intraduodenal cannulation in a rat model for immunopathology studies

Intraduodenal Cannulation in a Rat Model for Immunopathology Studies

To cannulate the duodenum, first, locate the tissue as the bright pink section of the small intestine, which upon gentle downward pulling reveals the stomach. Then, use a sterile 23-gauge needle to make a small puncture hole in the duodenum at the pylorus, approximately 2 centimeters below the junction of the stomach and duodenum. Next, insert the J-shaped hook end of the duodenal cannula through the puncture and secure the cannula in place with a drop of cyanoacrylate glue. Then, use sutures to close the abdominal muscle wall incision.

intraduodenal-cannulation-in-a-rat-model-for-immunopathology-studies

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JoVE Encyclopedia of Experiments

Immunology

Immunopathology

Assay and Analysis

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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. Preparations the Day Before the Surgical Procedure
<ol>
	<li>Fast the rat the night before surgery if required. Ensure the rat has free access to water.</li>
	<li>Prepare the solutions to be administered to the rat.
	<ol>
		<li>Prepare a rehydration solution such as sterile saline or Ringers solution.</li>
		<li>Prepare anesthetic solutions as required. The anesthetic used in the experiments described here consisted of&#160; &#34;Cocktail 1&#34; prepared by combining 1.9 ml of ketamine 100 mg/ml, 0.5 ml of Xylazine 100 mg/ml, 0.2 ml of acepromazine 10 mg/ml, and 2.5 ml of saline and &#34;Cocktail 2&#34; consisting of 1 ml of ketamine 100 mg/ml and 0.1 ml acepromazine 10 mg/ml. NOTE: Inhaled isoflurane or sevoflurane may be preferred as it can provide a surgical plane of anesthesia for a longer period of time.</li>
	</ol>
	</li>
	<li>Prepare an anti-coagulant solution containing 10 mg/ml ethylenediaminetetraacetic acid (EDTA) in sterile water or 10 IU/ml heparin in saline to flush the lymph cannula. Also, prepare an anti-coagulant solution containing 2.5 - 10 IU/ml heparin in saline to flush the carotid artery cannula.</li>
	<li>Prepare the polyethylene cannula for insertion into the duodenum (0.96 mm O.D, 0.58 mm I.D.)
	<ol>
		<li>Cut the cannula to the required length (typically 25 - 30 cm for the duodenal cannula) and place a bevel at the tip of the cannula using a sterile surgical blade (see&#160;Figure 1).&#160;&#160;&#160; 
		NOTE: This increases the ease of cannula insertion and reduces the incidence of cannula blockage post-insertion.</li>
		<li>Place a J-shaped anchor point in the intraduodenal cannula by looping a small portion of the cannula back on itself, heating it with a lighter or hot plate, and then cutting it to size (see&#160;Figure 1).</li>
	</ol>
	</li>
	<li>Attach the lymph cannula to a 25 G needle and syringe containing an anti-coagulant solution (prepared in step 1.3). Fill the lymph cannula with the anti-coagulant solution and leave the solution in the cannula overnight (O/N) to reduce the incidence of clot formation in the lymph cannula during lymph collection.</li>
	<li>Prepare tubes for lymph and blood sample collection. Pre-weigh lymph collection tubes, label tubes for lymph and blood sample collection and add anti-coagulant solution. Add sufficient anti-coagulant solution to achieve a final concentration of 1 mg/ml ethylenediaminetetraacetic acid (EDTA) in sterile water or 10 - 20 IU/ml heparin in the collected lymph or blood.</li>
</ol>
2. Preparations Immediately Prior to Commencing the Surgical Procedure
<ol>
	<li>Check the cannulas by flushing them with sterile saline or anti-coagulant solution to ensure that they are patent.</li>
	<li>Prepare the rat for the surgical procedure.
	<ol>
		<li>Anesthetize the rat throughout the surgery and sample collection period.
		<ol>
			<li>For example, initiate the anesthesia via subcutaneous injection of 1.5 ml/kg of Cocktail I (from step 1.2.2) into a skin fold at the back of the rat neck using a 1 ml syringe attached to a 25 G needle. Maintain the anesthesia via intraperitoneal injection of&#160; 0.44 ml/kg of Cocktail 2 (from step 1.2.2) approximately each hour as required, using a 1 ml syringe attached to a 25 G needle.</li>
			<li>Prior to commencing the surgery, ensure that the depth of anesthesia is sufficient by observing respiratory rate, whisker movement, muscle tone, and responses to stimuli such as pinching of the foot. Administer additional anesthesia as required.</li>
		</ol>
		</li>
		<li>Shave the fur from the surgical regions, which include the right side of the abdomen, for duodenum cannulation.</li>
		<li>Clean the surgical regions aseptically using povidone-iodine solution or chlorhexidine solution and a 70% ethanol scrub. Repeat this 3 times for each region, finishing with a final cleansing with 70% ethanol.</li>
	</ol>
	</li>
	<li>Place the animal in dorsal recumbence on a clean sheet above a heated surgical pad (37 &#176;C).</li>
</ol>
3. Cannulation of the Duodenum
<ol>
	<li>Place the animal with its right side facing toward the operator. Perform the surgery with or without the assistance of a surgical microscope as required.</li>
