Questo lavoro è in parte supportato anche da finanziamenti National Institutes of Health (RO1 NS059043 e RO1 ES015988), Nazionale Sclerosi Multipla, la Roche Fondazione per la ricerca anemia, Feldstein Medical Foundation, e Shriners Hospitals for Children. Vorremmo ringraziare il Dr. Pleasure David per i suoi suggerimenti. Questo lavoro è stato in parte sostenuto dalle concessioni a WD dal National Institutes of Health (RO1 NS059043 e RO1 ES015988), Società Nazionale Sclerosi Multipla, e Shriners Hospitals for Children. Si dichiara non interessi contrastanti relativi a questo articolo.

Le procedure dettagliate di creare danni cerebrali neonatali nei topi P6: <ol><li> Postnatale giorno 6 (P6) sono topi anestetizzati con raffreddamento indiretto sul ghiaccio fino al punto di non responsività agli stimoli nocivi (anestesia profonda). </li><li> Dopo la pulizia della pelle con l&#39;alcol, una incisione mediana ventrale è fatto in parte anteriore del collo. </li><li> Sotto un microscopio da dissezione, la biforcazione della carotide comune destra si avvicina con delicatezza ritraendo i muscoli omoioideo e sternocleidomastoideo. </li><li> La fascia attorno alla guaina carotidea viene rimosso e il ramo prossimale interna isolata dal nervo vago e vicine gangli simpatici con un gancio. L&#39;arteria carotide interna viene cauterizzato con una punta di cauterizzare. </li><li> A seguito di cauterizzazione, l&#39;incisione cutanea viene suturata, e l&#39;animale è tenuto caldo fino a quando completamente sveglio e poi tornò alla diga. </li><li> Un&#39;ora dopo la legatura, l&#39;animale è posto in una camera sigillata infuso con azoto fino ad un livello del 6,0% si raggiunge O 2. L&#39;animale viene poi esposta a 35 minuti di ipossia. </li><li> Dopo l&#39;esposizione all&#39;ipossia, recupera il mouse su una piastra elettrica (33 ° C) per 30 minuti e poi viene restituito alla diga. Per la creazione di lesioni cerebrali con ipossia combinato / ischemia e di infezione / infiammazione, l&#39;animale viene quindi iniettato per via intraperitoneale con 0,015 ml di lipopolisaccaride (LPS, 1 mg / kg). </li><li> Quattro giorni post-hypoxia/ischemia, i topi vengono anestetizzati con raffreddamento indiretto sul ghiaccio fino al punto di non responsività agli stimoli nocivi, poi perfusi con soluzione salina e poi paraformaldeide 4%. Cervelli sono stati rimossi e crioprotetti. </li></ol><img src="/files/ftp_upload/1951/1951fig1.jpg" alt="Figura 1" /> Figura 1. Caratterizzazione di lesioni cerebrali nel ipossico / ischemico modello di topo con H &amp; E colorazione. Vi è necrosi focale (frecce) nella materia bianca cerebrale ipsilaterale (A) per la legatura della carotide, rispetto al controlaterale materia cerebrale bianca (B). Necrosi ipsilaterale e microinfarti focali sono osservati nell&#39;ippocampo (C) e il talamo (D) nel cervello con ipsilaterale lesioni della sostanza bianca cerebrale. Risultati Rappresentante e figure: Modelli murini di PVL con ipossia / ischemia e / o infezione / infiammazione si ottengono unilaterale legatura carotidea seguita da esposizione a ipossia e l&#39;iniezione di LPS. Stiamo definendo i modelli del mouse nuovi modelli adatti PVL. Esame neuropatologico di sezioni coronali colorate con ematossilina-and-eosina (H &amp; E) attraverso il proencefalo intero rivela necrosi focali nella