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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. Cannulation of the Carotid Artery
The carotid artery cannulation may be performed before or after the cannulation of the mesenteric lymph duct. Some operators prefer to cannulate the carotid artery prior to cannulating the lymph duct so as to reduce potentially dislodging the lymph cannula when moving the animal.
<ol>
	<li>Place a mark on the carotid artery cannula 2.5 cm from the bevel tip to identify the portion of the tubing to insert into the artery. Connect the other end of the cannula (i.e., without bevel) to a 23 or 25-G needle attached to a syringe filled with anticoagulant solution and fill the cannula with the anti-coagulant solution making sure no air gaps are present.</li>
	<li>To facilitate cannulation, place the anesthetized rat in dorsal recumbency with the neck stretched and the head pointing toward the operator. Note that some operators prefer to perform this technique with the rat&#39;s tail pointing toward the operator.</li>
	<li>Place a 1 - 1.5 cm longitudinal incision through the skin layer above the left of the trachea in the sagittal plane.</li>
	<li>Dissect the subcutaneous connective tissue using blunt tip forceps to expose the underlying paired neck muscles.</li>
	<li>Retract the neck muscles to reveal the left carotid artery (located ~1 cm below the skin surface and pulsing). Clean the overlying connective tissue from the artery by blunt dissection.</li>
	<li>Isolate a ~1 cm section of the artery carefully using blunt tissue forceps, taking care not to damage the adjacent vagal nerve tract. Perform this by opening and closing the blunt tip forceps vertically along either side of the artery to free it from the surrounding tissue and vagus nerve.</li>
	<li>Using fine-tip forceps, thread two silk sutures underneath the carotid artery and place them at each end of the isolated artery section. Tie off the suture closest to the operator and furthest from the rat&#39;s heart and clavicle (Figure 1A).</li>
	<li>Occlude blood flow through the artery by placing a pair of straight fine-tip forceps underneath the artery (Figure 1B).</li>
	<li>Using the fine-tip iris scissors, place a small incision on the top surface of the artery (about 1/3 of the way down from the top of the isolated area). Flush any spilled blood on the surface of the artery with heparinized saline and gently wipe using cotton tips.</li>
	<li>Insert the tip of the cannula into the artery with a pair of curved tip forceps. Once inserted, remove the straight fine tip forceps occluding the blood flow and advance the cannula 2.5 cm into the artery using two pairs of forceps, one to hold the cannula inside the artery to stop blood from leaking back and the other to insert the cannula.</li>
	<li>Use a small artery clamp to hold the cannula inside the artery and remove the straight fine tip forceps that were placed below the artery in step 1.8 to occlude the blood flow.</li>
	<li>Confirm the patency of the cannula by injecting an anti-coagulant solution into the cannula and drawing back a small amount of blood (Figure 1C). Then flush the cannula with an anticoagulant solution and seal the end using the flame of a lighter or a cannula plug.</li>
	<li>Tie the cannula in place with the two sutures placed under the artery in step 1.8.</li>
	<li>Remove the artery clamp and secure a third suture around the artery and cannula. Close the neck wound with sutures.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Sterile saline</td>
			<td>Baxter healthcare</td>
			<td>AHB 1307</td>
			<td>Any brand can be used. Example here is Baxter 100 ml saline bags, box of 50</td>
		</tr>
		<tr>
			<td>70 % ethanol in water</td>
			<td>Any</td>
			<td></td>
			<td>Any brand can be used</td>
		</tr>
		<tr>
			<td>Heparin (35000I.U. in 35 mL)</td>
			<td>Sigma Pharmaceuticals</td>
			<td>337220</td>
			<td>http://sigmaco.com.au/</td>
		</tr>
		<tr>
			<td>Polyethylene (PE) cannula o.d. 0.96 mm x i.d. 0.58 mm</td>
			<td>Microtube extensions</td>
			<td>PE8050</td>
			<td>Any brand can be used. Example here is PE tubing 0.8x0.5 mm, 30 m</td>
		</tr>
		<tr>
			<td>Polyethylene (PE) cannula o.d. 0.8 mm x i.d. 0.5 mm</td>
			<td>Microtube extensions</td>
			<td>PE9658</td>
			<td>Any brand can be used. Example here is PE tubing 0.96x0.58 mm, 30 m</td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>Any</td>
			<td></td>
			<td>Any brand can be used</td>
		</tr>
		<tr>
			<td>Markers</td>
			<td>Any</td>
			<td></td>
			<td>Any brand can be used</td>
		</tr>
		<tr>
			<td>Cigarette lighter</td>
			<td>Any</td>
			<td></td>
			<td>Any brand can be used</td>
		</tr>
		<tr>
			<td>23 gauge needles</td>
