Bu çalışma, Ulusal Sağlık Enstitüleri (RO1 NS059043 ve RO1 ES015988), Ulusal Multipl Skleroz Derneği, Anemi Araştırma, Feldstein Tıp Vakfı Roche Vakfı ve Çocuk Hastaneleri Shriners hibe tarafından desteklenen parçası. Biz onun önerileri için Dr David Pleasure teşekkür etmek istiyorum. Bu çalışma kısmen, Ulusal Sağlık Enstitüleri WD hibeler (RO1 NS059043 ve RO1 ES015988), Ulusal Multipl Skleroz Derneği, Çocuklar için Shriners Hastaneler tarafından desteklenmiştir. Biz bu makalede ilgili hiçbir çatışan çıkarları beyan ederim.

P6 farelerde neonatal beyin hasarı oluşturarak detaylı prosedürler: <ol><li> Postnatal günde 6 (P6) fareler zararlı stimülasyonu (derin anestezi) yanıtsızlık noktasında buz üzerinde dolaylı soğutma ile anestezi. </li><li> Alkol ile cilt temizliği takiben anterior boyun, karın orta hat kesi yapılır. </li><li> Diseksiyon mikroskobu altında, sağ karotis arter bifurkasyon nazikçe geri çekilmeden omohyoid ve sternokleidomastoid kaslar tarafından yaklaştı. </li><li> Carotica&#39;nın etrafında fasya kaldırılır ve yakındaki bir kanca ile vagus siniri ve sempatik ganglionlar izole proksimal iç dalı. Internal karotid arter sonra dağlama ucunu kullanarak koterize. </li><li> Koterizasyon ardından, cilt insizyonu sütüre edilir ve daha sonra tamamen uyanık ve baraj döndü kadar hayvan sıcak tutulur. </li><li> Ligasyonu Bir saat sonra, hayvan, O 2 ulaşıldığında% 6.0 seviyesine kadar azot ile aşılanmış yalıtılmış bir bölmede yer alır . Hayvan hipoksi 35 dakika sonra maruz kalmaktadır. </li><li> Hipoksiye maruz kaldıktan sonra, farenin 30 dakika için bir ısıtma yastığı (33 ° C) kurtarır ve daha sonra baraj iade edilir. Kombine hipoksi / iskemi ve enfeksiyon / enflamasyon ile beyin hasarı oluşturulması için, hayvanın daha sonra 0.015 ml lipopolisakkarid (LPS, 1 mg / kg) intraperitoneal olarak enjekte edilir. </li><li> Post-hypoxia/ischemia Dört gün sonra sonra tuzlu su ve% 4 paraformaldehid ile perfüze zararlı uyaranlara yanıtsızlık noktasında buz üzerinde dolaylı soğutma, fareler anestezi. Brain kaldırılır ve cryoprotected. </li></ol><img src="/files/ftp_upload/1951/1951fig1.jpg" alt="Şekil 1" /> Şekil 1 / hipoksik iskemik fare modelinde H &amp; E boyama ile beyin hasarı Karakterizasyonu. (A) karotis ligasyonu ipsilateral serebral beyaz cevherde fokal nekroz (oklar) kontralateral serebral beyaz madde (B) ile karşılaştırıldığında vardır. Ipsilateral nekroz ve odak microinfarcts hipokampus (C) ve ipsilateral serebral beyaz cevher hasarı ile beyin talamus (D) görülmektedir. Temsilcisi Sonuçlar ve Şekiller: Hipoksi / iskemi ve / veya enfeksiyon / enflamasyon PVL fare modelleri, hipoksi ve LPS enjeksiyonu maruz kalma ardından tek taraflı karotis ligasyonu ile elde edilir. Biz yeni fare modelleri uygun PVL model olarak tanımlıyoruz. Nöronların göreceli koruyucu tüm önbeyin üzerinden hematoksilen-eozin (H &amp; E) ile boyandı koronal