Name: mouse models of periventricular leukomalacia
Uploaded: 2010-05-18T17:00:00
Duration: 6 min 24 s
Description: Watch this Scientific Journal Video about Mouse Models of Periventricular Leukomalacia at JoVE.com

Detta arbete är delvis finansierats med bidrag från National Institutes of Health (RO1 NS059043 och RO1 ES015988), National Multiple Sclerosis Society, Roche Stiftelsen för anemi Forskning, Feldstein Medical Foundation och Shriners sjukhus för barn. Vi vill tacka Dr David Njutning för hans förslag. Detta arbete delvis finansierats med bidrag till WD från National Institutes of Health (RO1 NS059043 och RO1 ES015988), National Multiple Sclerosis Society, och Shriners sjukhus för barn. Vi förklarar inga konkurrerande intressen med anknytning till denna artikel.

Detaljerade förfaranden för att skapa neonatal hjärnskada i P6 möss: <ol><li> Postnatal dag 6 (P6) möss bedövas med indirekt kylning på is till den grad att inga reaktioner på skadliga stimulering (djup narkos). </li><li> Efter rengöring av huden med alkohol, är ett mittlinjen ventrala snitt i främre halsen. </li><li> Enligt en dissekera mikroskop, är bifurkation av rätten gemensamma halspulsådern närmade sig genom att försiktigt dra tillbaka omohyoid och sternocleidomastoideus musklerna. </li><li> Fascian runt halspulsådern slida tas bort och den proximala interna gren isolerade från den närliggande vagusnerven och sympatiska ganglier med en krok. Den interna halspulsådern är då flamberats med en etsande spets. </li><li> Efter kauterisation, huden är snittet sys, och djuret hålls varm tills helt vaken och återvände sedan till dammen. </li><li> En timme efter ligation är djuret placeras i en sluten kammare infunderas med kväve tills en nivå på 6,0% O 2 nås. Djuret är då utsätts för 35 min av hypoxi. </li><li> Efter hypoxi exponering, återhämtar musen på en värmedyna (33 ° C) i 30 min och sedan tillbaka till dammen. För skapandet av hjärnskada med kombinerad hypoxi / ischemi och infektion / inflammation, är djuret sedan injiceras intraperitonealt med 0,015 ml lipopolysackarid (LPS, 1 mg / kg). </li><li> Fyra dagar post-hypoxia/ischemia, möss bedövas med indirekt kylning på is till den grad att inga reaktioner på skadliga stimuli, sedan perfusion med saltlösning och därefter 4% paraformaldehyd. Hjärnor är bort och cryoprotected. </li></ol><img src="/files/ftp_upload/1951/1951fig1.jpg" alt="Figur 1" /> Figur 1. Karakterisering av hjärnskada i hypoxiska / ischemisk musmodell med H &amp; E färgning. Det finns fokal nekros (pilar) i hjärnans vita substans ipsilaterala (A) till halspulsådern ligering, jämfört med det kontralaterala cerebral vita substansen (B). Ipsilaterala nekros och fokus microinfarcts observeras i hippocampus (C) och Thalamus (D) i hjärnan med ipsilaterala cerebral vita substansen skada. Representativa resultat och siffror: Musmodeller av PVL med hypoxi / ischemi och / eller infektion / inflammation uppnås genom ensidiga halspulsådern ligation följt av exponering för hypoxi och injektion av LPS. Vi definierar den nya musmodeller som lämpliga PVL modeller. Neuropatologiska undersökning av koronalt sektioner färgades med hematoxylin-och eosin (H &amp; E) genom hela framhjärnan avslöjar fokal nekros i centrala vita substansen i hjärnhalva ipsilaterala till halspulsådern ligation (Fig. 1A), med relativt sparsamt av nervceller i den överliggande hjärnbarken jämfört med kontralaterala cerebral vita substansen (Fig. 1B). I fokus nekrotiska vita substansen, är det markant ökning cellularitet bestående av makrofager och reaktiva astrocyter, som är karakteristisk för centrala mänskliga PVL. I den kontralaterala centrala vita substansen är de oligodendroglial kärnor arrangeras i fascicular-liknande paket, denna arkitektur är helt störd av fokal nekros på ipsilaterala sidan. Notera observerar vi också ipsilateralt nekros i hippocampus (bild 1C) och Thalamus (fokala microinfarcts) (Fig. 1D) i hjärnan med ipsilaterala cerebral vita substansen skada. Mänskliga PVL inte heller förekomma i total isolering från medföljande grå lesioner. Det kännetecknande för PVL är den relativa, om än inte fullständig, skona av hjärnbarken överliggande de skadade vita substansen - en funktion som vi tror är närvarande i våra musmodeller. <img src="/files/ftp_upload/1951/1951fig2.jpg" alt="Figur 2" /> Figur 2. Poängsättning skala från 0-5 för att utvärdera vita substansen skada genom MBP eller O1 i musmodeller av periventrikulär leukomalacia. Våra musmodeller visar egenskaper PVL på både fenotypisk och molekylär nivå. Vi har använt immuncytokemiska upptäckt av myelin grundläggande protein (MBP) eller O1-antigen för att utvärdera skador, har etablerat en standard skala från 0 till 5 poäng den vita substansen skada (bild 2), och har präglat den vita substansen patologi (Fig . 3). På senare tid har en icke-partisk stereological analys genomförts för att ersätta den första semi-kvantitativ metod. Dessutom, för att testa för korrelat effekterna av vit substans skada på funktionella resultat har vi utfört beteende tester för motorik. Vi har visat att mössen uppvisar slående motoriken i P10-21 efter hypoxi / ischemi plus LPS på P6. Vi har etablerat poäng kriterier med en skala från 0-4 för att definiera kontralaterala extremiteter dysfunktion skadade möss vid P10-21. Normalt kan normala möss kilar upp en 30 ° lutning (poäng av 4) och klättra upp en 45 ° lutning utan att glida (poäng 3). Skadade möss har lägre poäng i korrelation med graden av vit substans skada. <img src="/files/ftp_upload/1951/1951fig3.jpg" alt="Figur 3" /> Figur 3. Karakterisering av vit substans skada genom MBP eller O1 färgning och myelin struktur genom elektronmikroskop (EM). MBP och O1 färgning visa myelin förlust i cerebrala vit substans ipsilateralt till ligering, jämfört med kontralaterala vita substansen. EM undersökning visar myeliniserade axoner i hjärnans vita substans kontralateral till ligation och degenererade vävnaden i ipsilaterala vit substans. Vi räknar med att inrättandet av den nya musmodeller av PVL ger oss möjlighet att studera specifika mekanismer för PVL-liknande skada i den omogna hjärnan. Vi kommer att behandla patogenes med hjälp av transgena musstammar för att undersöka betydelsen av enskilda molekyler av intresse och utvärdera mål för neuroprotektion.