materia bianca centrale dell&#39;emisfero cerebrale ipsilaterale alla legatura carotidea (Fig. 1A), con relativo risparmio di neuroni in la corteccia cerebrale sovrastante rispetto al controlaterale materia cerebrale bianca (Fig. 1B). Nella questione focale necrotico bianco, c&#39;è un marcato aumento della cellularità costituita da macrofagi e di astrociti reattivi, come è caratteristica della focale PVL umano. Nel controlaterale materia bianca centrale, i nuclei sono disposti in oligodendrogliali fascicolare-come fagotti, questa architettura è completamente distrutto dalla necrosi focale sul lato omolaterale. Da segnalare, inoltre osservare necrosi ipsilaterale nell&#39;ippocampo (Fig. 1C) e il talamo (microinfarti focale) (Fig. 1D) nel cervello con ipsilaterale lesioni della sostanza bianca cerebrale. Umano anche PVL non si verifica in completo isolamento dall&#39;ambiente accompagnano lesioni della sostanza grigia. Il segno distintivo della PVL è il relativo, anche se non completo, risparmiando della corteccia cerebrale sovrastante la materia bianca feriti - una caratteristica che noi crediamo è presente nei nostri modelli murini. <img src="/files/ftp_upload/1951/1951fig2.jpg" alt="Figura 2" /> Figura 2. Scala punteggio di 0-5 per la valutazione delle lesioni della sostanza bianca o MBP da O1 in modelli murini di leucomalacia periventricolare. I nostri modelli murini mostrano caratteristiche di PVL, sia a livello fenotipico e molecolare. Abbiamo usato il rilevamento immunocitochimica della proteina basica della mielina (MBP) o O1-antigene per valutare lesioni, hanno stabilito una scala standard da 0 a 5 a segnare il danno sostanza bianca (Fig. 2), e hanno caratterizzato la sostanza bianca patologia (Fig. . 3). Più recentemente, un test non-polarizzato stereological è stato implementato per sostituire il primo metodo semi-quantitativo. Inoltre, per testare gli effetti correlativo di lesioni della sostanza bianca sugli esiti funzionali, abbiamo svolto test comportamentali per la funzione motoria. Abbiamo dimostrato che i topi mostrano deficit motori colpendo P10-21 ipossia seguente / ischemia più LPS in P6. Abbiamo stabilito i criteri di punteggio con una scala di 0-4 per definire la disfunzione arto controlaterale nei topi feriti a P10-21. In genere, i topi normali può scorrazzano in pendenza di 30 ° (punteggio 4) e salire di 45 ° pendenza senza scivolare (punteggio di 3). Topi feriti hanno punteggi più bassi in correlazione con i gradi di lesioni della sostanza bianca. <img src="/files/ftp_upload/1951/1951fig3.jpg" alt="Figura 3" /> Figura 3. Caratterizzazione di lesioni della sostanza bianca o MBP da O1 colorazione e la struttura della mielina al microscopio elettronico (EM). MBP e la perdita di colorazione O1 spettacolo mielina nella materia bianca cerebrale ipsilaterale alla legatura, rispetto al controlaterale della sostanza bianca. Esame EM mostra assoni mielinizzati in materia bianca cerebrale controlaterale alla legatura e tessuto degenerato in ipsilaterale sostanza bianca. Noi prevediamo che la creazione dei modelli nuovo mouse di PVL ci permetterà di studiare meccanismi specifici di PVL-come lesioni al cervello immaturo. Ci si occuperà patogenesi utilizzando ceppi di topi transgenici per esaminare il ruolo delle singole molecole di interesse e valutare gli obiettivi di neuroprotezione.

