Name: a preclinical model to assess brain recovery after acute stroke in rats
Uploaded: 2019-11-06T13:00:00
Description: Watch this Scientific Journal Video about A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats at JoVE.com

This work was supported by the National Natural Science Foundation of China (81603315, 81603316), Key R &#38; D plan of Jiangxi province in China (20171ACH80001), Industrial and academic cooperation projects in colleges and universities of Fujian province in China (2018Y41010011).

The procedure and use of animal subjects have been approved by the National Institute of Health for the care and use of laboratory animals. This protocol is specifically adjusted for the tests of middle cerebral artery occlusion/reperfusion (MCAO/R) and sensorimotor function.
1. Experimental design and grouping
<ol>
	<li>Use a rat MCAO/R model to screen a rat brain ischemic model method with more severe brain injury and greater model success ratio using Longa&#39;s score and TTC staining.
	<...

<ol>
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Many models establishing methods and behavioral indicators that are well used in acute cerebral ischemia may not have significant changes in the recovery and sequela stages of brain ischemia16,17. However, the number of patients with brain ischemic in the recovery and sequela stages is the greatest. It is essential to select a suitable animal model for the recovery and sequela stages of ischemia stroke.
We use the MCAO/R model in rats ...

Brain ischemia is divided into three stages with different post-stroke indicators: the acute stage (within 1 week), the recovery stage (1 week to 6 months), and the sequelae stage (more than 6 months). Presently, most studies focus on the acute stage of brain ischemia because of its significant effect and multi-relative research models1,2,3. However, the recovery and sequelae stages of brain ischemia cannot be ignored due to their long-term complication of disabilities. Therefore, the purpose of this study is to explore a stable, reliable and relatively simple animal model to research the recovery and sequela stages of brain ischemia.
Among the many experimental brain ischemia models, we use middle cerebral artery occlusion (MCAO) via thread bolt insertion into the right middle cerebral artery (MCA). This model is similar to human stroke, which can produce larger infarct volumes, result in many behavioral disorders related to stroke, and can allow blood reperfusion (R) by removing the thread bolt4,5,6. MCAO/R is also considered the gold standard animal model of brain ischemia7. Furthermore, the severity of the brain injury depends on the diameter and the insertion length of the thread bolt, the duration of brain ischemia, and the animal weight (larger rats have bigger brains and thicker cerebral vessels)8. Therefore, by changing the thread bolt type, the infarct time, and the rat weight, a suitable model can be found for the recovery and sequela stages of brain ischemia in MCAO/R rats. To validate the rat model, we performed a 1-day, 35-day, 60-day, and 90-day study of the MCAO/R model using TTC staining and sensorimotor function experiments (a bilateral asymmetry test, a grid-walking test, a rotarod test and a lifting rope test).

