Name: triggering reactive gliosis in vivo by a forebrain stab injury
Uploaded: 2015-06-29T13:00:00
Description: Watch this Scientific Journal Video about Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury at JoVE.com

We thank Katherine Clark for technical assistance. A.D.R.G. is funded in part by 5K01MH097957-03

واستخدمت الكبار (3-4 أشهر من العمر) ذكور الفئران على مختلطة C57BL / 6 الخلفية في هذا البروتوكول. تم الاحتفاظ الحيوانات على 12 ساعة ضوء / دورة الظلام، وسمح حرية الوصول إلى الغذاء والماء. أجريت جميع الإجراءات المتبعة في هذا البروتوكول وفقا لبروتوكولات وافقت عليها اللجنة دريك?...

<ol>
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فمن الأهمية بمكان أن الجمجمة أو الجافية الأساسية لا تلف أثناء الحفر. استخدام الضغط الخفيف أثناء الحفر لضمان الجمجمة لم يتم ثقب. بالإضافة إلى ذلك، يجب توخي الحذر في حين رفع قطعة الجمجمة لضمان الجافية لا انطلق مع العظم. إصابة الدما?...

A major challenge for developing successful therapies to treat the injured CNS is an incomplete understanding of the complex multicellular events that are triggered by the trauma. Reactive astrocytes are gaining increasing recognition as a promising target for novel therapies1. Though historically regarded as hostile to neural repair, reactive astrocytes are now recognized as critical components of a complex, multicellular neuroprotective response that includes attenuation of inflammatory processes and limiting secondary damage and neurodegeneration2-6. Although the neuropathological characteristics of reactive gliosis have long been well defined, the cellular and molecular mechanisms regulating reactive gliosis, and the diverse array of downstream consequences remain poorly understood. Understanding the mechanisms that drive reactive gliosis, as well as the subsequent cellular and molecular events, is an important step towards developing strategies aimed at promoting the neuroprotective properties of reactive gliosis, while attenuating the detrimental effects. 
Here we demonstrate a method to induce severe reactive astrogliosis in the forebrain of adult mice using a stab injury. In contrast to other traumatic brain injury (TBI) models, such as controlled cortical impact (CCI) or fluid percussion injury (FPI), which require specialized equipment to produce an injury, the forebrain stab requires only a stereotaxic device to stabilize the head and a No. 11 scalpel blade. Thus the forebrain stab lesion model is more broadly accessible to a wide range of laboratories that do not have access to the specialized devices necessary for creating an FPI or CCI injury. The method described here enables investigators to reliably and reproducibly trigger a robust gliosis response to investigate subsequent cellular and molecular events. Once recovered from surgery, animals that have received a forebrain stab injury can survive for prolonged periods without the need for specialized care and can be returned to the colony for acute, intermediate, or chronic studies. Though less clinically translatable than FPI or CCI models of TBI, a forebrain lesion produced by a stab injury serves as a simple yet useful experimental model to investigate basic biological mechanisms underlying reactive gliosis and other neuropathological events following trauma to the CNS.

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			<td>High speed micro drill</td>
			<td>Harvard Apparatus</td>
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			<td>stainless steel scalpel blade, #11</td>
			<td>MedVet</td>
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			<td>5/45 angled forceps</td>
			<td>Fine Science Tools</td>
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			<td>Gelfoam sponge 12cmx7mm</td>
			<td>Fisher</td>
			<td>NC9841478</td>
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			<td>Antibodies and other reagents</td>
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			Dilution (fluorescence)</td>
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			<td>Rb anti-GFAP</td>
			<td>DAKO</td>
			<td>&#160;Z033429-2</td>
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			<td>Shp anti-BrdU</td>
			<td>Abcam</td>
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			1:500</td>
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			<td>Biotinylated goat anti-rabbit</td>
			<td>Vector Laboratories</td>
			<td>BA-1000&#160;</td>
			<td>1:400</td>
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			<td>Biotinylated rabbit anti-sheep</td>
			<td>Vector Laboratories</td>
			<td>BA-6000</td>
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			<td>Alexafluor 488 goat anti-rabbit</td>
			<td>Life Technologies</td>
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			<td>Alexafluor 568 donkey anti-sheep</td>
			<td>Life Technologies</td>
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			<td>DAPI Nucleic Acid Stain</td>
			<td>Life Technologies</td>
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			<td>1:1000</td>
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			<td>Cresyl Violet Acetate</td>
			<td>Sigma Aldrich</td>
			<td>C5042-10G</td>
			<td>1%</td>
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triggering reactive gliosis in vivo by a forebrain stab injury