<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>

Skador på den växande hjärnan leder till förödande neurologiska konsekvenser. Påfallande är mönstret av perinatal hjärnskada starkt åldersberoende. På sikt spädbarn, det påverkar främst hjärnbarken med karakteristiska neuronala förlust. Men i prematura barn, det selektivt påverkar hjärnans vita substans med framstående skada att utveckla oligodendrocyter, kallas en sjukdom periventrikulär leukomalacia (PVL). Omfattningen av problemet med hjärnskada hos prematura barn är extraordinärt. För tidig förlossning och förbättrad neonatal intensivvård har lett till att nästan 90% av överlevnad de 13 miljoner för tidigt födda barn i världen (ca 56 tusen i USA), född årligen med en födelsevikt i 1500 gram, dvs med mycket låg födelsevikt spädbarn. Cirka 10% av de överlevande utställningen därefter cerebral pares och cirka 50% har kognitiva och beteendemässiga underskott. PVL är den viktigaste hjärnan patologi underliggande cerebral pares hos prematura barn. Ingen specifik behandling för PVL finns för närvarande, delvis på grund patogenesen inte har förstått 1,2. Även om etiologin av PVL är multifaktoriell, hypoxi / ischemi med maternell intrauterin infektion perinatal tros vara en primär orsak till PVL. En hypoxiska / ischemisk modell av PVL hos råtta vid P7 har tidigare fastställts 3. Denna modell är unikt genom att det liknar PVL hos prematura barn. Det kännetecknande för denna modell är selektiv vita substansen patologi, i motsats till majoriteten av stroke modeller som kännetecknas av grå hjärnsubstans infarkt. Bör dock begränsningar i hypoxisk-ischemisk modell i råttan inte förbises. För det första är etiologi PVL är ofta multifaktoriell. Förutom att hypoxi / ischemi, moder till foster infektion / inflammation är starkt förknippat med PVL. Således skulle kombinerad hypoxi / ischemi och infektion / inflammation skapar sannolikt mer PVL-liknande modeller. Den gnagare ålder som bäst korrelerar med människans utveckling under största risken för PVL lesioner (24-32 graviditetsveckor) är postnatal eftersom oligodendroglial differentiering är främst en postnatal händelse i gnagare. Eftersom det är omöjligt att producera en moder till foster infektion / inflammation i postnatala ålder, skulle det vara rimligt att simulera en infektion på en relevant postnatal utvecklingsstadium för att skapa en gnagare modell av PVL. Vi har försökt kombinera LPS injektion med hypoxiska / ischemisk modell i möss vid P6, att producera mer kliniskt relevant PVL lesioner. För det andra kräver den optimala proof-of-concept experimentella strategi för en orsakande roll i en molekyl användning av transgena möss, genetiska manipulationer är mycket svårare hos råtta än i musen. Därför har vi försökt att omvandla vår experimentella modell av PVL från råtta till musen. Musmodeller av PVL kommer avsevärt att underlätta 1) studier av sjukdomens biologi och patogenes med hjälp av tillgängliga transgen mus stammar, 2) överledning av läkemedelsprövningar för att upptäcka kandidat agenter med terapeutisk potential, 3) testning av nyttan av stamceller med nedsatt immunförsvar musstammar.

