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. Arterial catheterization
<ol>
	<li>Anesthetize the mouse with an intraperitoneal injection of ketamine and xylazine and an analgesic according to local practice (e.g., buprenorphine). Pinch the hind limbs with forceps to confirm the correct depth of anesthesia. 
	NOTE: For this study, male C57BL/6N mice at the age of 8&#8722;12 weeks were used (weight 20&#8722;25 g). The authors have tested male BALB/c of the same age unsuccessfully but have not tried other strains.</li>
	<li>Remove the hair in the regions of interest (head, neck) using an electric razor. Ensure that no loose hair remains to prevent wound contamination. Subsequently, disinfect the surgical field with 70% ethanol/chlorhexidine/ betadine and place the mouse supine with the head facing the surgeon.</li>
	<li>Make a cervical midline incision with dissecting scissors.</li>
	<li>Dissect the submandibular glands and neck muscle tissue and separate them in order to expose the common carotid artery. Use mostly blunt dissection via forceps.</li>
	<li>Place three 8-0 silk ligatures beneath the right common carotid artery.</li>
	<li>Place a clamp on the proximal ligature and bring tension into the artery so that the flow is suspended.</li>
	<li>Close the most distal ligature.</li>
	<li>Insert the arterial catheter through a small, preformed skin hole on the cranial aspect of the incision. Squeeze and deform the lumen of the catheter if it appears too large to reduce blood backflow. Fixate with all three ligatures.</li>
	<li>Fixate the catheter to the skin to avoid dislocation. Do this by using a suture (e.g., 5-0 monofilament, non-absorbable) that connects the catheter to the skin in the area of the preformed skin hole.</li>
</ol>
2. Tracheostomy
<ol>
	<li>Dissect the pre-tracheal musculature bluntly using forceps.</li>
	<li>Place two 8-0 silk ligatures beneath the trachea.</li>
	<li>Tracheotomize using micro scissors as proximally as possible to avoid unilateral ventilation. Use a horizontal cutting line between two tracheal cartilages.</li>
	<li>Insert the ventilation tube and fixate with both prepared ligatures. 
	NOTE: The proximal end of the trachea does not need to be ligated.</li>
	<li>Close the skin with a running suture (e.g., 6-0 monofilament, non-absorbable).</li>
	<li>Ventilate the mouse with a frequency of 150/min and a tidal volume of 200 &#181;L.</li>
</ol>
3. Brain death (BD) induction
<ol>
	<li>Arrange the mouse to the prone position.</li>
	<li>Remove the skin from the skull using surgical scissors and forceps to hold the skin.</li>
	<li>Drill a 1 mm caliber borehole paramedially above the left parietal cortex. Stop drilling before breaching the inner compact bone and the dura mater.</li>
	<li>Penetrate the final tissue bridge of the skull using blunt forceps, removing sharp edges.</li>
	<li>Insert the balloon catheter, so that it is entirely within the cranial cavity. Ensure that the balloon is prefilled with saline and all air is evacuated.</li>
	<li>Begin inflation at ~0.1 mL/min over a period of 10&#8722;15 min (total volume of 0.8&#8722;1.2 mL) with the help of a syringe pump. 
	NOTE: The mouse will exhibit myoclonus, mydriasis, further seizure activity, and agonal gasps.</li>
	<li>Pronounce the mouse brain dead once the tail of the mouse has gone stiff and erected. 
	NOTE: BD is confirmed by a characteristic initial blood pressure peak (Cushing reflex), the absence of brain stem reflexes, and spontaneous breathing. Regular apnea testing should be avoided during experiments, as mice may become circulatory unstable due to a lack of oxygen.</li>
	<li>Stop the inflation of the balloon catheter.</li>
	<li>Put a heating blanket over the mouse to avoid hypothermia.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Arterial catheter (BD Neoflon 26G)</td>
			<td>BD</td>
			<td>391349</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood Pressure Transducers (APT300)</td>
			<td>Harvard Apparatus Inc.</td>
			<td>73-3862</td>
			<td></td>
		</tr>
		<tr>
			<td>Fogarty Arterial Embolectomy Catheter N&#176; 3</td>
			<td>Edwards Lifesciences Corporation</td>
			<td>120403F</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>FST</td>
			<td>11271-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Homeothermic Blanket Systems with Flexible Probe</td>
			<td>Harvard Apparatus Inc.</td>
			<td>55-7020</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketansol</td>
			<td>Graeub</td>
			<td>6680110</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissor</td>
			<td>FST</td>
			<td>15018-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FST</td>
			<td>12060-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene 5-0</td>
			<td>Ethicon</td>
			<td>8698H</td>
			<td></td>
		</tr>
		<tr>
			<td>Pump 11 Elite Infusion Only Single</td>
			<td>Harvard Apparatus Inc.</td>
			<td>70-4500</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissor</td>
			<td>FST</td>
			<td>14075-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotactic microscope</td>
			<td>Olympus</td>
			<td>SZX7</td>
			<td></td>
		</tr>
		<tr>
			<td>Transpore Tape</td>
			<td>3M</td>
			<td>1527-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Underpads</td>
			<td>Molinea.A</td>
			<td>274301</td>
			<td></td>
		</tr>
		<tr>
			<td>Ventilator for mice (MiniVent Model 845)</td>
			<td>Harvard Apparatus Inc.</td>
			<td>73-0043</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylasol</td>
			<td>Graeub</td>
			<td>7630109</td>
			<td></td>
		</tr>
	</tbody>
</table>

