inducing ischemic stroke via middle cerebral artery occlusion using a thread bolt

Inducing Ischemic Stroke via Middle Cerebral Artery Occlusion Using a Thread Bolt

Begin by placing an anesthetized rat on a surgical fixing table. Throughout the surgery, maintain the body temperature of the rats at 37 degrees Celsius in a small animal thermostat. Connect the mouth to the anesthesia machine mask and confirm deep anesthesia by lack of extremity tension, corneal reflexes, and pain.
Fix the rat&#39;s limbs to the operating table using paper bandages. With an electric shaver, remove the neck coat and sterilize the neck with 75% alcohol. Use ophthalmic scissors to cut 2 to 3 centimeters along the central longitudinal shape of the neck. And then, with ophthalmic forceps, separate the subcutaneous muscle. Use homemade retractor to fully expose the field of vision.
Care must be taken to present appropriate aseptic technique for all survival surgical procedures. The technique illustrated later in the video should be practiced through the entire procedure. Use the micro-forceps to isolate the common carotid artery, the external carotid artery, and the internal carotid artery.
Working under the microscope, use 8 0 sutures to ligate the common carotid artery with hard knot, external carotid artery far from the heart end with hard knot, internal carotid artery with loose knot, and then to line the external carotid artery near the heart end. Using microscissors, cut a small opening in the external carotid artery and gently insert a thread bolt.
Ligate the suture of the external carotid artery that has been in loose knot and cut off the external carotid artery. Loosen the loose knot of the internal carotid artery, and continue inserting the thread bolt to the beginning of the middle cerebral artery, marked with the suture, and cut off the exposed thread bolt.

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Establishment of a unilateral middle cerebral artery occlusion/reperfusion (MCAO/R) model in rats
NOTE: During the operation, use the microforceps gently to prevent breakage of the blood vessel. Avoid damage to the nerves and other blood vessels in the neck of the rat when the vessel is isolated. Care must be taken to present appropriate aseptic technique for all survival surgical procedures. The technique illustrated later in the video should be practiced through the entire procedure.
<ol>
	<li>Throughout the surgery, maintain the body temperature of the rats at 37.0 &#177; 0.5 &#176;C in a small animal thermostat. Prepare four 6 cm 5-0 sutures.</li>
	<li>Set the oxygen flow rate of a small animal anesthesia machine (with a waste gas treatment device) at 0.4&#8722;0.6 L/min and the concentration of isoflurane to be 5%. Place the rat in the anesthesia machine.</li>
	<li>After the animal has fainted, place the rat on a surgical fixing table. Connect the mouth of the rat to the mask of the anesthesia machine (the oxygen flow rate remains unchanged; adjust the concentration of isoflurane to be 3%). Confirm that the animal has entered deep anesthesia by observing a lack of extremity tension, corneal reflexes, and pain.</li>
	<li>Fix its limbs to lie on the operating table with paper bandages (or other tools).</li>
	<li>Remove the neck coat with an electric shaver and sterilize with 75% alcohol (iodophor is better). Fix the mouth of the rat with a hook.</li>
	<li>Cut 2&#8722;3 cm along the central longitudinal shape of the neck with ophthalmic scissors.</li>
	<li>Separate the common carotid artery. Separate the subcutaneous muscle with ophthalmic forceps. Use a homemade retractor to fully expose the field of vision. After separating the anterior muscle of the trachea with ophthalmic forceps, separate along the right sternocleidomastoid tendon until the common carotid artery is visible.</li>
	<li>Isolate the common carotid artery, the external carotid artery, and the internal carotid artery with ophthalmic forceps. Ligate the common carotid artery (hard knot), external carotid artery far from the heart end (hard knot), and internal carotid artery (loose knot) with 5-0 sutures. Line the external carotid artery near the heart end with 5-0 sutures.</li>
	<li>Insert a thread bolt. Cut a small opening in the external carotid artery using ophthalmic scissors and gently insert a thread bolt. Ligate the suture of the external carotid artery that has been in a loose knot and cut off the external carotid artery.
	<ol>
		<li>Loosen the loose knot of the internal carotid artery and continue inserting the thread bolt to the beginning of the middle cerebral artery (suture marked). Then, cut off the exposed thread bolt.</li>
	</ol>
	</li>
	<li>After the ischemic time is reached (2-3 h), fix the fracture of the external carotid artery with one microforceps and gently pull out the thread bolt with another microforceps. When the front end of the thread bolt is completely withdrawn from the internal carotid artery, ligate the external carotid artery that was lined with 5-0 sutures and then pull out the thread bolt completely.</li>
	<li>Loosen the common carotid artery, and daub ~50,000 U of penicillin sodium powder on the surface of the wound to prevent infection. Suture subcutaneous muscles and skin with 4 sutures.</li>
	<li>Give ~0.2 mL of sterile saline to the rat orally using a 1 mL syringe (SQ-PEN injection is better) to prevent postoperative water shortage after placing the rat back in the cage.</li>
</ol>