Following injury to the CNS, astrocytes undergo a broad range of biochemical, morphological, and molecular changes collectively referred to as reactive astrogliosis. Reactive astrocytes exert both inflammatory and protective effects that inhibit and promote, respectively, neural repair. The mechanisms underlying the diverse functional properties of reactive astrogliosis are not well understood. Achieving a greater understanding of these mechanisms is critical to developing therapeutic strategies to treat the injured CNS. Here we demonstrate a method to trigger reactive astrogliosis in the adult mouse forebrain using a forebrain stab lesion. This lesion model is simple, reliable, and requires only a stereotaxic device and a scalpel blade to produce the injury. The use of stab lesions as an injury model in the forebrain is well established and amenable to studies addressing a broad range of neuropathological outcomes, such as neuronal degeneration, neuroinflammation, and disruptions in the blood brain barrier (BBB). Thus, the forebrain stab injury model serves as a powerful tool that can be applied for a broad range of studies on the CNS response to trauma.

لأن الحيوانات تمر هذا الإجراء لا يتطلب تدرج فترات بقاء المتخصصة الرعاية اللاحقة للعمليات الجراحية، قصيرة أو طويلة الأجل بسهولة في الدراسة، وهذا يتوقف على الحاجة إلى تحقيق أمراض حادة أو مزمنة بعد الاصابة. الملامح الرئيسية لدباق رد الفعل، مثل upregulation من GFAP وتضخم في سو?...

Watch this Scientific Journal Video about Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury at JoVE.com

Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury

This article describes a detailed protocol to produce a forebrain stab injury in adult mice. The stab injury induces severe reactive gliosis and glial scar formation which can be subsequently examined by standard immunohistochemistry methods.

مما اثار رد الفعل دباق<em&gt; في فيفو</em&gt; من الدماغ الأمامي الطعنة الإصابات

Following injury to the CNS, astrocytes undergo a broad range of biochemical, morphological, and molecular changes collectively referred to as reactive ...

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Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

و في الجسم الحي النموذجي القوارض من الإصابات الناجمة عن انكماش ، وغير الغازية ورصد الانتعاش

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

زرع الشريان السباتي الكفة لتحريك القص من التوتر في الفئران المستحثة تصلب الشرايين

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

In vivo 19F MRI for Cell Tracking

In vivo 19F MRI for Cell Tracking

في الجسم الحي 19 F التصوير بالرنين المغناطيسي لتتبع الخليوي

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

القياس عن بعد داخل الرحم لقياس ماوس مقلص الضغط<em&gt; في فيفو</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

الكشف عن القيامة في فيفو قبل بيوسينسور كاسباسيتراكير

Anastasis (Greek for &#8220;rising to life&#8221;) is a recently discovered cell recovery phenomenon whereby dying cells can reverse late-stage cell death ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

نضوب خلايا الفأر من Xenografts ورم الإنسان يحسن بشكل ملحوظ تحليل المصب من الخلايا المستهدفة

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

A Repetitive Concussive Head Injury Model in Mice

والمتكرر الارتجاج إصابات الرأس النموذجي في الفئران

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

فتح القصبة الهوائية المعدة حمض الطموح فأري موديل الحاد الرئة إصابة النتائج في القصوى الحاد غير قاتل الرئة الإصابات

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

استنساخه الشرياني التعرية الإصابات وذلك Infrarenal البطن الأبهري لقط في نموذج الفئران

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

نموذج إصابة المورفولوجية في فيفو لدراسة نيوروريجينيريشن في الطرفية والجهاز العصبي المركزي

The regrowth capacity of damaged neurons governs neuroregeneration and functional recovery after nervous system trauma. Over the past few decades, various ...