bölümden nöropatolojik inceleme ile karotis ligasyonu (Şekil 1A) ipsilateral serebral hemisferin merkezi beyaz cevherde fokal nekroz ortaya örten serebral korteksin kontralateral serebral beyaz cevher (Şekil 1B). Fokal nekrotik beyaz cevher, odak insan PVL karakteristik olarak makrofaj ve reaktif astrositler hücresellik oluşan artış var işaretlenir. Kontralateral merkezi beyaz cevher, oligodendroglial çekirdekleri fasiküler gibi demetleri düzenlenmiş; bu mimari, ipsilateral tarafta fokal nekroz tamamen bozulur. Unutmayın ki, biz de ipsilateral serebral beyaz cevher hasarı ile beyinlerinde ipsilateral hipokampus nekroz (Şekil 1C) ve talamus (odak microinfarcts) (Şekil 1D) gözlemlemek. İnsan PVL da gri madde lezyonları ile birlikte komple izolasyon oluşmaz. Fare modellerinde mevcut olduğunu düşündüğünüz bir özelliği - PVK işaretidir yaralı beyaz cevher örten serebral korteksin koruyucu, tam akrabası, olmasa da. <img src="/files/ftp_upload/1951/1951fig2.jpg" alt="Şekil 2" /> Şekil 2 MBP veya O1 periventriküler lökomalazi fare modellerinde beyaz cevher hasarı değerlendirmek için 0-5 Puanlama ölçekli. Bizim fare modelleri, fenotipik ve moleküler düzeyde hem de PVL özellikleri göstermektedir. Biz miyelin bazik protein (MBP) veya A1-antijen immünositokimyasal algılama, yaralanma değerlendirmek için kullanmışlardır beyaz cevher hasarı (Şekil 2) puan, 0 ila 5 standart bir ölçek kurduk ve beyaz cevher patolojisi (Şekil karakterize var 3). Daha yakın zamanlarda, önyargılı olmayan bir stereolojik tahlil, ilk yarı-kantitatif bir yöntem yerine uygulamaya konmuştur. Buna ek olarak, fonksiyonel sonuçlar üzerine beyaz cevher hasarı bağıntılı etkileri test etmek için, motor fonksiyon için davranışsal testler gerçekleştirdik. Biz farelerin P6 az aşağıdaki P10-21 hipoksi / iskemi artı LPS çarpıcı motor defisiti sergilediklerini göstermiştir. P10 yaralı farelerde kontrlateral ekstremite fonksiyon bozukluğu tanımlamak için bir ölçekte 0-4 puanlama kriterleri kurduk-21. Tipik olarak, 30 ° eğim (4 puan) normal fareler kadar koşar ve 45 kadar tırmanmaya ° sürgülü olmadan eğim (3 puan). Yaralı fareler beyaz cevher hasarı derecesi ile korelasyon daha düşük puanlar var. <img src="/files/ftp_upload/1951/1951fig3.jpg" alt="Şekil 3" /> Şekil 3 MBP veya O1 boyama ve miyelin yapısı elektron mikroskobu (EM) ile beyaz cevher hasarı Karakterizasyonu. MBP ve kontralateral beyaz maddeye göre ligasyonu ipsilateral serebral beyaz cevher O1 boyama gösterisi miyelin kaybı,. EM muayene ligasyonu kontralateral serebral beyaz cevher miyelinli aksonlar gösterir ve ipsilateral beyaz cevher doku dejenere. Biz PVL yeni fare modelleri kurulması PVL gibi yaralanma özel mekanizmalar bize immatür beyin çalışma izin olacağını tahmin ediyoruz. Biz bireysel ilgi moleküllerin rolünü incelemek ve nöro hedefleri değerlendirmek için transgenik fare suşu kullanılarak patogenezinde ele alınacaktır.