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mouse models of periventricular leukomalacia

Vi beskriver ett protokoll för upprättandet musmodeller av periventrikulär leukomalacia (PVL). PVL är den dominerande formen av hjärnskada hos prematura barn och de vanligaste föregångare av cerebral pares. PVL kännetecknas av periventrikulär vita substansen skadas med framträdande oligodendroglial skada. Hypoxi / ischemi med eller utan systemisk infektion / inflammation är de främsta orsakerna till PVL. Vi använder P6 möss att skapa modeller av neonatal hjärnskada genom induktion av hypoxi / ischemi med eller utan systemisk infektion / inflammation med unilateral halspulsådern ligation följt av exponering för hypoxi, med eller utan injektion av endotoxin lipopolysackarid (LPS). Immunohistokemi av myelin grundläggande protein (MBP) eller O1 och elektron mikroskopisk undersökning visar framstående myelin förlust i cerebrala vit substans med ytterligare skador på hippocampus och thalamus. Inrättande av musmodeller av PVL kommer att kraftigt underlätta studier av sjukdomen patogenes använda tillgängliga transgen mus stammar, överledning av läkemedelsprövningar i en relativt hög genomströmning sätt att identifiera kandidat terapeutiska medel, och testning av stamcellstransplantation med immunbrist musstammar.

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

Mouse Models of Periventricular Leukomalacia

Vi etablerade musmodeller av periventrikulär leukomalacia (PVL), det dominerande hjärnskada hos prematura barn kännetecknas av periventrikulär vita substansen lesioner. Hypoxi / ischemi med / utan systemisk infektion är de främsta orsakerna till PVL. Ensidig halspulsådern ligatur och hypoxi exponering med / utan lipopolysackarid injektion skapar PVL-liknande lesioner i P6 möss.

Musmodeller av periventrikulär Leukomalacia

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

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.

En enkel komposit Fenotyp Scoring System för utvärdering musmodeller av cerebellär ataxi

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

Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions.

We provide a reproducible method for culturing confluent monolayers of human fetal retinal pigment epithelial cells (hfRPE) cells that exhibit morphology, physiology, polarity, and protein and gene expression patterns of adult native tissue. This work has been extended to an animal model of several eye diseases.

Experimentella modeller för studier av näthinnan pigmentepitel fysiologi och patofysiologi

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium ...

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.

Flera-mus neuroanatomiska 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 ...

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.

Olfactory Analyser för musmodeller av neurodegenerativ sjukdom

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.

3D-modellering av de laterala ventriklama och Histologisk karakterisering av periventrikulär Tissue i människor och mus

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).

Använda enzymbaserade biosensorer för att mäta tonic och phasic glutamat i Alzheimers musmodeller

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.

Utforska rymden - avtäckning anatomi periventrikulär strukturer att avslöja de laterala ventriklarna i den mänskliga hjärnan

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.

Om inrättande av musmodeller för Zika Virus-inducerad neurologiska sjukdomar med hjälp av Intracerebral injektion strategier: embryonala, Neonatal och vuxen

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.

Vuxen mus DRG Explant och dissocierade cellmodeller för att undersöka neuroplasticitet och Svaren till miljömässiga förolämpningar inklusive virusinfektion

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 Elektrofysiologisk mätning av sammansatta muskel aktionspotential från frambenen i musmodeller av motorneuronen Degeneration

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