<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>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>Image Pro Plus Software</td>
			<td>Media Cybernetics Inc.</td>
			<td>6</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>Ophthalmic 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>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>
	</tbody>
</table>

Source: Liu, P., et al., <a href="https://app.jove.com/v/60166">A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats.</a> J. Vis. Exp. (2019)
This video demonstrates the process of inducing ischemic stroke in a rat model by occluding the middle cerebral artery (MCA) using a thread bolt. The procedure involves ligating the carotid arteries, inserting a thread bolt into the external carotid artery, and blocking blood flow from the MCA to the brain, leading to neuronal death and mimicking an ischemic stroke.

Source: Liu, P., et al., A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats. J. Vis. Exp. (2019)
This video demonstrates the process ...

Start with an anesthetized, immobilized rat with the dissected neck area exposing the arteries.
Locate the right common carotid artery or CCA, which bifurcates into the external carotid artery or ECA and the internal carotid artery or ICA, with branches into the middle cerebral artery or MCA.
Ligate the CCA tightly to reduce blood flow.
Next, tie a tight knot on the distal ECA to prevent blood backflow and a loose knot on the ICA.
Place another loose suture around the proximal ECA.
Make a small incision between the ECA knots and insert a thread bolt, a thin filament with a silicone tip.
Then, tighten the loose knot on the ECA to secure the thread bolt and cut the ECA.
Now, loosen the ICA knot and advance the thread bolt until it blocks the MCA.
This blockage restricts blood flow to the brain, inducing neuronal death and resulting in ischemic stroke.

14 visualizações. Inducing Ischemic Stroke via Middle Cerebral Artery Occlusion Using a Thread Bolt

Vídeo: Inducing Ischemic Stroke via Middle Cerebral Artery Occlusion Using a Thread Bolt - Experimento

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema otimizado para Monitorização de perfusão cerebral no modelo do rato do curso de oclusão da artéria cerebral média intraluminal

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

JoVE Journal

Medicina

A Middle Cerebral Artery Occlusion Technique for Inducing Post-stroke Depression in Rats

Post-stroke depression (PSD) is the most recurrent of all psychiatric complications resulting from an ischemic stroke. A greater majority (about 60%) of all ischemic stroke patients suffer from PSD, a disorder considered to be an ischemic stroke-related precursor for increased death and degradation in health. The pathophysiology of PSD is still obscure. To study the mechanism of development and occurrence of PSD further, and to find out a therapy, we attempted to develop a new protocol that requires occluding the middle cerebral artery (MCA) via the internal carotid artery (ICA) in rats. This protocol describes a model of PSD induced in rats through the middle cerebral artery occlusion (MCAO). Also used in the experiment are the Porsolt forced swim test and the sucrose preference test to confirm and evaluate the depressive mood of the rats under investigation. Rather than inserting the catheter through the external carotid artery (ECA), as stipulated for the original procedure, this MCAO technique has the monofilament passing directly through the ICA. This MCAO technique was developed a few years ago and leads to a reduction in mortality and variability. It is generally accepted that the criteria used are preferred in the selection of biological models. The data obtained with this protocol show that this model of MCAO could be a way of inducing PSD in rats and could potentially lead to the understanding of the pathophysiology and the future development of new drugs and other neuroprotective agents.

Here we present a protocol to induce post-stroke depression in rats by occluding the middle cerebral artery via the internal carotid artery. We use the Porsolt forced swim test and the sucrose preference test to confirm and evaluate induced depressive moods.

Uma técnica de oclusão da artéria cerebral média para induzir A depressão pós-AVC em ratos

Post-stroke depression (PSD) is the most recurrent of all psychiatric complications resulting from an ischemic stroke. A greater majority (about 60%) of ...

Comportamento

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

Modelagem de AVC em Camundongos: Oclusão da Artéria Cerebral Média Transitória através da Artéria Carótida Externa

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

Inducing Ischemic Stroke via Middle Cerebral Artery Occlusion Using a Thread Bolt

Transcrição

Inducing Ischemic Stroke via Middle Cerebral Artery Occlusion Using a Thread Bolt