<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>

Gelişmekte olan beyin yaralanması, yıkıcı nörolojik sonuçlara yol açar. Çarpıcı, perinatal beyin hasarı desen yüksek oranda yaşa bağlıdır. Term bebeklerde, ağırlıklı olarak, karakteristik nöron kaybı ile serebral korteks etkiler. Ancak, prematüre bebeklerde, seçici, gelişmekte olan oligodendrosit önde gelen yaralanma ile serebral beyaz cevher etkiler periventriküler lökomalazi (PVL) olarak adlandırılan bir hastalık. Prematüre bebekte beyin hasarı sorunun büyüklüğü olağanüstü. Erken doğum ve gelişmiş yenidoğan yoğun bakım yıllık bir doğum ağırlığı 1500 gr altında, yani çok düşük doğum ağırlıklı bebeklerde doğmuş dünya çapında 13 milyon prematüre bebeklerde (Amerika Birleşik Devletleri yaklaşık 56.000) yaklaşık% 90 hayatta kalma yol açmıştır. Kurtulanların yaklaşık% 10 daha sonra serebral palsi sergiler ve yaklaşık% 50 oranında bilişsel ve davranışsal eksiklikler var. PVL prematüre bebeklerde en önemli beyin altta yatan patolojinin serebral palsi. Patogenezinde 1,2 iyi anlaşılmaktadır değil, çünkü PVL için spesifik bir tedavisi yok şu anda kısmen var. PVL, etyolojisi multifaktöriyel olmasına rağmen, anne rahim içi enfeksiyonu olan perinatal hipoksi / iskemi PVL bir birincil nedeni olduğu düşünülmektedir. P7 sıçan PVL / A hipoksik iskemik modeli daha önce 3 kurulmuştur. Bu model, prematüre bebeklerde PVL benzediği eşsizdir. Bu modelin en belirgin özelliği, gri cevherde enfarktüsü ile karakterize olan inme modelleri çoğunluğunun aksine, seçici beyaz madde patolojisi. Ancak, sıçanlarda hipoksik-iskemik model sınırlamaları göz ardı edilmemelidir. Öncelikle, PVL etiyolojisinde genellikle multifaktöryel. Hipoksi / iskemi, maternal-fetal enfeksiyon / inflamasyona ek olarak güçlü bir PVL ile ilişkilidir. Böylece, kombine hipoksi / iskemi ve enfeksiyon / enflamasyon büyük olasılıkla daha PVL gibi modeller oluşturmak. Oligodendroglial farklılaşma, ağırlıklı olarak bir doğum sonrası olay kemirgenler, çünkü en iyi PVL lezyonlar (gebeliğin 24-32 hafta) için en büyük risk, insan gelişim dönemi ile ilişkilidir kemirgen yaş postnatal. Doğum sonrası yaşlarda anne-fetal enfeksiyon / iltihap üretmek olanaksız olduğundan, PVL bir kemirgen modeli oluşturmak için ilgili bir postnatal gelişim aşamasında bir enfeksiyon simüle etmek mantıklı olacaktır. Biz klinik olarak daha anlamlı PVL lezyonlar üretmek için, LPS enjeksiyonu P6 az farelerde / hipoksik iskemik modeli ile birleştirmek için çalışmışlardır. İkincisi, bir molekülün etken bir rol için en uygun proof-of-concept deneysel yaklaşım, transgenik farelerin kullanımı gerektirmektedir; genetik manipülasyonlar sıçan fare göre çok daha zor. Böylece, sıçan, fare PVL bizim deneysel model dönüştürmek için çalışmışlardır. 2) tedavi potansiyel aday ajanları keşfetmek için ilaç denemelerinin iletimi; 3) immünyetmezligi fare suşu kullanılarak kök hücre yardımcı programı test PVL Fare modelleri 1) transgenik fare suşu kullanılarak hastalığın biyolojisi ve patogenezi çalışma büyük ölçüde kolaylaştıracaktır.