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anatomical Microscope</td>
			<td>Leica (Germany)</td>
			<td>S8</td>
			<td>Microscopic operating instrument</td>
		</tr>
		<tr>
			<td>Blade</td>
			<td>Gellette</td>
			<td>/</td>
			<td>Cutting brain sections</td>
		</tr>
		<tr>
			<td>Constant Temperature Shaking Bed</td>
			<td>Taicang Experimental Equipment Factory</td>
			<td>THZ-C</td>
			<td>To keep the brain sections stained evenly and at a constant temperature</td>
		</tr>
		<tr>
			<td>Digital Camera</td>
			<td>Canon</td>
			<td>700D</td>
			<td>For taking pictures of TTC staining</td>
		</tr>
		<tr>
			<td>Electric Shaver</td>
			<td>Shanghai Yuyan Scientific Instruments Co., Ltd.</td>
			<td>3000#</td>
			<td>Removal of hair from the neck of rats</td>
		</tr>
		<tr>
			<td>Forceps Hamostatic</td>
			<td>Shanghai Medical device Co., Ltd.</td>
			<td>14 cm</td>
			<td>Using for brain removing</td>
		</tr>
		<tr>
			<td>Image Pro Plus Software</td>
			<td>Media Cybernetics Inc.</td>
			<td>6.0</td>
			<td>Analyze the infarct volume</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science</td>
			<td>217170702</td>
			<td>Anesthetic gas</td>
		</tr>
		<tr>
			<td>Microforceps</td>
			<td>Shanghai Jinzhong Medical Devices Co., Ltd.</td>
			<td>10 cm</td>
			<td>Vascular micromanipulation</td>
		</tr>
		<tr>
			<td>Microshear</td>
			<td>Shanghai Jinzhong Medical Devices Co., Ltd.</td>
			<td>10 cm</td>
			<td>Vascular micromanipulation</td>
		</tr>
		<tr>
			<td>Ophthalmic Forceps</td>
			<td>Shanghai Jinzhong Medical Devices Co., Ltd.</td>
			<td>10 cm</td>
			<td>Auxiliary skin and muscle anatomy</td>
		</tr>
		<tr>
			<td>Pphthalmic Scissors</td>
			<td>Shanghai Jinzhong Medical Devices Co., Ltd.</td>
			<td>10 cm</td>
			<td>Using for cutting the skin of neck</td>
		</tr>
		<tr>
			<td>Rat Brain Slice Mold</td>
			<td>Shanghai Yuyan Scientific Instruments Co., Ltd.</td>
			<td>400 g</td>
			<td>For standard, uniform cutting of brain tissue</td>
		</tr>
		<tr>
			<td>Rat Rotating Bar Fatigue Apparatus</td>
			<td>Anhui Zhenghua Biological Instrument and Equipment Co., Ltd.</td>
			<td>ZH-300B</td>
			<td>To test the sensorimotor function</td>
		</tr>
		<tr>
			<td>Small Animal Anaesthesia Machine</td>
			<td>Shanghai Yuyan Scientific Instruments Co., Ltd.</td>
			<td>ABM3000</td>
			<td>A gas anesthetic machine</td>
		</tr>
		<tr>
			<td>Small Animal Thermostat</td>
			<td>Beijing Damida Technology Co., Ltd.</td>
			<td>DM.7-YLS-20A</td>
			<td>To maintain animal body temperature constant during operation</td>
		</tr>
		<tr>
			<td>Surgical Scissors</td>
			<td>Shanghai Medical device Co., Ltd.</td>
			<td>16 cm</td>
			<td>Using for decapitate and brain removing</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Shanghai Jinhuan Medical Devices Co., Ltd.</td>
			<td>4-0 / 5-0</td>
			<td>Using for skin and muscle sutures / Using for vascular ligations</td>
		</tr>
		<tr>
			<td>Thread Bolt</td>
			<td>Beijing Cinontech Co. Ltd.</td>
			<td>2636/2838/3040/3043-A4</td>
			<td>Blockage of the middle cerebral artery in rats</td>
		</tr>
		<tr>
			<td>5-triphenyl-2H-tetrazolium chloride (TTC)</td>
			<td>Sigma</td>
			<td>LOT#BCBP3272V</td>
			<td>Brain section staining reagent</td>
		</tr>
	</tbody>
</table>

a preclinical model to assess brain recovery after acute stroke in rats

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery occlusion/reperfusion (MCAO/R) model in male Sprague-Dawley rats was chosen. By changing the rat&#39;s weight (260&#8722;330 g), the thread bolt type (2636/2838/3040/3043) and the brain infarct time (2-3 h), a higher Longa&#39;s score, a larger infarct volume and a greater model success ratio were screened using the Longa&#39;s score and TTC staining. The optimum model condition (300 g, 3040 thread bolt, 3 h brain infarct time) was acquired and used in a 1-90 day observation period after reperfusion via assessment of sensorimotor functions and infarct volume. At these conditions, the bilateral asymmetry test had a significant difference from 1 to 90 days, and the grid-walking test had a significant difference from 1 to 60 days; both differences could be a suitable sensorimotor functional test. Thus, the most appropriate condition of a novel rat model in the recovery and sequela stages of brain ischemia was found: 300 g rats that underwent MCAO with a 3040 thread bolt for a 3 h brain infarct and then reperfused. The appropriate sensorimotor functional tests were a bilateral asymmetry test and a grid-walking test.

Using the abovementioned procedure for a MCAO/R model with a Longa&#39;s score and TTC staining, different treatments of average weight (275/300/320 g), bolt types (2636/2838/3040/3043; Table 1) and ischemic times (2-3 h) and 1 day reperfusion were used to screen for the optimal brain ischemia model in rats. Model parameters of 300 g weight, 3040 thread bolt, and 3 h brain infarct time were the most suitable for the largest cerebral infarction, highest Longa&#39;s score a...

Watch this Scientific Journal Video about A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats at JoVE.com

A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats

The&#160;purpose of this study is to establish and validate an animal model for research in the recovery and sequela stages of brain ischemia by testing brain infarction and sensorimotor function after middle cerebral artery occlusion/reperfusion (MCAO/R) after 1-90 days in rats.

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery ...

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