<ol>
	<li>Volpe, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neurology of newborn. , 217-276 (2001).</li><li>Deng, W., Pleasure, J., Pleasure, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+periventricular+leukomalacia.">Progress in periventricular leukomalacia.</a> Arch. Neurol. 65, 1291-1295 (2008).</li><li>Follett, P. L., Deng, W., Dai, W., Talos, D. M., Massillon, L. J., Rosenberg, P. A., Volpe, J. J., Jensen, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutamate+receptor-mediated+oligodendrocyte+toxicity+in+periventricular+leukomalacia:+a+protective+role+for+topiramate.">Glutamate receptor-mediated oligodendrocyte toxicity in periventricular leukomalacia: a protective role for topiramate.</a> J. Neurosci. 24, 4412-4420 (2004).</li></ol>

Nessun conflitto di interessi dichiarati.

Infortunio per lo sviluppo del cervello porta a devastanti conseguenze neurologiche. Sorprendentemente, il modello di danno cerebrale perinatale è altamente dipendente dall&#39;età. In neonati a termine, colpisce prevalentemente corteccia cerebrale con perdita di neuroni caratteristici. Tuttavia, nei neonati prematuri, che attacca selettivamente la sostanza bianca cerebrale con danni importanti a oligodendrociti via di sviluppo, un disordine chiamato leucomalacia periventricolare (PVL). L&#39;entità del problema dei danni cerebrali nei neonati prematuri è straordinario. Parto prematuro e una migliore terapia intensiva neonatale hanno portato a quasi il 90% di sopravvivenza dei 13 milioni di neonati prematuri in tutto il mondo (circa 56.000 negli Stati Uniti) nasce ogni anno con un peso alla nascita inferiore a 1500 grammi, vale a dire, molto basso peso alla nascita i neonati. Circa il 10% dei sopravvissuti in seguito mostra la paralisi cerebrale e circa il 50% ha deficit cognitivi e comportamentali. PVL è il più importante paralisi cerebrale sottostante patologia cerebrale nei neonati prematuri. Nessuna terapia specifica per PVL esiste attualmente, in parte perché la patogenesi non è stato ben compreso 1,2. Sebbene l&#39;eziologia della PVL è multifattoriale, perinatale ipossia / ischemia con infezione materna intrauterina è pensato per essere una causa primaria di PVL. Un modello ipossico / ischemico della PVL nel ratto a P7 è stato stabilito precedentemente 3. Questo modello è unico in quanto assomiglia PVL nei neonati prematuri. La caratteristica di questo modello è selettiva sostanza bianca patologia, in contrasto con la maggior parte dei modelli ictus che sono caratterizzati da infarto materia grigia. Tuttavia, le limitazioni del modello ipossico-ischemica nel ratto non deve essere trascurato. In primo luogo, l&#39;eziologia della PVL è spesso multifattoriale. Oltre a ipossia / ischemia, materno-fetale dell&#39;infezione / infiammazione è fortemente associato con PVL. Così, insieme infiammazione ipossia / ischemia e di infezione / probabilmente creare modelli più PVL-like. L&#39;età roditore che meglio si correla con il periodo dello sviluppo umano di maggior rischio per le lesioni PVL (24-32 settimane di gestazione) è post-natale, perché la differenziazione oligodendrogliali è prevalentemente un evento postnatale nei roditori. Poiché è impossibile produrre un materno-fetale dell&#39;infezione / infiammazione in età post-natale, sarebbe ragionevole per simulare un&#39;infezione ad una rilevante fase di sviluppo post-natale per creare un modello di roditore di PVL. Abbiamo cercato di unire l&#39;iniezione LPS con il modello ipossico / ischemico nei topi a P6, di produrre lesioni PVL più clinicamente rilevante. In secondo luogo, l&#39;ottimale proof-of-concept approccio sperimentale per un ruolo causale di una molecola richiede l&#39;uso di topi transgenici, manipolazioni genetiche sono molto più difficili nel ratto che nel topo. Così, abbiamo cercato di convertire il nostro modello sperimentale di PVL dal topo al mouse. Modelli murini di PVL agevolerà notevolmente 1) lo studio della biologia della malattia e patogenesi utilizzando disponibili ceppi di topi transgenici, 2) conduzione di studi farmaco per scoprire agenti candidato potenziale terapeutico; 3) verifica dell&#39;utilità delle cellule staminali utilizzando ceppi di topi immunodeficienti.

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mouse models of periventricular leukomalacia

Abbiamo descritto un protocollo per stabilire modelli murini di leucomalacia periventricolare (PVL). PVL è la forma predominante di danno cerebrale nei neonati prematuri e l&#39;antecedente più comuni di paralisi cerebrale. PVL è caratterizzata da danno della sostanza bianca periventricolare con importanti lesioni oligodendrogliali. Ipossia / ischemia, con o senza infezione sistemica / infiammazione sono le cause primarie di PVL. Noi usiamo P6 topi per creare modelli di danno cerebrale neonatale con l&#39;induzione di ipossia / ischemia, con o senza infezione sistemica / infiammazione con legatura unilaterale della carotide seguiti da esposizione all&#39;ipossia, con o senza l&#39;iniezione di endotossina lipopolisaccaride (LPS). Immunoistochimica della proteina basica della mielina (MBP) o O1 e esame al microscopio elettronico mostrano perdita prominente mielina cerebrale nella materia bianca, con ulteriore danno per l&#39;ippocampo e talamo. Creazione di modelli murini di PVL faciliterà notevolmente lo studio della patogenesi della malattia utilizzando disponibili ceppi di topi transgenici, la conduzione di prove della droga in maniera velocità relativamente alta di identificare gli agenti terapeutici candidato, e la sperimentazione di trapianto di cellule staminali utilizzando ceppi di topi immunodeficienza.

