generating a mouse model of traumatic brain injury using a lateral fluid percussion device

Generating a Mouse Model of Traumatic Brain Injury Using a Lateral Fluid Percussion Device

For traumatic brain injury induction, confirm a lack of response to toe pinch in an anesthetized mouse, and apply ointment to the animal&#39;s eyes. After shaving, disinfect the exposed scalp skin with 70% ethanol before incising the scalp in a sagittal plane. Using forceps, retract the incision on both sides and separate the periosteum slightly.
Use a marker to draw a 3 millimeter diameter circle on the right parietal area of the skull, 2 millimeter away from the midline. Use an electric drill to carefully penetrate the skull without damaging the dura. Remove the bone flap to expose a 3 millimeter bone window, and place a plastic injury cannula over the craniotomy.
After cementing the cannula to the skull with dental acrylic, use a 5 milliliters syringe to fill the cannula with sterile 0.9% normal saline, taking care to avoid bubbles. Turn on the oscilloscope and amplifier. Confirm that the high pressure tube of the lateral fluid percussion entry device is free of air bubbles, and deliver about 10 test pulses until the device gives a steady signal.
Adjust the angle of the pendulum starting position to reach a pulse intensity of about two standard atmosphere, and connect the entry cannula to the lateral fluid percussion injury device. Pull the trigger to release the pendulum, inducing the brain injury. Then obtain and transmit a pulse to the dura through the entire closed fluid filled tubing system.
After brain injury induction, remove the cannula and suture the incision. Then, place the mouse on a heating pad with monitoring until it is ambulatory, before returning the animal to its home cage with ad libitum access to food and water.

generating-mouse-model-traumatic-brain-injury-using-lateral-fluid

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JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Trauma

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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. Animal care
<ol>
	<li>Use 5- to 6-week-old male C57BL/6J mice (20-25 g of weight) in this experiment.</li>
	<li>Maintain mice on a 12 h/12 h light/dark cycle, and make sure they receive food and water ad libitum. Provide the same amounts of food and water to both the sham and TBI groups throughout the study.</li>
	<li>Make all efforts to minimize the animal pain and discomfort.</li>
</ol>
2. Induction of traumatic brain injury
<ol>
	<li>Inject Ketamine (80-100 mg/kg)/Xylazine (10 mg/kg) IP for anesthesia. Test the depth of anesthesia using an eye reflex or pain reflex. Use artificial tears or lubricant eye ointment to keep the eyes from drying.</li>
	<li>After anesthesia, put the mouse in a prone position. Use a temperature-controlled heating pad to maintain the temperature at 37 &#176;C during the surgery and for 30 min after TBI.</li>
	<li>Shave the hair of the incision area.</li>
	<li>Disinfect the scalp with 70% ethanol using three alternating scrubs, then incise the scalp in a sagittal plane.</li>
	<li>Use forceps to retract the incision on both sides and separate the periosteum slightly.</li>
	<li>Use a marker to draw a circle (3 mm diameter) on the right parietal area of the skull, 2 mm away from the midline.</li>
	<li>Drill the skull with an electric drill. Ensure that this step is operated carefully to protect the dura from being damaged.</li>
	<li>Remove the bone flap and expose a small bone window (3 mm in diameter).</li>
	<li>Place a plastic injury cannula (internal diameter = 2.5 mm, length = 8 mm) over the craniotomy and cement the cannula to the skull using a dental acrylic.</li>
	<li>Fill the cannula with sterile 0.9% NaCl (normal saline) using a syringe (5 mL) to ensure that there are no bubbles in the cannula.</li>
	<li>Turn on the oscilloscope and amplifier and ensure that the high-pressure tube of the LFPI device is free of air bubbles. Test the device by delivering about 10 pulses until it gives a steady signal. Adjust the angle of the pendulum starting position to reach a pulse intensity of about 2.0 atm.</li>
	<li>Connect the injury cannula to the LFPI device. Induce brain injury by pulling the trigger and releasing the pendulum. Then, obtain a pulse and transmit it to the dura through the entire closed fluid-filled tubing system.</li>
	<li>Operate the mice in the sham group with the same surgical procedure. Do not perform the LFPI.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Heating pad</td>
			<td>Shanghai SAFE Biotech Co.</td>
			<td>TR-200</td>
			<td>Heating pad</td>
		</tr>
		<tr>
			<td>Injector</td>
			<td>The First Affiliated Hospital, School of Medicine, Zhejiang University</td>
			<td></td>
			<td>Injector</td>
		</tr>
		<tr>
			<td>LFPI device</td>
			<td>Virginia 
			Commonwealth University</td>
			<td>FP302</td>
			<td>LFPI device</td>
		</tr>
		<tr>
			<td>Micro cranial drill</td>
			<td>RWD Life Science</td>
			<td>78061</td>
			<td>Micro cranial drill</td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>The First Affiliated Hospital, School of Medicine, Zhejiang University</td>
			<td></td>
			<td>Povidone Iodine</td>
		</tr>
	</tbody>
</table>

Source: Wen, L., et al. <a href="https://app.jove.com/v/59410">Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice.</a> J. Vis. Exp. (2019).
The video demonstrates a method to generate a mouse model of traumatic brain injury (TBI) using a lateral fluid percussion device (LFPI). Upon anesthetizing the mouse, the brain is exposed by creating a window on the skull. A saline-filled cannula is attached to the window and is then connected to the LFPI. The pendulum of the device is released to create a pressure pulse that inflicts a traumatic brain injury.

