Name: circumscribed capsular infarct modeling using a photothrombotic technique
Uploaded: 2016-06-02T13:00:00
Description: Watch this Scientific Journal Video about Circumscribed Capsular Infarct Modeling Using a Photothrombotic Technique at JoVE.com

This work was supported by a grant from the Institute of Medical System Engineering (iMSE) &#38; GIST-Caltech Collaborative Fund (K04592) from GIST and by the Basic Science Research Program through NRF of Korea funded by the Ministry of Science, ICT and future Planning (NRF-2013R1A2A2A01067890).

All procedures were conducted according to the institutional guidelines of Gwangju Institute of Science and Technology (GIST), and all procedures were approved by the Institutional Animal Care and Use Committee at GIST.
1. Pre-lesioning Steps
<ol>
	<li>Identification of the Forelimb Area in the Internal Capsule using AAV-GFP
	<ol>
		<li>House and handle Sprague Dawley rats (~400 g, 11 - 13 weeks) in compliance with institutional and national guidelines.</li>
		<li>Sterilize all surgical tools an...

<ol>
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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+clinical+importance+of+white+matter+hyperintensities+on+brain+magnetic+resonance+imaging:+systematic+review+and+meta-analysis.">The clinical importance of white matter hyperintensities on brain magnetic resonance imaging: systematic review and meta-analysis.</a> Bmj. 341, 3666 (2010).</li><li>Zhang, K., Sejnowski, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+universal+scaling+law+between+gray+matter+and+white+matter+of+cerebral+cortex.">A universal scaling law between gray matter and white matter of cerebral cortex.</a> PNAS. 97 (10), 5621-5626 (2000).</li><li>Kim, H. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+animal+model+of+capsular+infarct:+endothelin-1+injections+in+the+rat.">An animal model of capsular infarct: endothelin-1 injections in the rat.</a> Behav Brain Res. 169 (2), 206-211 (2006).</li><li>He, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definition+of+the+anterior+choroidal+artery+territory+in+rats+using+intraluminal+occluding+technique.">Definition of the anterior choroidal artery territory in rats using intraluminal occluding technique.</a> J Neurol Sci. 182 (1), 16-28 (2000).</li><li>Tanaka, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+model+of+lacunar+infarction+in+the+gyrencephalic+brain+of+the+miniature+pig:+neurological+assessment+and+histological,+immunohistochemical,+and+physiological+evaluation+of+dynamic+corticospinal+tract+deformation.">Experimental model of lacunar infarction in the gyrencephalic brain of the miniature pig: neurological assessment and histological, immunohistochemical, and physiological evaluation of dynamic corticospinal tract deformation.</a> Stroke. 39 (1), 205-212 (2008).</li><li>Shibata, M., Ohtani, R., Ihara, M., Tomimoto, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=White+matter+lesions+and+glial+activation+in+a+novel+mouse+model+of+chronic+cerebral+hypoperfusion.">White matter lesions and glial activation in a novel mouse model of chronic cerebral hypoperfusion.</a> Stroke. 35 (11), 2598-2603 (2004).</li><li>Watson, B. D., Dietrich, W. D., Busto, R., Wachtel, M. S., Ginsberg, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+reproducible+brain+infarction+by+photochemically+initiated+thrombosis.">Induction of reproducible brain infarction by photochemically initiated thrombosis.</a> Ann Neurol. 17 (5), 497-504 (1985).</li><li>Kuroiwa, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+rat+model+of+photothrombotic+ischemia+and+infarction+within+the+caudoputamen.">Development of a rat model of photothrombotic ischemia and infarction within the caudoputamen.</a> Stroke. 40 (1), 248-253 (2009).</li><li>Bashkatov, A. N., Genina, E. A., Tuchin, V. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+corticospinal+tract+location+at+corona+radiata+and+posterior+limb+of+the+internal+capsule+in+human+brain.">A review of corticospinal tract location at corona radiata and posterior limb of the internal capsule in human brain.</a> NeuroRehabilitation. 24 (3), 279-283 (2009).</li><li>Kim, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+changes+in+resting-state+brain+activity+in+a+capsular+infarct+model.">Longitudinal changes in resting-state brain activity in a capsular infarct model.</a> J Cereb Blood Flow Metab. 35 (1), 11-119 (2014).</li><li>Blasi, F., Whalen, M. J., Ayata, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lasting+pure-motor+deficits+after+focal+posterior+internal+capsule+white-matter+infarcts+in+rats.">Lasting pure-motor deficits after focal posterior internal capsule white-matter infarcts in rats.</a> J Cereb Blood Flow Metab. 35 (6), 977-984 (2015).</li><li>Metz, G. A., Antonow-Schlorke, I., Witte, O. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+improvements+after+focal+cortical+ischemia+in+adult+rats+are+mediated+by+compensatory+mechanisms.">Motor improvements after focal cortical ischemia in adult rats are mediated by compensatory mechanisms.</a> Behavioural brain research. 162 (1), 71-82 (2005).</li></ol>

The capsular infarct model presented here demonstrates a targeted lesion with marked and persistent motor impairment in forelimb function. Previous models of subcortical capsular stroke have demonstrated an insufficient degree of motor impairment and a rapid recovery process6,8,9 . In this sense, this model resembles the clinical capsular infarct cases which exhibit long-term functional impairment.
The most critical steps in the development of a circumscribed capsular infarct model ...