Watch this Scientific Journal Video about Mouse Models of Periventricular Leukomalacia at JoVE.com

Mouse Models of Periventricular Leukomalacia

Abbiamo stabilito modelli murini di leucomalacia periventricolare (PVL), il danno cerebrale predominante in neonati prematuri caratterizzata da lesioni della sostanza bianca periventricolare. Ipossia / ischemia con / senza infezione sistemica sono le cause primarie del PVL. Unilaterale legatura carotidea e l&#39;esposizione ipossia con / senza iniezione lipopolisaccaride crea PVL-come le lesioni nei topi P6.

I modelli del mouse di leucomalacia periventricolare

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

mouse-models-of-periventricular-leukomalacia

Research

JoVE Journal

Neuroscience

29.7K Views.  University of California, Davis. Abbiamo stabilito modelli murini di leucomalacia periventricolare (PVL), il danno cerebrale predominante in neonati prematuri caratterizzata da lesioni della sostanza bianca periventricolare. Ipossia / ischemia con / senza infezione sistemica sono le cause primarie del PVL. Unilaterale legatura carotidea e l'esposizione ipossia con / senza iniezione lipopolisaccaride crea PVL-come le lesioni nei topi P6.

Video: Mouse Models of Periventricular Leukomalacia

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.

Un semplice composito Sistema di punteggio per la valutazione Fenotipo modelli murini di atassia cerebellare

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

Ricerca

Neuroscienze

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.

Multiple-topo Imaging neuroanatomici Risonanza Magnetica

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

Saggi olfattive per modelli murini di malattia di Neurodegenerative

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

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional units of the ventricles are ciliated epithelial cells termed ependymal cells, which line the ventricles and through ciliary action are capable of generating laminar flow of CSF at the ventricle surface. This monolayer of ependymal cells also provides barrier and filtration functions that promote exchange between brain interstitial fluids (ISF) and circulating CSF. Biochemical changes in the brain are thereby reflected in the composition of the CSF and destruction of the ependyma can disrupt the delicate balance of CSF and ISF exchange. In humans there is a strong correlation between lateral ventricle expansion and aging. Age-associated ventriculomegaly can occur even in the absence of dementia or obstruction of CSF flow. The exact cause and progression of ventriculomegaly is often unknown; however, enlarged ventricles can show regional and, often, extensive loss of ependymal cell coverage with ventricle surface astrogliosis and associated periventricular edema replacing the functional ependymal cell monolayer. Using MRI scans together with postmortem human brain tissue, we describe how to prepare, image and compile 3D renderings of lateral ventricle volumes, calculate lateral ventricle volumes, and characterize periventricular tissue through immunohistochemical analysis of en face lateral ventricle wall tissue preparations. Corresponding analyses of mouse brain tissue are also presented supporting the use of mouse models as a means to evaluate changes to the lateral ventricles and periventricular tissue found in human aging and disease. Together, these protocols allow investigations into the cause and effect of ventriculomegaly and highlight techniques to study ventricular system health and its important barrier and filtration functions within the brain.

Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain&#8217;s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.

Modellazione 3D del laterale ventricoli e istologica Caratterizzazione del Tissue periventricolare di esseri umani e il mouse

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional ...

Using Enzyme-based Biosensors to Measure Tonic and Phasic Glutamate in Alzheimer's Mouse Models

Neurotransmitter disruption is often a key component of diseases of the central nervous system (CNS), playing a role in the pathology underlying Alzheimer's disease, Parkinson's disease, depression, and anxiety. Traditionally, microdialysis has been the most common (lauded) technique to examine neurotransmitter changes that occur in these disorders. But because microdialysis has the ability to measure slow 1-20 minute changes across large areas of tissue, it has the disadvantage of invasiveness, potentially destroying intrinsic connections within the brain and a slow sampling capability. A relatively newer technique, the microelectrode array (MEA), has numerous advantages for measuring specific neurotransmitter changes within discrete brain regions as they occur, making for a spatially and temporally precise approach. In addition, using MEAs is minimally invasive, allowing for measurement of neurotransmitter alterations in vivo. In our laboratory, we have been specifically interested in changes in the neurotransmitter, glutamate, related to Alzheimer's disease pathology. As such, the method described here has been used to assess potential hippocampal disruptions in glutamate in a transgenic mouse model of Alzheimer's disease. Briefly, the method used involves coating a multi-site microelectrode with an enzyme very selective for the neurotransmitter of interest and using self-referencing sites to subtract out background noise and interferents. After plating and calibration, the MEA can be constructed with a micropipette and lowered into the brain region of interest using a stereotaxic device. Here, the method described involves anesthetizing rTg(TauP301L)4510 mice and using a stereotaxic device to precisely target sub-regions (DG, CA1, and CA3) of the hippocampus.