			<td>Livingstone International</td>
			<td>DN23GX0.75LV</td>
			<td>Any brand can be used. Example here is Livingstone Disposable Needle, Sterile, 23GX0.75inch, 100/BOX. http://www.livingstone.com.au/?PG=search_result&#38;CAT= 
			6&#38;search=DN23GX0.75LV</td>
		</tr>
		<tr>
			<td>25 gauge needles</td>
			<td>Livingstone International</td>
			<td>DN25GX1.0LV</td>
			<td>Any brand can be used. Example here is Livingstone Disposable Needle, Sterile, 25GX1.0inch, 100/BOX. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search= 
			DN25GX1.0LV</td>
		</tr>
		<tr>
			<td>1 ml syringe</td>
			<td>Livingstone International</td>
			<td>T3SS01TA</td>
			<td>Any brand can be used. Example here is Terumo syringe 1 ml Slip Tuberculin 100/Box. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search 
			=T3SS01TA</td>
		</tr>
		<tr>
			<td>10 ml syringe</td>
			<td>Livingstone International</td>
			<td>T3SS10SA</td>
			<td>Any brand can be used. Example here is Terumo syringe 10 ml Slip 100/Box. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search 
			=T3SS10SA</td>
		</tr>
		<tr>
			<td>Gauze swabs</td>
			<td>Livingstone International</td>
			<td>GSC075</td>
			<td>Any brand can be used and cut to required size. Example here is gauze swabs cotton filled 7.5x7.5 cm, 8 ply. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search 
			=GSC075</td>
		</tr>
		<tr>
			<td>Cotton buds</td>
			<td>Livingstone International</td>
			<td>CTAST075DP</td>
			<td>Any brand can be used. Example here is Livingstone cotton applicator plastic double tipped. 75MM. 100/PK. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search 
			=CTAST075DP</td>
		</tr>
		<tr>
			<td>Silk suture</td>
			<td>Livingstone International</td>
			<td>DTSK163019F4</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel blades</td>
			<td>Fine Science Tools (FST)</td>
			<td>10020-00</td>
			<td>Any brand can be used. Example here is FST Scalpel Blade #20. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=191</td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td>Fine Science Tools (FST)</td>
			<td>10004-13</td>
			<td>Any brand can be used. Example here is FST Scalpel Handle #4. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=298&#38;CategoryId=51</td>
		</tr>
		<tr>
			<td>1 x Small surgical scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>14060-09</td>
			<td>Any brand can be used. Example here is FST Fine Scissors, 9 cm with 21 mm cutting edge, sharp, straight. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=40&#38;CategoryId=17</td>
		</tr>
		<tr>
			<td>2 x Forceps with serrated curved tip</td>
			<td>Fine Science Tools (FST)</td>
			<td>11001-13</td>
			<td>Any brand can be used. Example here is FST 13 cm standard pattern forceps with curved 2.8x1.4 mm tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=405&#38;CategoryId=32</td>
		</tr>
		<tr>
			<td>1 x Iridectomy scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>15000-08</td>
			<td>Any brand can be used. Example here is FST Vannas Spring Scissors - 2.5mm Cutting Edge, Straight. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=17&#38;CategoryId=16</td>
		</tr>
		<tr>
			<td>1 x Forceps with straight serated tip</td>
			<td>Fine Science Tools (FST)</td>
			<td>11650-10</td>
			<td>Any brand can be used. Example here is FST Graefe 10 cm straight with serrated 1 x 0.99 mm tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=390&#38;CategoryId=32</td>
		</tr>
		<tr>
			<td>1 x Forceps with smooth sharp straight fine tip</td>
			<td>Fine Science Tools (FST)</td>
			<td>11251-10</td>
			<td>Any brand can be used. Example here is FST Dumont #5 forceps straight 11cm with 0.08 x 0.04mm tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=335&#38;CategoryId=29</td>
		</tr>
		<tr>
			<td>1 x Forceps with smooth fine curved forceps</td>
			<td>Fine Science Tools (FST)</td>
			<td>11063-07</td>
			<td>Any brand can be used. Example here is FST Delicate Forceps 9 cm with smooth 0.4 x 0.3mm tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=360</td>
		</tr>
		<tr>
			<td>1 x Suture needle holder</td>
			<td>Fine Science Tools (FST)</td>
			<td>12001-13</td>
			<td>Any brand can be used. Example here is FST 13cm Hasley Needle Holder with 16 mm length x 1.9 mm width tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=254&#38;CategoryId=70</td>
		</tr>
		<tr>
			<td>1 x Artery clamp</td>
			<td>Fine Science Tools (FST)</td>
			<td>18050-28</td>
			<td>Any brand can be used. Example here is FST Bulldog Serrefines straight, 28 mm long, 9x1.6 mm jaw dimension with medium clamp press. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=270&#38;CategoryId=82</td>
		</tr>
	</tbody>
</table>