	<li>Open the top layer of the abdominal muscle wall with a straight 4 cm incision extending from the midline (xiphoid process) to the right flank approximately 2 cm below the ribcage (costal margin) using a sterile scalpel blade (see Figure 2B).</li>
	<li>Open the remaining layers of the abdominal muscle wall from 4 - 5 mm lateral to the midline to the right flank with a small pair of surgical scissors (see Figure 2B).</li>
	<li>Retract the small intestine under the left abdominal muscle wall and keep it in place using 2 - 3 pieces of sterile gauze saturated with normal saline.</li>
	<li>Attach the duodenum cannula to a syringe (e.g.,&#160;10 ml) filled with the rehydration solution.</li>
	<li>Identify the duodenum as the bright pink (with more blood vessels) section of the small intestine, which upon gentle downward pulling reveals the stomach.</li>
	<li>Make a small puncture hole in the duodenum approximately 2 cm below the junction of the stomach and duodenum (the pylorus) using a sterile 23 G needle.</li>
	<li>Insert the J-shaped hooked end of the duodenal cannula through the puncture hole. Secure in place with a drop of cyanoacrylate glue.</li>
	<li>Commence hydration of the rat by attaching the cannula to the rehydration syringe loaded in an infusion pump. According to the literature, rehydration rates range from 0.5 - 3 ml/hr.</li>
	<li>After completion of the lymph and duodenum cannulation, close the incision in the abdominal muscle wall with sutures. Then close the skin incision by applying a few drops of tissue adhesive (cyanoacrylate) along the side of the incision and pinching the skin on both sides of the incision together.</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>Chlorhexidine gluconate solution (Microshield 4)</td>
			<td>Livingstone International</td>
			<td>JJ60243L</td>
			<td>Any brand can be used. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search</td>
		</tr>
		<tr>
			<td>Betadine solution</td>
			<td>Livingstone International</td>
			<td>BU0510</td>
			<td>Any brand can be used. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search</td>
		</tr>
		<tr>
			<td>Ilium Ketamil (Ketamine 100 mg/ml)</td>
			<td>PROVET VICTORIA&#160;</td>
			<td>KETA I 1</td>
			<td>http://www.provet.com.au/</td>
		</tr>
		<tr>
			<td>Ilium Xylazil (Xylazine 100 mg/ml)</td>
			<td>PROVET VICTORIA&#160;</td>
			<td>TRO-3828</td>
			<td>http://www.provet.com.au/</td>
		</tr>
		<tr>
			<td>ACP 10 Injection (Acepromazine 10 mg/ml)</td>
			<td>PROVET VICTORIA&#160;</td>
			<td>VTG-DACP010020</td>
			<td>http://www.provet.com.au/</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>Ethylenediaminetetraacetic acid (EDTA) disodium salt dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>E1644</td>
			<td>Any brand can be used. Example here is disodium salt of EDTA from Sigma.&#160;</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>Cyanoacrylate glue</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&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; =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&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; =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&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; =GSC075</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Ratek</td>
			<td>WT1</td>
			<td>Any brand that keeps temperature at 37C can be used. Example here is Ratek warming tray.</td>
		</tr>
		<tr>
			<td>Surgical light</td>
			<td>Harvard Apparatus</td>
			<td>72-0215 with 72-0267</td>
			<td>Any brand can be used. Example here is Harvard apparatus V-Lux 1000 Cold Light Source with Bifurcated Gooseneck Light Guide, Black, 4.7 mm fiber diameter (each arm). http://www.harvardapparatus.com/webapp/wcs/stores/servlet/product_11051_10001_50601_</td>
		</tr>
		<tr>
			<td>Surgical microscope</td>
			<td>Zeiss</td>
			<td>495005-0014-000</td>
			<td>Any brand can be used. Example here is Zeiss Stereomicroscope Stemi 2000-C with Stand S Double Spot and KL 300 LED. https://www.micro-shop.zeiss.com/?l=en&#38;p=us&#38;f=e&#38;i=10143</td>
		</tr>
		<tr>
			<td>Silk suture</td>
			<td>Livingstone International</td>
			<td>DTSK163019F4</td>
			<td>Any brand can be used. Example here is *&#160;Email this item to my friend&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 3/8 Circle Reverse Cut Silk Suture 3/0 Thread 19mm. http://www.livingstone.com.au/?PG=search_result&#38;CAT=6&#38;search&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; =DTSK163019F4</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>2 x Hemostats</td>
			<td>Fine Science Tools (FST)</td>
			<td>13010-12</td>
			<td>Any brand can be used. Not all operators use the hemostats. Example here is FST 12 cm Micro-Mosquito Hemostats with 20 mm length x 1.3 mm width serrated, straight tip. http://www.finescience.ca/Special-Pages/Products.aspx?ProductId=377&#38;CategoryId=33</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>
	</tbody>
</table>

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>&#160;J. Vis. Exp.&#160;(2015)
This video demonstrates intraduodenal cannulation in a rat model. After anesthetizing the rat, the duodenum is located and a small puncture is created. A J-shaped cannula with a beveled tip is carefully inserted and secured, and the incision is closed. The cannulated rat model enables intra-intestinal infusion for immunopathology studies.