Using Enzyme-based Biosensors to Measure Tonic and Phasic Glutamate in Alzheimer&#039;s Mouse Models

Here, we describe the setup, software navigation, and data analysis for a spatially and temporally precise method of measuring tonic and phasic extracellular glutamate changes in vivo using enzyme-linked microelectrode arrays (MEA).

Utilizzando biosensori a base di enzimi misurare Tonic e Phasic glutammato in modelli murini di Alzheimer

Neurotransmitter disruption is often a key component of diseases of the central nervous system (CNS), playing a role in the pathology underlying ...

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this challenging. Because the ventricles are negative spaces located deep within the brain, the only way to understand their anatomy is by appreciating their boundaries formed by related structures. Looking at a 2D representation of these spaces, in any of the cardinal planes, will not enable visualisation of all of the structures that form the boundaries of the ventricles. Thus, using 2D sections alone requires students to compute their own mental image of the 3D ventricular spaces. The aim of this study was to develop a reproducible method for dissecting the human brain to create an educational resource to enhance student understanding of the intricate relationships between the ventricles and periventricular structures. To achieve this, we created a video resource that features a step-by-step guide using a fiber dissection method to reveal the lateral and third ventricles together with the closely related limbic system and basal ganglia structures. One of the advantages of this method is that it enables delineation of the white matter tracts that are difficult to distinguish using other dissection techniques. This video is accompanied by a written protocol that provides a systematic description of the process to aid in the reproduction of the brain dissection. This package offers a valuable anatomy teaching resource for educators and students alike. By following these instructions educators can create teaching resources and students can be guided to produce their own brain dissection as a hands-on practical activity. We recommend that this video guide be incorporated into neuroanatomy teaching to enhance student understanding of the morphology and clinical relevance of the ventricles.

This paper demonstrates the effective use of a fiber dissection method to reveal the superficial white matter tracts and periventricular structures of the human brain, in three-dimensional space, to aid student comprehension of ventricular morphology.

Esplorazione dello spazio profondo - scoprendo l'anatomia delle strutture Periventricular per rivelare i ventricoli laterali del cervello umano

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

Establishing Mouse Models for Zika Virus-induced Neurological Disorders Using Intracerebral Injection Strategies: Embryonic, Neonatal, and Adult

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly and additional brain abnormalities. However, the mechanism underlying the pathogenesis of ZIKV in the developing brain remains unclear. Intracerebral surgical methods are frequently used in neuroscience research to address questions about both normal and abnormal brain development and brain function. This protocol utilizes classical surgical techniques and describes methods that allow one to model ZIKV-associated human neurological disease in the mouse nervous system. While direct brain inoculation does not model the normal mode of virus transmission, the method allows investigators to ask targeted questions concerning the consequence after ZIKV infection of the developing brain. This protocol describes embryonic, neonatal, and adult stages of intraventricular inoculation of ZIKV. Once mastered, this method can become a straightforward and reproducible technique that only takes a few hours to perform.

Here we describe a method for establishing a model of Zika virus-induced microcephaly in mouse. This protocol includes methods for embryonic, neonatal, and adult-stage intracerebral inoculation of the Zika virus.

Creazione di modelli murini per disturbi neurologici indotta da Virus di Zika utilizzando strategie di iniezione intracerebrale: embrionale, neonatale e adulto

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly ...