cannulation of the carotid artery in a rat model

Source: Trevaskis, N. L. et al., <a href="https://app.jove.com/v/52389">The Mesenteric Lymph Duct Cannulated Rat Model: Application to the Assessment of Intestinal Lymphatic Drug Transport.</a> J. Vis. Exp. (2015)
In this video, we demonstrate a surgical procedure to cannulate the left carotid artery in a rat model.

<img alt="Figure 1" src="/files/ftp_upload/21761/21761_Figure1.jpg" /> 
Figure 1. Photographs of (A) the carotid artery isolated with two silk sutures with the one closest to the operator tied off, (B) the carotid artery isolated and blood flow occluded with a pair of straight fine tip forceps placed underneath the artery, and (C) a carotid artery with the cannula in place and the patency being checked by drawing bac...

Cannulation of the Carotid Artery in a Rat Model

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

Prepare an anesthetized rat in a supine position.
Incise longitudinally through the skin above the anatomical left aspect of the trachea.
Dissect the subcutaneous connective tissue. Retract the neck muscles, revealing the left carotid artery.
Isolate an artery section and place silk sutures around both ends.
Ligate the suture placed distal from the heart to prevent blood flow toward the head region.
Obstruct blood flow through the artery section. Make an incision on the top surface of the artery.
Insert an anticoagulant-filled cannula connected to an anticoagulant-filled syringe into the incised section.
Advancing the cannula to the predetermined depth, inject the anticoagulant solution into the cannula.
Draw back a small blood volume through the cannula to confirm unobstructed flow.
Flush with the anticoagulant solution and seal the cannula&#39;s end to prevent blood leakage.
Secure the cannula within the artery. Close the surgical incision.
The cannulated rat model is ready for experiments.

cannulation-of-the-carotid-artery-in-a-rat-model

Research

JoVE Encyclopedia of Experiments

Immunology

Immunopathology

Assay and Analysis

786 Views. Source: Trevaskis, N. L. et al., The Mesenteric Lymph Duct Cannulated Rat Model: Application to the Assessment of Intestinal Lymphatic Drug Transport. J. Vis. Exp. (2015) In this video, we demonstrate a surgical procedure to cannulate the left carotid artery in a rat model. 

