eoe sub articles

EoE Sub Articles

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Administration of a repetitive traumatic brain injury (TBI) induction
NOTE: The TBI parameters are adjusted from previous reports, in which the TBI impact was delivered once. The protocol here applies the same parameter, except increasing the total impact number to 10.
<ol>
	<li>TBI equipment
	<ol>
		<li>Use a custom-built portable device to administer a closed-skull TBI (Figure 1A) to the mouse head on the right side, as indicated in Figure 1B.</li>
	</ol>
	</li>
	<li>Preparation before surgery
	<ol>
		<li>Autoclave surgery equipment, including regular scissors, a surgical caliper, surgical spring scissors, surgical forceps, mosquito forceps, headpost pieces, gauze, and cotton-tipped applicators.</li>
		<li>Weigh and record the mouse&#39;s body weight 2 h before surgery. To minimize brain edema during cranial window implantation, inject dexamethasone sodium phosphate at a dose of 4.8 mg/kg [2 mg/mL, usually ~70 &#181;l (need to inject at two sites)] into the quadriceps muscle using an insulin syringe. After the injection of dexamethasone, but prior to starting TBI surgery, prepare the glass windows as indicated in step 2.1 for later use.</li>
		<li>Disinfect the surgical stage and TBI equipment using 75% ethanol. Expand the disposable sterile surgical drape to cover the surgery stage on the stereotaxic platform. Turn on the heating apparatus and the temperature monitor, setting the target temperature to 37 &#176;C.</li>
		<li>Open the oxygen tank valve and adjust the oxygen flow to 1.5 L/min. Open the anesthesia machine and set the isoflurane level value to three. 
		&#8203;NOTE: 100% pure oxygen can help the animal to survive the TBI impacts.</li>
		<li>Place the mouse in the induction chamber and let it stay for 5 minutes to achieve full anesthesia.</li>
		<li>Once the mouse is fully anesthetized (i.e., slow but steady breathing rates at approximately one cycle per 2 s, coupled with the absence of a tail-pinch reflex), remove the mouse from the induction chamber and place the mouse head in the stereotaxic frame.</li>
		<li>Place the anesthesia tubing nose cone over the mouse&#39;s snout.</li>
		<li>Maintain anesthesia using 1.5% isoflurane mixed with 100% pure oxygen at a 1 L/min flow rate until the end of surgery. Periodically check the breath frequency and deliver a tail or toe pinch at least every 15 min throughout surgery to assess appropriate anesthesia. Adjust isoflurane concentration as appropriate to maintain the desired depth of anesthesia.</li>
	</ol>
	</li>
	<li>TBI surgery
	<ol>
		<li>Administer buprenorphine (1 mg/kg, subcutaneously) using disposable insulin syringes and apply ophthalmic ointment to the mouse&#39;s eyes.</li>
		<li>Remove the hair from the top of the head by trimming it with scissors 1 and then applying a depilatory agent for 1 min. 
		&#8203;CAUTION: Do not apply the depilatory agent product for more than 3 minutes to the scalp, as it is a skin irritant. Also, avoid getting any depilatory agent into the mouse&#39;s eyes.</li>
		<li>Clean the scalp skin by applying gauze and cotton-tipped applicators. Then, disinfect the skin, using 75% ethanol first and then betadine. Repeat this three times (i.e., ethanol followed by betadine), leaving the final application of betadine on the skin.</li>
		<li>Re-check the anesthetic depth to ensure that the mouse is fully anesthetized (i.e., slow but steady breathing rates at approximately one cycle per 2 s, coupled with the absence of a tail-pinch reflex). Tent the skin with surgical forceps 3 and make a 12-15 mm long midline incision starting approximately 3 mm posterior from the eyes.</li>
		<li>Excise the skin over the left and right hemisphere of the skull using spring scissors 2.</li>
		<li>Once the skull is exposed, remove the periosteum by gently rubbing it with a sterile cotton-tipped applicator and flushing it with sterile saline.</li>
		<li>Visually inspect the condition of the skull to ensure that it is intact, except for the two small holes, which were made for the previous virus injection surgery.</li>
		<li>Dry the skull area and mark the TBI impact site at the following coordinates: 2.5 mm posterior to the Bregma and 2 mm lateral from the sagittal suture to the right (Figure 1B).</li>
		<li>Quickly remove the mouse from the stereotaxic frame and place the head on the buffer cushion under the TBI device.</li>
		<li>Align the impactor tip with the marked impact site.</li>
		<li>Lift the metal column by pulling the tethered nylon string 15 cm above the mouse head and then release it, allowing the weight to fall freely onto the transducer rod, which is in contact with the skull top at the TBI site. Do not touch the mouse head when delivering the TBI impact.</li>
		<li>Move the mouse onto the heating blanket and place it on its back while monitoring the breathing status.</li>