<img alt="Figure 1" src="/files/ftp_upload/21760/21760_Figure1.jpg" /> 
Figure 1. Diagrammatic representation of the shape of the beveled tip of a carotid artery or lymph cannula, and the J-shaped beveled tip of a duodenal cannula.
<img alt="Figure 2" src="/files/ftp_upload/21760/21760_Figure2.jpg" /> 
Figure 2. Photographs of (A) the abdomen shaved in preparation to cannulate the lymp...

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 rat in a supine position.
Shave the abdominal region and dissect the rat laterally, involving the skin and muscular layer to access the abdominal organs.
Visualize the duodenum&#160; &#8212; a C-shaped tube connecting the stomach to the small intestine&#39;s middle part &#8212; the jejunum.
Pull the duodenum downward to expose the stomach and locate the pylorus&#160; &#8212; the stomach-duodenum junction.
Identify the proximal region below the pylorus and create a small puncture in the duodenum.
Take a J-shaped cannula&#160; &#8212; a thin tube with a beveled tip for smoother insertion, and avoid dislocation.
Insert the cannula through the duodenal puncture to access the intra-intestinal cavity and secure its entry point on the duodenum.
Post-cannulation, suture the muscular layer incision, ensuring the cannula&#39;s open end remains outside the rat&#39;s body, and finally, apply surgical glue to close the skin incision.
The intraduodenal cannulated rat model is ready for intra-intestinal infusion of substances for immunopathology studies.

320 vues. Canulation intraduodénale dans un modèle de rat pour des études d’immunopathologie

Vidéo: Canulation intraduodénale dans un modèle de rat pour des études d’immunopathologie - Expérience

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

Implantation de la canule dans la Magna de Cisterna de rongeurs

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

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Neurosciences

Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, which could potentially affect results. The jugular vein cannulation (JVC) in rats is a widely used model for repeated blood collection but requires adequate training of surgery skills and animal care. This article details the microsurgical procedures for establishing and maintaining a permanent JVC rat model with specific focus on the placement and sealing of the jugular cannula. The importance of monitoring physiological (e.g., body weight, food, and water intake) and hematological parameters, was highlighted with results presented for 6 days post-surgery during the rat&#39;s recovery. The drug-plasma concentration-time profile of orally administered natural phenol ellagic acid was determined in the JVC rat model.

Detailed microsurgical techniques are demonstrated to establish a longer-term jugular vein cannulation rat model for sequential blood collection in the same animal. Physiological and hematological parameters have been monitored during the rat's recovery phase. This model has been applied to study pharmacokinetics of orally administered polyphenol without inducing animal stress.

Compétences microchirurgicales pour établir une canulation veineuse jugulaire permanente chez le rat pour le prélèvement sanguin en série d’un médicament administré par voie orale

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, ...

Médecine

Fully Endoscopic Mitral Valve Repair with Percutaneous Cannulation of Groin Vessels

Endoscopic mitral valve surgery (EMS) has become a standard of care at specialized heart centers, further reducing surgical trauma compared to a traditional minimally invasive, thoracotomy-based approach. Exposure of the groin vessels for the establishment of cardiopulmonary bypass (CPB) via surgical cutdown in minimally invasive surgery (MIS) may result in wound healing disorders or seroma formation. The avoidance of surgical exposure of the groin vessels by using fully percutaneous techniques for the insertion of a CPB cannula with the implementation of vascular pre-closure devices has the potential to reduce these complications and improve clinical results. Herein, we present the utilization of a novel plug based vacsular closure device with a resobable collagen plug and the absence of suture material for closure of the arterial access for CPB in MIS. While this device was initially predominantly used in transcatheter aortic valve implantation (TAVI) procedures, with its safety and feasibility shown, we herein show that it can be used in CPB cannulation, since it is capable of closing arterial access sites up to 25 French (Fr.) in size. This device may be suitable to significantly reduce groin complications in MIS and simplify the establishment of CPB. Here, we describe the fundamental steps of EMS, including percutaneous groin cannulation and decannulation using a vascular closure device.

This protocol demonstrates in detail how to perform fully endoscopic mitral valve surgery (EMS) with percutaneous cannulation of the groin vessels, using a percutaneous plug-based vascular closure device. Fundamental steps and useful instructions are described in detail for each step.

Intraduodenal Cannulation in a Rat Model for Immunopathology Studies

Transcription

Intraduodenal Cannulation in a Rat Model for Immunopathology Studies