Video: Cannulation of the Carotid Artery in a Rat Model - Concept

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

JoVE Journal

Medicine

A CFSE-based Assay to Study the Migration of Murine Skin Dendritic Cells into Draining Lymph Nodes During Infection with Mycobacterium bovis Bacille Calmette-Gu&#233;rin

Dendritic cells (DCs) are important for initiating immune responses, in part through their ability to acquire and shuttle antigen to the draining lymph node (DLN). The mobilization of DCs to the DLN is complex and remains to be fully elucidated during infection. Herein described is the use of an innovative, simple assay that relies on the fluorochrome 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) to track the migration of DCs during footpad infection with Mycobacterium bovis Bacille Calmette-Gu&#233;rin (BCG) in C57BL/6 mice. This assay enables the characterization of skin DC sub-populations that actively relocate to the draining, popliteal LN in response to BCG. This protocol originates from a BCG model where migratory skin DCs were identified by flow cytometry. The assay is amiable to the study and identification of DCs or other cells that home to the popliteal LN after inoculation of microbes, their metabolites or other inflammatory stimuli in the footpad, and consequently to study factors that regulate the migration of these cells.

A CFSE-based Assay to Study the Migration of Murine Skin Dendritic Cells into Draining Lymph Nodes During Infection with &lt;em&gt;Mycobacterium bovis&lt;/em&gt; Bacille Calmette-Gu&amp;#233;rin

An assay in mice to track cell migration from skin to draining lymph node is described which enables the characterization of skin Dendritic cells mobilized to the lymph node after footpad infection with Bacille Calmette-Gu&#233;rin.

Dendritic cells (DCs) are important for initiating immune responses, in part through their ability to acquire and shuttle antigen to the draining lymph ...

Immunology and Infection

Isolation and Cannulation of Cerebral Parenchymal Arterioles

Intracerebral parenchymal arterioles (PAs), which include parenchymal arterioles, penetrating arterioles and pre-capillary arterioles, are high resistance blood vessels branching out from pial arteries and arterioles and diving into the brain parenchyma. Individual PA perfuse a discrete cylindrical territory of the parenchyma and the neurons contained within. These arterioles are a central player in the regulation of cerebral blood flow both globally (cerebrovascular autoregulation) and locally (functional hyperemia). PAs are part of the neurovascular unit, a structure that matches regional blood flow to metabolic activity within the brain and also includes neurons, interneurons, and astrocytes. Perfusion through PAs is directly linked to the activity of neurons in that particular territory and increases in neuronal metabolism lead to an augmentation in local perfusion caused by dilation of the feed PA. Regulation of PAs differs from that of better-characterized pial arteries. Pressure-induced vasoconstriction is greater in PAs and vasodilatory mechanisms vary. In addition, PAs do not receive extrinsic innervation from perivascular nerves &#8212; innervation is intrinsic and indirect in nature through contact with astrocytic endfeet. Thus, data regarding contractile regulation accumulated by studies using pial arteries does not directly translate to understanding PA function. Further, it remains undetermined how pathological states, such as hypertension and diabetes, affect PA structure and reactivity. This knowledge gap is in part a consequence of the technical difficulties pertaining to PA isolation and cannulation. In this manuscript we present a protocol for isolation and cannulation of rodent PAs. Further, we show examples of experiments that can be performed with these arterioles, including agonist-induced constriction and myogenic reactivity. Although the focus of this manuscript is on PA cannulation and pressure myography, isolated PAs can also be used for biochemical, biophysical, molecular, and imaging studies.

This manuscript describes a simple and reproducible protocol for isolation of intracerebral arterioles (a group of blood vessels encompassing parenchymal arterioles, penetrating arterioles and pre-capillary arterioles) from mice, to be used in pressure myography, immunofluorescence, biochemistry, and molecular studies.

Intracerebral parenchymal arterioles (PAs), which include parenchymal arterioles, penetrating arterioles and pre-capillary arterioles, are high resistance ...

Cannulation of the Carotid Artery in a Rat Model

Transcript