		<li>Once the mouse rights itself from a supine to a prone position, place it in the isoflurane induction chamber for ~5 min with 3% isoflurane mixed with 100% pure oxygen at a 1.5 L/min flow rate.</li>
		<li>Once the mouse is fully sedated (i.e., slow but steady breathing rates at approximately one cycle per 2 s, coupled with the absence of a tail-pinch reflex), repeat steps 1.3.9-1.3.14 to achieve a total of 10 impacts. 
		&#8203;NOTE: Ten TBI impacts have been found to induce robust phenotype with low mouse mortality in our study (data not published yet). The parameters may be adjusted to achieve a different severity of brain injury by increasing or decreasing the number of impacts and/or the height from which the weight is released. All parameters are subject to approval by local IACUC.</li>
		<li>Check the skull under a surgical microscope and remove the mouse from the study if a skull fracture has occurred.</li>
		<li>For sham surgery, follow the same procedures as described above, including placing the animal under the impactor but without delivering the TBI impacts.</li>
		<li>After the mouse rights itself from a supine to a prone position following the 10th TBI impact, place it in the isoflurane induction chamber for ~5 min. Then, follow steps 1.2.6-1.2.8 to place the mouse head onto the stereotaxic frame for the cranial window implantation surgery described below.</li>
	</ol>
	</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adjustable Precision Applicator Brushes</td>
			<td>Parkell</td>
			<td>S379</td>
			<td></td>
		</tr>
		<tr>
			<td>BD insulin syringe</td>
			<td>BD</td>
			<td>NDC/HRI#08290-3284-38</td>
			<td>5/16&#34; x 31G</td>
		</tr>
		<tr>
			<td>Betadine</td>
			<td>Purdue</td>
			<td>NDC67618-151-17</td>
			<td>Including 7.5% povidone iodine</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>PAR Pharmaceutical</td>
			<td>NDC 42023-179-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethonal</td>
			<td>Fisher Scientific</td>
			<td>04355223EA</td>
			<td>75%</td>
		</tr>
		<tr>
			<td>Heating apparatus</td>
			<td>CWE</td>
			<td>TC-1000 Mouse</td>
			<td>Equiped with the stereotaxic instrument and be used while operating surgery</td>
		</tr>
		<tr>
			<td>Heating blanket</td>
			<td>CVS pharmacy</td>
			<td>E12107</td>
			<td>Extra heating device and be used after surgery</td>
		</tr>
		<tr>
			<td>Lubricant ophthalmic ointment</td>
			<td>Picetal</td>
			<td>NDC 46066-753-55</td>
			<td></td>
		</tr>
		<tr>
			<td>Marker pen</td>
			<td>Delasco</td>
			<td>SMP-BK</td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito forceps</td>
			<td>CAROLINA</td>
			<td>Item #:625314</td>
			<td>Stainless Steel, Curved, 5 in</td>
		</tr>
		<tr>
			<td>Depilatory agent</td>
			<td>McKesson Corporation</td>
			<td>N/A</td>
			<td>Nair Hair Aloe &#38; Lanolin Hair Removal Lotion</td>
		</tr>
		<tr>
			<td>Regular scissor 1</td>
			<td>Eurostat</td>
			<td>eurostat es5-300</td>
			<td></td>
		</tr>
		<tr>
			<td>Regular scissor 2</td>
			<td>World Precision Instruments</td>
			<td>No. 501759-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Bioworld</td>
			<td>L19102411PR</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring scissor 1</td>
			<td>World Precision Instruments</td>
			<td>No. 91500-09</td>
			<td>Tip straight</td>
		</tr>
		<tr>
			<td>Spring scissor 2</td>
			<td>World Precision Instruments</td>
			<td>No. 91501-09</td>
			<td>Tip straight</td>
		</tr>
		<tr>
			<td>Stereotaxic platform</td>
			<td>KOPF</td>
			<td>Model 900LS</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical forceps 1</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>Catlog# 0508-5/45-PO</td>
			<td>Style 5/45, curved</td>
		</tr>
		<tr>
			<td>Surgical forceps 2</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>catlog# 0103-5-PO</td>
			<td>Style 5, straight</td>
		</tr>
		<tr>
			<td>Surgical forceps 3</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>catlog# 72912</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical forceps 4</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>Catlog# 0508-5/45-PO</td>
			<td>Style 5/45, curved</td>
		</tr>
		<tr>
			<td>Surgical lamp</td>
			<td>Leica</td>
			<td>Leica KL300 LED</td>
			<td></td>
		</tr>
		<tr>
			<td>Vetbond Tissue Adhesive</td>
			<td>3M Animal Care</td>
			<td>Part Number:014006</td>
			<td></td>
		</tr>
	</tbody>
</table>

delivery of a traumatic brain injury to a mouse brain

Source: Zhong, J., et al. <a href="https://app.jove.com/v/64701">Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope.</a> J. Vis. Exp. (2023)
This video demonstrates the delivery of Traumatic Brain Injury (TBI) in mice. After exposing the skull, the TBI site is marked. The mouse is then positioned under the TBI device. The impact is delivered by lifting the metal rod and allowing it to drop freely onto the impactor, which is in contact with the skull.