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animal ...

Take an anesthetized mouse and apply eye ointment to prevent its eyes from drying.
Shave the head, disinfect the scalp, and incise in the sagittal plane to expose the skull.
Remove the connective tissue layer and mark the target area for injury.
Drill to create a bone flap, then remove it to expose the dura mater, the brain&#39;s outer covering.
Place a cannula over the dura mater and secure it with dental cement.
Fill the cannula with saline, ensuring no air bubbles form, and connect it to the tubing of the lateral fluid percussion device, containing a pendulum and a fluid-filled reservoir.
Release the pendulum from a calibrated angle to strike the reservoir, generating a pressure pulse that travels through the fluid and reaches the brain.
The mechanical stress damages neurons, causing traumatic brain injury.
Remove the cannula, suture the incision, and allow the mouse to recover for further analysis.

13 次观看. 使用侧向液体叩击装置生成创伤性脑损伤的小鼠模型

视频: 使用侧向液体叩击装置生成创伤性脑损伤的小鼠模型 - 实验

Controlled Cortical Impact Model of Mouse Brain Injury with Therapeutic Transplantation of Human Induced Pluripotent Stem Cell-Derived Neural Cells

Traumatic brain injury (TBI) is a leading cause of morbidity and mortality worldwide. Disease pathology due to TBI progresses from the primary mechanical insult to secondary injury processes, including apoptosis and inflammation. Animal modeling has been valuable in the search to unravel injury mechanisms and evaluate potential neuroprotective therapies. This protocol describes the controlled cortical impact (CCI) model of focal, open-head TBI. Specifically, parameters for producing a mild unilateral cortical injury are described. Behavioral consequences of CCI are analyzed using the adhesive tape removal test of bilateral sensorimotor integration. Regarding experimental therapy for TBI pathology, this protocol also illustrates a process for transplanting cultured cells into the brain. Neural cell cultures derived from human induced pluripotent stem cells (hiPSCs) were chosen for their potential to show superior functional restoration in human TBI patients. Chronic survival of hiPSCs in the host mouse brain tissue is detected using a modified DAB immunohistochemical process.

This protocol demonstrates methodologies for a mouse model of open-skull traumatic brain injury and transplantation of cultured human induced pluripotent stem cell-derived cells into the injury site. Behavioral and histologic tests of outcomes from these procedures are also described in brief.

人类诱导多能干细胞衍生神经细胞治疗移植小鼠脑损伤控制皮质影响模型

Traumatic brain injury (TBI) is a leading cause of morbidity and mortality worldwide. Disease pathology due to TBI progresses from the primary mechanical ...

科研

JoVE Journal

发育生物学

Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury

Competition over resources such as food, territory, and mates significantly influences relationships within animal species and is mediated through social hierarchies that are often based on dominant-submissive relationships. The dominant-submissive relationship is a normal behavioral pattern among the individuals of a species. Traumatic brain injury is a frequent cause of social interaction impairment and the reorganization of dominant-submissive relationships in animal pairs. This protocol describes submissive behavior in adult male Sprague-Dawley rats after the induction of traumatic brain injury using a fluid-percussion model compared to naive rats through a series of dominant-submissive tests performed between 29 days and 33 days after induction. The dominant-submissive behavior test shows how brain injury can induce submissive behavior in animals competing for food. After traumatic brain injury, the rodents were more submissive, as indicated by them spending less time at the feeder and being less likely to arrive first at the trough compared to the control animals. According to this protocol, submissive behavior develops after traumatic brain injury in adult male rats.

The present protocol describes a rat model of fluid percussion-induced traumatic brain injury followed by a series of behavioral tests to understand the development of dominant and submissive behavior. Using this model of traumatic brain injury in conjunction with specific behavioral tests enables the study of social impairments following brain injury.

评估创伤性脑损伤后成年大鼠的显性顺从行为

Competition over resources such as food, territory, and mates significantly influences relationships within animal species and is mediated through social ...

行为学

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

创伤性脑损伤诱导受控皮质冲击的母模型

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

Generating a Mouse Model of Traumatic Brain Injury Using a Lateral Fluid Percussion Device

成績單

Generating a Mouse Model of Traumatic Brain Injury Using a Lateral Fluid Percussion Device