportional to the amount of vesicles released. Thus, the recycling and fusion of SVs generated from the previous round of endocytosis can be reliably quantified.
Here, we present a protocol that has been modified to obtain two additional elements of information. Firstly, sequential unloading stimuli are used to differentially unload the RRP and the RP, to allow quantification of the replenishment of specific SV pools. Secondly, each nerve terminal undergoes the protocol twice. Thus, the response of the same nerve terminal at S1 can be compared against the presence of a test substance at phase S2 (Figure 2), providing an internal control. This is important, since the extent of SV recycling across different nerve terminals is highly variable11.
Any adherent primary neuronal cultures may be used for this protocol, however the plating density, solutions and stimulation conditions are optimised for cerebellar granule neurons (CGNs)12,13.

A live fluorescence imaging technique to quantify the replenishment and mobilisation of specific synaptic vesicle (SV) pools in central nerve terminals is described. Two rounds of SV recycling are monitored in the same nerve terminals providing an internal control.

FM染料を用いて培養小脳顆粒神経細胞のシナプス小胞プールの補充の定量分析

After neurotransmitter release in central nerve terminals, SVs are rapidly retrieved by endocytosis.  Retrieved SVs are then refilled with ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

ラットのショ糖の渇望のインキュベーションを評価するための一般的な方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Olfactory Assays for Mouse Models of Neurodegenerative Disease

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process and may be a useful behavioral marker for early detection. Earlier detection in neurodegenerative disease is a major goal in the field because this is when neuroprotective therapies have the best potential to be effective. Therefore, in preclinical studies testing novel neuroprotective strategies in rodent models of neurodegenerative disease, olfactory assessment could be highly useful in determining therapeutic potential of compounds and translation to the clinic. In the present study we describe a battery of olfactory assays that are useful in measuring olfactory function in mice. The tests presented in this study were chosen because they measure olfaction abilities in mice related to food odors, social odors, and non-social odors. These tests have proven useful in characterizing novel genetic mouse models of Parkinson&#8217;s disease as well as in testing potential disease-modifying therapies.

Impairment in olfactory function is a common feature in many neurodegenerative disorders including Parkinson, Alzheimer, and Huntington diseases. In the present article, we describe a set of tests for assessing olfaction discrimination and detection in mice that can be used to measure olfactory abilities in mouse models of neurodegenerative diseases.

神経変性疾患のマウスモデルのための嗅覚アッセイ

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process ...

Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons

Cerebellar granule neurons (CGNs) are a commonly used neuronal model, forming an abundant homogeneous population in the cerebellum. In light of their post-natal development, abundance, and accessibility, CGNs are an ideal model to study neuronal processes, including neuronal development, neuronal migration, and physiological neuronal activity stimulation. In addition, CGN cultures provide an excellent model for studying different modes of cell death including excitotoxicity and apoptosis. Within a week in culture, CGNs express N-methyl-D-aspartate (NMDA) receptors, a specific ionotropic glutamate receptor with many critical functions in neuronal health and disease. The addition of low concentrations of NMDA in conjunction with membrane depolarization to rodent primary CGN cultures has been used to model physiological neuronal activity stimulation while the addition of high concentrations of NMDA can be employed to model excitotoxic neuronal injury. Here, a method of isolation and culturing of CGNs from 6 day old pups as well as genetic manipulation of CGNs by adenoviruses and lentiviruses are described. We also present optimized protocols on how to stimulate NMDA-induced excitotoxicity, low-potassium-induced apoptosis, oxidative stress and DNA damage following transduction of these neurons.

This protocol describes a simple method for isolating and culturing primary mouse cerebral granule neurons (CGNs) from 6-7 day old pups, efficient transduction of CGNs for loss and gain of function studies, and modelling NMDA-induced neuronal excitotoxicity, low-potassium-induced cell death, DNA-damage, and oxidative stress using the same culture model.

神経細胞死とマウス プライマリ小脳顆粒ニューロンの変性をモデリング

Cerebellar granule neurons (CGNs) are a commonly used neuronal model, forming an abundant homogeneous population in the cerebellum. In light of their ...

Transplantation of Chemogenetically Engineered Cortical Interneuron Progenitors into Early Postnatal Mouse Brains

Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is a complicated task, which is particularly difficult in regards to the development of &#947;-aminobutyric acid (GABA)ergic cortical interneurons (CIs). CIs are the main inhibitory neurons in the cerebral cortex, and they play key roles in neuronal networks, by regulating both the activity of individual pyramidal neurons, as well as the oscillatory behavior of neuronal ensembles. They are generated in transient embryonic structures (medial and caudal ganglionic eminences -&#160;MGE and CGE) that are very difficult to efficiently target using in utero electroporation approaches. Interneuron progenitors migrate long distances during normal embryonic development, before they integrate in the cortical circuit. This remarkable ability to disperse and integrate into a developing network can be hijacked by transplanting embryonic interneuron precursors into early post-natal host cortices. Here, we present a protocol that allows genetic modification of embryonic interneuron progenitors using focal ex vivo electroporation. These engineered interneuron precursors are then transplanted into early post-natal host cortices, where they will mature into easily identifiable CIs. This protocol allows the use of multiple genetically encoded tools, or the ability to regulate the expression of specific genes in interneuron progenitors, in order to investigate the impact of either genetic or environmental variables on the maturation and integration of CIs.