Until recently, the &#34;gray matter stroke (GMS) models&#34; have been exclusively used to understand the pathophysiology of stroke and to guide the development of new treatments. However, there has been an increasing prevalence of stroke that affects subcortical white matter in elderly individuals, which constitutes 15 - 25% of all strokes1,2. Numerous studies have been conducted regarding stroke using GMS models, whereas there are few studies that have used white matter stroke (WMS) models. White matter in rodents is substantially less than the white matter in humans or primates. Consequently, it is more difficult to selectively access and destroy the target regions in the white matter3. Additionally, no efficient tools have been developed to date to selectively destroy the planned extent of targeted white matter. Therefore, there has been lack of appropriate models for study of white matter strokes.
Animal stroke models are often used to monitor the progress of motor recovery for development of new rehabilitative and therapeutic methods. It is ideal to utilize an animal model that exhibits a long-term neurological deficit concordant with the anatomical alterations demonstrated in human stroke4,5. In this regard, rapid recovery of the motor deficit and wide involvement of the brain following infarct lesioning may not be realistic in the pursuit of stroke research. Previous capsular infarct models have been made by the occlusion of the internal carotid or anterior choroidal arteries and diffusion of endothelin-1 (ET-1) into the internal capsule6-9. Nonetheless, artery occlusion requires careful dissection of arteries, but it produces a wide area of infarct lesion, including the internal capsule, without persistent behavioral deficits. Moreover, ET-1 was not diffused to completely destroy the posterior limb of the internal capsule, and hence less marked or persisting behavioral deficit.
A photothrombotic infarction model has been widely used to generate various types of infarct lesions in the cortex and subcortical structures10. The technique include intravenous administration followed by focal illumination, which leads to platelet aggregation in the small vessels and generation of infarct lesions10. Photothrombotic technique has been extensively used to create GMS lesions, whereas it has rarely been used to generate WMS lesions5,11. For this technique, a combination of Rose Bengal dye and light irradiation has been demonstrated to be useful in destruction of the target structure, causing corresponding functional deficits. The key element of the photothrombotic technique is light irradiation, because it determines the size of infarct lesions. Light irradiation results in different effects on gray matter and white matter, because the scattering of light is more than 4 times higher in white matter compared with gray matter12; Accordingly, if the light intensity has a sufficiently low irradiance (&#60;1,140 mW/mm2), one can limit the extension to which photothrombotic lesion affect the extent to the white matter (i.e., internal capsule). For example, light of higher energy can induce infarcts in both gray and white matter, yet lower energy light may induce photothrombosis only in white matter. Furthermore, the penetration of light energy was very limited. Approximately 99% of light energy was lost beyond 1 mm from the source of light13. Therefore, it is expected that accurately targeted, lower energy light induces photothrombosis only in the white matter with a minimal encroachment of the neighboring gray matter.
Here, we describe a novel method to create infarct lesions in the forelimb area of the internal capsule in rodents. We describe the method of identification of the forelimb area in the internal capsule, the technology of light irradiation, including the adjustment and delivery of light, and the generation of an infarct lesion. We also describe behavioral testing used to evaluate the completeness of the capsular modeling.

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circumscribed capsular infarct modeling using a photothrombotic technique

Recent increase in the prevalence rate of white matter stroke demands specific research in the field. However, the lack of a pertinent animal model for white matter stroke has hampered research investigations. Here, we describe a novel method for creating a circumscribed capsular infarct that minimizes damage to neighboring gray matter structures. We used pre-surgery neural tracing with adeno-associated virus-green fluorescent protein (AAV-GFP) to identify somatotopic organization of the forelimb area within the internal capsule. The adjustment of light intensity based on different optical properties of gray and white matter contributes to selective destruction of white matter with relative preservation of gray matter. Accurate positioning of optical-neural interface enables destruction of entire forelimb area in the internal capsule, which leads to a marked and persistent motor deficit. Thus, this technique produces highly replicable capsular infarct lesions with a persistent motor deficit. The model will be helpful not only to study white matter stroke (WMS) at the behavioral, circuit, and cellular levels, but also to assess its usefulness for development of new therapeutic and rehabilitative interventions.

&#160; The method presented here is intended to create a circumscribed capsular infarct with a persistent motor deficit. Therefore, it is critical to correctly determine the target within the internal capsule in the pre-surgery step. The somatotopic mapping of pyramidal fibers in the internal capsule has not been settled to date. To correctly identify the target within the internal capsule, the forelimb area must be delineated. An injection of AAV-GFP into the forelimb area of the motor c...

Watch this Scientific Journal Video about Circumscribed Capsular Infarct Modeling Using a Photothrombotic Technique at JoVE.com

Circumscribed Capsular Infarct Modeling Using a Photothrombotic Technique

This manuscript describes a modeling technique of capsular infarct. Here we utilized a modified photothrombotic technique with low intensity of light after pre-surgery target mapping. Using this technique, we created a circumscribed capsular infarct model with persistent motor impairment.

Recent increase in the prevalence rate of white matter stroke demands specific research in the field. However, the lack of a pertinent animal model for ...

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