inducing brain death in a mouse model using a balloon catheter

Source: Ritschl, P. V., et al. &#160;<a href="https://appjove.unicartagenaproxy.elogim.com/v/60831">Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy.</a> J. Vis. Exp. (2020).
This video demonstrates the procedure for inducing brain death in a mouse model by increasing controlled intracranial pressure through a balloon catheter, mimicking natural brain death.

Inducing Brain Death in a Mouse Model Using a Balloon Catheter

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 in the prone position, with its head shaved.
Make an incision in the skin over the head.
Drill a small hole in the skull, stopping just before the brain layer or dura mater to avoid damaging brain tissue.
Use blunt forceps to penetrate the final tissue bridge and remove any sharp edges.
Insert a balloon catheter through this opening, ensuring it is fully within the cranial cavity. This catheter is prefilled with saline and free of air.
Infuse additional saline through the catheter to gradually inflate the balloon, increasing intracranial pressure.
As the pressure rises, it compresses the brain&#39;s blood vessels, restricting blood flow and oxygen to neurons, leading to neuronal death.
This causes the mouse&#39;s tail to stiffen and breathing to stop, indicating brain death.

inducing-brain-death-in-a-mouse-model-using-a-balloon-catheter

Universidad de Cartagena UDEC

Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Trauma

34 Views. Fonte: Ritschl, P. V., et al. Induzione della morte cerebrale nei topi utilizzando il monitoraggio della pressione sanguigna intra-arteriosa e la ventilazione tramite tracheostomia. J. Vis. Exp. (2020). Questo video dimostra la procedura per indurre la morte cerebrale in un modello murino aumentando la pressione intracranica controllata attraverso un catetere a palloncino, imitando la morte cerebrale naturale. 

Video: Induzione della morte cerebrale in un modello murino utilizzando un catetere a palloncino - Concept

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also demonstrate that traumatic brain injuries cause changes to the gut microbiota. However, mechanisms underlying the bidirectional regulation of the brain-gut axis remain unknown. Currently, few models exist for studying the changes in gut microbiota after traumatic brain injury. Therefore, the presented study combines protocols for inducing traumatic brain injury using a lateral fluid percussion device and analysis of caecum samples following injury for investigating alterations in the gut microbiome. Alterations of the gut microbiota composition after traumatic brain injury are determined using 16S-rDNA sequencing. This protocol provides an effective method for studying the relationships between enteric microorganisms and traumatic brain injury.

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

Indagare sulle alterazioni nel caecum Microbiota dopo una lesione cerebrale traumatica nei topi

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

Ricerca

JoVE Journal

Neuroscienze

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.

Modello Murine di impatto corticale controllato per l'induzione di lesioni cerebrali traumatiche

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

Study of Experimental Organ Donation Models for Lung Transplantation

Experimental models are important tools for understanding the etiological phenomena involved in various pathophysiological events. In this context, different animal models are used to study the elements triggering the pathophysiology of primary graft dysfunction after transplantation to evaluate potential treatments. Currently, we can divide experimental donation models into two large groups: donation after brain death and donation after circulatory arrest. In addition, the deleterious effects associated with hemorrhagic shock should be considered when considering animal models of organ donation. Here, we describe the establishment of three different lung donation models (post-brain death donation, post-circulatory death donation, and post-hemorrhagic shock donation) and compare the inflammatory processes and pathological disorders associated with these events. The objective is to provide the scientific community with reliable animal models of lung donation for studying the associated pathological mechanisms and searching for new therapeutic targets to optimize the number of viable grafts for transplantation.

The present study shows the establishment of three different lung donation models (post-brain death donation, post-circulatory death donation, and post-hemorrhagic shock donation). It compares the inflammatory processes and pathological disorders associated with these events.

Studio di modelli sperimentali di donazione di organi per il trapianto di polmone

Experimental models are important tools for understanding the etiological phenomena involved in various pathophysiological events. In this context, ...

Inducing Brain Death in a Mouse Model Using a Balloon Catheter

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Inducing Brain Death in a Mouse Model Using a Balloon Catheter