Here we present a protocol, designed to use chemogenetic tools to manipulate the activity of cortical interneuron progenitors transplanted into the cortex of early postnatal mice.

化学遺伝子解析で作られた皮質間ニューロン前駆体を早期産後マウス脳に移植

Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is ...

Cerebellar Regional Dissection for Molecular Analysis

Cerebellum plays an important role in several key functions including control of movement, balance, cognition, reward, and affect. Imaging studies indicate that distinct cerebellar regions contribute to these different functions. Molecular studies examining regional cerebellar differences are lagging as they are mostly done on whole cerebellar extracts thereby masking any distinctions across specific cerebellar regions. Here we describe a technique to reproducibly and quickly dissect four different cerebellar regions: the deep cerebellar nuclei (DCN), anterior and posterior vermal cerebellar cortex, and the cerebellar cortex of the hemispheres. Dissecting out these distinct regions allows for the exploration of molecular mechanisms that may underlie their unique contributions to balance, movement, affect and cognition. This technique may also be used to explore differences in pathological susceptibility of these specific regions across various mouse disease models.

Different cerebellar regions have been implicated to play a role in distinct behavioral outputs, yet the underlying molecular mechanisms remain unknown. This work describes a method to reproducibly and quickly dissect cerebellar cortex of the hemispheres, anterior and posterior regions of the vermis, and the deep cerebellar nuclei in order to probe for molecular differences by isolating RNA and testing for differences in gene expression.&#160;

分子解析のための小脳局所解剖

Cerebellum plays an important role in several key functions including control of movement, balance, cognition, reward, and affect. Imaging studies ...

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor impairment that worsens with time. While considerable advances have been made in determining genetic drivers of ALS for a subset of patients, the majority of cases have an unknown etiology. Further, the mechanisms underlying motor neuron dysfunction and degeneration are not well understood; therefore, there is an ongoing need to develop and characterize representative models to study these processes. Caenorhabditis elegans can adapt their movement to the physical constraints of their surroundings, with two primary movement paradigms studied in a laboratory environment- crawling on a solid surface and swimming in liquid. These represent a complex interplay between sensation, motor neurons, and muscles. C. elegans models of ALS can exhibit impairment in one or both of these movement paradigms. This protocol describes two sensitive assays for evaluating motility in C. elegans: an optimized radial locomotion assay measuring crawling on a solid surface and an automated method for tracking and analyzing swimming in liquid (thrashing). In addition to the characterization of baseline motor impairment of ALS models, these assays can detect suppression or enhancement of the phenotypes from genetic or small molecule interventions. Thus, these methods have utility for studying ALS&#160;models and any C. elegans strain that exhibits altered motility.

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

This protocol describes two sensitive assays for discriminating among mild, moderate, and severe motor impairment in C. elegans models of amyotrophic lateral sclerosis, with general utility for C. elegans strains, with altered motility.

筋萎縮性側索硬化症の C.エレガンス モデルにおける運動障害の評価

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor ...

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

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この動画の章

脳室周囲白質軟化症のマウスモデル

複数のマウスの神経解剖学的磁気共鳴イメージング

器官小脳文化：アポトーシス課題と検出

FM染料を用いて培養小脳顆粒神経細胞のシナプス小胞プールの補充の定量分析

ラットのショ糖の渇望のインキュベーションを評価するための一般的な方法

神経変性疾患のマウスモデルのための嗅覚アッセイ

神経細胞死とマウスプライマリ小脳顆粒ニューロンの変性をモデリング

化学遺伝子解析で作られた皮質間ニューロン前駆体を早期産後マウス脳に移植

分子解析のための小脳局所解剖

筋萎縮性側索硬化症の C.エレガンスモデルにおける運動障害の評価

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

要約

さらに動画を探す

この動画の章

脳室周囲白質軟化症のマウスモデル

複数のマウスの神経解剖学的磁気共鳴イメージング

器官小脳文化：アポトーシス課題と検出

FM染料を用いて培養小脳顆粒神経細胞のシナプス小胞プールの補充の定量分析

ラットのショ糖の渇望のインキュベーションを評価するための一般的な方法

神経変性疾患のマウスモデルのための嗅覚アッセイ

神経細胞死とマウス プライマリ小脳顆粒ニューロンの変性をモデリング

化学遺伝子解析で作られた皮質間ニューロン前駆体を早期産後マウス脳に移植

分子解析のための小脳局所解剖

筋萎縮性側索硬化症の C.エレガンス モデルにおける運動障害の評価

神経細胞死とマウスプライマリ小脳顆粒ニューロンの変性をモデリング

筋萎縮性側索硬化症の C.エレガンスモデルにおける運動障害の評価