<img alt="Figure 1" src="/files/ftp_upload/22967/22967_Figure1.jpg" /> 
Figure 1: Diagrams for the TBI device, TBI impact site, and window preparation. (A) Equipment for the weight-drop TBI device. 1: pedestal, 2: bracket, 3: adjustable clamp, 4: nylon string, 5: weight dropping tunnel, 6: metal weight column (50 g), 7: impactor, 8: buffer cushion. (B) A schematic of the virus injection and TBI impact site, with the coordinates 2.5 mm posterior to the Bregma and 2 mm...

Delivery of a Traumatic Brain Injury to a Mouse Brain

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

Begin with an anesthetized mouse with its head secured in a stereotaxic apparatus.
Apply ointment to prevent eye dryness.
Remove the hair from the head and disinfect the area.
Make a midline incision on the scalp and remove the connective tissue to expose the skull.
Clean the area with saline and allow the skull to dry.
Using the point where the skull bones join and the bone boundary as a reference, mark the desired area for the traumatic brain injury or TBI.
Position the mouse under the TBI device.
The TBI device consists of an impactor and a metal column within a tunnel.
Align the impactor tip with the marked area on the skull.
Lift the metal column to a desired height and release it freely to deliver an impact on the skull, inducing a TBI at the specified location.

delivery-of-a-traumatic-brain-injury-to-a-mouse-brain

Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Trauma

70 次观看. 来源:Zhong， J. 等人 使用双光子显微镜对闭合颅骨创伤性脑损伤和颅窗小鼠荧光蛋白表达进行活体成像。J. Vis. Exp. （2023 年） 该视频演示了小鼠创伤性脑损伤 （TBI） 的实施。暴露颅骨后，标记 TBI 部位。然后将鼠标放置在 TBI 设备下方。通过提起金属棒并使其自由落到与颅骨接触的撞击器上来产生冲击。 

视频: 将创伤性脑损伤输送到小鼠大脑 - 概念

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the detonation of explosive devices, however, the mechanisms that underlie the brain damage resulting from blast overpressure exposure are not entirely understood and are believed to be unique to this type of brain injury. Preclinical models are crucial tools that contribute to better understand blast-induced brain injury. A novel in vitro blast TBI model was developed using an open-ended shock tube to simulate real-life open-field blast waves modelled by the Friedlander waveform. C57BL/6N mouse organotypic hippocampal slice cultures were exposed to single shock waves and the development of injury was characterized up to 72 h using propidium iodide, a well-established fluorescent marker of cell damage that only penetrates cells with compromised cellular membranes. Propidium iodide fluorescence was significantly higher in the slices exposed to a blast wave when compared with sham slices throughout the duration of the protocol. The brain tissue injury is very reproducible and proportional to the peak overpressure of the shock wave applied.

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

一种新的脑外伤体损伤模型

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

科研

JoVE Journal

神经科学

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

小鼠轻度创伤性脑损伤的电磁控制闭头模型

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

在 mTBI 大鼠模型中验证损伤反应和行为评估

Rat Model of Closed-Head Mild Traumatic Injury and its Validation

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful insights, developing a stable and reproducible animal model is essential. In this study, we report a detailed description of a closed-head mTBI model and a representative validation method using Sprague-Dawley rats to verify the modeling effect. The model involves dropping a 550 g mass weight from a height of 100 cm directly onto the head of a rat on a destructible surface, followed by a 180-degree turn. To assess the injury, rats underwent a series of neurobehavioral assessments 10 min post-injury, including time of loss of consciousness, first seeking-behavior time, escape ability, and beam balance ability test. During the acute and subacute stages following the injury, behavioral tests were conducted to assess motor coordination ability (Beam task), anxiety (Open Field test), and learning and memory abilities (Morris Water Maze test). The closed-head mTBI model produced a consistent injury response with minimal mortality and replicated real-life situations. The validation method effectively verified the model development and ensured the stability and consistency of the model.

作者聚焦:推进创伤性脑损伤研究 - 用于准确复制和快速评估的闭头模型 

Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages.

闭合头轻度创伤大鼠模型及其验证

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful ...

Delivery of a Traumatic Brain Injury to a Mouse Brain

成績單

Delivery of a Traumatic Brain Injury to a Mouse Brain