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mouse models of periventricular leukomalacia

Biz periventriküler lökomalazi (PVL) fare modelleri kurmak için bir protokol tanımlamaktadır. PVL, beyin hasarı, prematüre bebeklerde serebral palsi en yaygın öncül baskın şeklidir. PVL önde gelen oligodendroglial yaralanması olan periventriküler beyaz cevher hasarı ile karakterizedir. Sistemik enfeksiyon / enflamasyon ya da hipoksi / iskemi PVL birincil nedenleri. P6 fareler iskemi / hipoksi indüksiyon veya maruz kalma ya da hipoksi, endotoksin lipopolisakkarid (LPS) enjeksiyonu olmadan tek taraflı karotis ligasyonu ile sistemik enfeksiyon / iltihap olmadan yenidoğan beyin hasarı modeller yaratmak için kullanın. Miyelin bazik protein (MBP) veya O1 ve elektron mikroskobik inceleme İmmünohistokimya, hipokampus ve talamus ek hasar serebral beyaz cevherde belirgin miyelin kaybı göstermektedir. PVL fare modelleri Kuruluş hastalığının patogenezinde immün yetmezlik fare suşu kullanılarak transgenik fare suşları, ilaç denemelerinin terapötik ajanlar aday belirlemek için nispeten yüksek kapasiteli bir şekilde iletimi, ve kök hücre nakli testi kullanılarak yapılan çalışmada büyük ölçüde kolaylaştıracaktır.

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

Mouse Models of Periventricular Leukomalacia

Biz periventriküler lökomalazi (PVL), prematüre bebeklerde periventriküler beyaz cevher lezyonları ile karakterize baskın beyin hasarı fare modelleri kurdu. Hipoksi / iskemi / sistemik enfeksiyon olmadan PVL birincil nedenleri. Tek taraflı karotis ligasyonu ve / lipopolisakkarid enjeksiyon olmadan hipoksi maruz P6 fareler PVL benzeri lezyonlar oluşturur.

Periventriküler lökomalazi Fare Modelleri

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 Görüntüleme.  University of California, Davis. Biz periventriküler lökomalazi (PVL), prematüre bebeklerde periventriküler beyaz cevher lezyonları ile karakterize baskın beyin hasarı fare modelleri kurdu. Hipoksi / iskemi / sistemik enfeksiyon olmadan PVL birincil nedenleri. Tek taraflı karotis ligasyonu ve / lipopolisakkarid enjeksiyon olmadan hipoksi maruz P6 fareler PVL benzeri lezyonlar oluşturur.

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.

Serebellar Ataksi Fare Modelleri Değerlendirme Basit Kompozit Fenotip Puanlama Sistemi

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

Nörobilim

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.

Retina Pigment Epitel Fizyoloji ve Patofizyoloji Çalışma Deneysel Modeller

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.

Çoklu-fare nöroanatomik Manyetik Rezonans Görüntüleme

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.

Nörodejeneratif Hastalığının fare modelleri için koku alma Tahliller

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.

İnsanlar ve Mouse Periventriküler Doku Yanal ventrikülleri ve histolojik Karakterizasyonu 3D Modelleme

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.

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).

Enzim bazlı biyosensörlerin kullanılması Alzheimer Fare Modellerinde Tonik ve Fazik glutamat Ölçmek için

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.

Derin uzay - anatomi Lateral ventrikül insan beyninin ortaya çıkarmak için periventriküler yapıların ortaya çıkararak keşfetmek

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.

İntraserebral enjeksiyon stratejileri kullanarak Zika virüs kaynaklı nörolojik bozukluklar için fare modelleri oluşturma: embriyonik, Yenidoğan ve yetişkin

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.

Yetişkin fare DRG Explant ve nöroplastisite ve Viral enfeksiyon da dahil olmak üzere çevre hakaret yanıtlar araştırmak için hücre modelleri ayrışmış

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.

Vivo Bileşik kas aksiyon potansiyeli Motor nöron hastalığı, fare modellerinde ön ayakları üzerinden elektrofizyolojik ölçümü

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