Adult Mouse DRG Explant and Dissociated Cell Models to Investigate Neuroplasticity and Responses to Environmental Insults Including Viral Infection

This protocol describes an ex vivo model of mouse-derived dorsal root ganglia (DRG) explant and in vitro DRG-derived co-culture of dissociated sensory neurons and glial satellite cells. These are useful and versatile models to investigate a variety of biological responses associated with physiological and pathological conditions of the peripheral nervous system (PNS) ranging from neuron-glial interaction, neuroplasticity, neuroinflammation, and viral infection. The usage of DRG explant is scientifically advantageous compared to simplistic single cells models for multiple reasons. For instance, as an organotypic culture, the DRG explant allows ex vivo transfer of an entire neuronal network including the extracellular microenvironment that play a significant role in all the neuronal and glial functions. Further, DRG explants can also be maintained ex vivo for several days and the culture conditions can be perturbed as desired. In addition, the harvested DRG can be further dissociated into an in vitro co-culture of primary sensory neurons and satellite glial cells to investigate neuronal-glial interaction, neuritogenesis, axonal cone interaction with the extracellular microenvironment, and more general, any aspect associated with the neuronal metabolism. Therefore, the DRG-explant system offers a great deal of flexibility to study a wide array of events related to biological, physiological, and pathological conditions in a cost-effective manner.

In this report, the advantages of organotypic cultures and dissociated primary cultures of mouse-derived dorsal root ganglia are highlighted to investigate a wide range of mechanisms associated with neuron-glial interaction, neuroplasticity, neuroinflammation, and response to viral infection.

Topo adulto DRG Explant e dissociato modelli cellulari per studiare neuroplasticità e risposte agli insulti ambientali, compreso l'infezione virale

This protocol describes an ex vivo model of mouse-derived dorsal root ganglia (DRG) explant and in vitro DRG-derived co-culture of dissociated sensory ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings provide a sensitive approach to measure nerve conduction in humans and rodent models. To broaden the technical possibilities for electromyography in mice, the measurement of compound muscle action potentials (CMAPs) from the brachial plexus nerve in the forelimb using needle electrodes is described here. CMAP recordings after stimulating the sciatic nerve in hindlimbs have been previously described. The newly introduced method here allows for the evaluation of the nerve conductivity at an additional site, and thus provides a more profound overview of the neuromuscular functionality. The technique provides information on both the relative number of functional axons and the myelination level. Thereby, this method can be applied to assess both axonal diseases as well as demyelinating conditions. This minimally invasive method does not require extraction of the nerve and therefore it is suitable for repeated measurements for longitudinal follow-up in the same animal. Similar recordings are performed in clinical setups to emphasize the translational relevance of the method.

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

In Vivo Elettrofisiologici misurazione del potenziale di azione muscolare composto da arti anteriori in modelli murini di degenerazione del motoneurone

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...

A Behavioral Screen for Heat-Induced Seizures in Mouse Models of Epilepsy

Transgenic mouse models have proved to be powerful tools in studying various aspects of human neurological disorders, including epilepsy. The SCN1A-associated genetic epilepsies comprise a wide spectrum of seizure disorders with incomplete penetrance and clinical variability. SCN1A mutations can result in a large variety of seizure phenotype ranging from simple, self-limited fever-associated febrile seizures (FS), moderate-level genetic epilepsy with febrile seizures plus (GEFS+) to more severe Dravet Syndrome (DS). Although FS are commonly seen in children below 6-7 years of age who do not have genetic epilepsy, FS in GEFS+ patients continue to occur into adulthood. Traditionally, experimental FS have been induced in mice by exposing the animal to a stream of dry air or heating lamps, and the rate of change in body temperature is often not well controlled. Here, we describe a custom-built heating chamber, with a plexiglass front, that is fitted with a digital temperature controller and a heater-equipped electric fan, which can send heated forced air into the test arena in a temperature-controlled manner. The body temperature of a mouse placed in the chamber, monitored through a rectal probe, can be increased to 40-42 &#176;C in a reproducible manner by increasing the temperature inside the chamber. Continual visual monitoring of the animals during the heating period demonstrates induction of heat-induced seizures in mice carrying an FS mutation at a body temperature that does not elicit behavioral seizures in wild-type litter mates. Animals can be easily removed from the chamber and placed on a cooling pad to rapidly return body temperature to normal. This method provides for a simple, rapid, and reproducible screening protocol for the occurrence of heat-induced seizures in epilepsy mouse models.

The goal of the method is to screen for hyperthermia or heat-induced seizures in mouse models. The protocol describes the use of a custom-built chamber with continuous monitoring of the body temperature to determine whether elevated body temperature leads to seizures.