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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. General Preoperative Preparation
<ol>
	<li>Fast the mice for 6 h before the surgery. Gel diet food was given 3 days before surgery and exclusively one day before surgery. Induce anaesthesia in a chamber with 5% isoflurane (0.4 L/min) and O2&#160;(dioxygen; 0.4 L/min). Perform a toe-pinch test to confirm that the anesthetization is effective. Administer analgesics (0.1 mg/kg buprenorphine), antibiotics (10 mg/kg marbofloxacin), and pro-kinetics (1 mg/kg metoclopramide)&#160;via&#160;subcutaneous injections.</li>
	<li>Allocate a specific work area for surgical procedures (distant from laboratory traffic and ventilation fans). Clean the surgical area with dedicated hard surface disinfectants. Place a heating pad on the working space area to maintain mouse homeothermy during the surgery. Apply a clean, absorbent under pad over the heating pad.</li>
	<li>Shave the abdomen from sternum to pelvis using an electric razor. Wrap the mouse abdomen in sterilized plastic wrap (Figure 1A) and place the mouse in supine position on the heating pad.</li>
	<li>Apply vet ointment on the eyes to prevent dryness while under anaesthesia. Place the mouse in the nose cone and maintain anaesthesia with 2% isoflurane (0.4 L/min) and O2&#160;(0.4 L/min).</li>
	<li>Before beginning the surgery, put on a sterile examination gown, a disposable scrubs cap, a facemask, and sterile gloves. Open an autoclaved package containing a sterilized surgical instrument set, which is required for all steps of the surgery. Cut a window in the sterilized plastic wrap and disinfect the skin with 2 successive povidone-iodine solutions.</li>
</ol>
2. Roux-en-Y Gastric Bypass: Surgical Procedure
<ol>
	<li>Median laparotomy
	<ol>
		<li>Under a binocular microscope and with a scalpel, perform a midline incision from the sternum to the middle of the abdomen to open the abdominal cavity (do not damage the abdominal muscles). Protect the skin with a sterile compress soaked with 37 &#176;C saline solution (Figure 1B).</li>
	</ol>
	</li>
	<li>Biliopancreatic limb and alimentary limb
	<ol>
		<li>Externalize the intestine (Figure 1C). Measure 8 cm from the pylorus and perform two ligatures of the intestine with 5.0 non-absorbable sutures&#160;(Figure 1D).</li>
		<li>Cut the intestine between the two ligatures (Figure 1E). Place the proximal limb of the two ends in the upper-left quadrant of the abdomen. Note: This will be used as the alimentary limb (Figure 1F).</li>
		<li>Place the distal limb of the two ends facing the alimentary limb 6 cm below the proximal limb (Figure 1F). 
		NOTE: This will be used as the biliary limb.</li>
	</ol>
	</li>
	<li>Jejuno-jejunostomy
	<ol>
		<li>Cut both the proximal limb and the intestine loop using micro scissors and perform two antimesenteric incisions of the same length (Figure 1G and 1H).</li>
		<li>Perform side-to-side anastomosis with two 8.0 non-absorbable sutures. Perform the dorsal side anastomosis first (Figure 1I), followed by the ventral side anastomosis (Figure 1J).</li>
		<li>Gently roll 2 moistened cotton swabs towards each side of the anastomosis to ensure that the suture is leak-proof. Complete with 8.0 non-absorbable sutures in case of leakage.</li>
	</ol>
	</li>
	<li>Gastro-jejunostomy
	<ol>
		<li>Gently mobilize the stomach using moistened cotton swabs. Free the stomach from its lateral close connective tissue attachments using moistened cotton swabs or micro scissors.</li>
		<li>Gently, externalize the stomach fully, placing a re-absorbable hemostatic collagen compress behind the stomach. Perform a ligature of the pylorus using a 5.0 non-absorbable suture passed through the omentum using curved micro forceps (Figure 1K&#160;and&#160;1L).</li>
		<li>Cut both the ventral side of the stomach, 1.5 cm from the pylorus, and the distal limb using micro scissors, creating two incisions of the same length (Figure 1M).</li>
		<li>Perform side-to-side anastomosis with two 8.0 non-absorbable sutures. Start with the dorsal side anastomosis (Figure 1N), and then perform the ventral side anastomosis (Figure 1O). Gently roll 2 moistened-cotton swabs towards each side of the anastomosis to ensure that the suture is leak-proof. Complete with 8.0 non-absorbable sutures in case of leakage.</li>
	</ol>
	</li>
	<li>Abdominal closure
	<ol>
		<li>Close the muscle layer of the abdominal wall using 5.0 non-absorbable sutures (Figure 1P). Reduce the anaesthesia by reducing the isoflurane concentration to 1%. Close the skin using 5.0 non-absorbable sutures (Figure 1Q). Administer 25 mL/kg of warm saline solution via subcutaneous injection.</li>
	</ol>
	</li>
</ol>
3. General Postoperative Care
<ol>
	<li>Stop isoflurane and continue with an O2&#160;flow of 0.8 L/min until the mouse is fully awake. Do not leave the mouse unattended until it regains motor control, indicated by the animal starting to move around the cage and being able to stand and walk without falling.</li>
	<li>Place the mouse (only one mouse per cage) in an incubator under 30 &#176;C temperature condition for 5 days.</li>
	<li>Maintain iron (0.5 mg/kg/day; subcutaneous injection) and vitamin (800 mg/180 mL in water) supplementations in the RYGB mice until the end of the protocol.</li>
	<li>Return free access to gel diet food (high-fat gel diet: 10% lard, 10% liquid sugar, 57% water) for 5 days after surgery. Reintroduce a solid diet 3 days after surgery.</li>
	<li>Subcutaneously inject buprenorphine (0.1 mg/kg, twice daily, from day 0 to day 3 after surgery), meloxicam (1 mg/kg, from day 0 to day 3 after surgery), metoclopramide (1 mg/kg, from day 0 to day 5 after surgery), and marbofloxacin (10 mg/kg, from day 0 to day 3 after surgery)</li>
</ol>

<table><tbody><tr><td>Drugs</td><td></td><td></td><td></td></tr><tr><td>High Fat diet</td><td>DIO diet</td><td>Safe</td><td></td></tr><tr><td>Isoflurane</td><td>Forane</td><td>Baxter</td><td></td></tr><tr><td>Buprenorphin</td><td>Buprecare</td><td>Animalcare</td><td></td></tr><tr><td>Marbofloxacine</td><td>Marbocyl</td><td>Vetoquinol</td><td></td></tr><tr><td>Ammonium iron citrate, vitamins&lt;br/&gt; PP-B12</td><td>Fercobsang</td><td>Vetoquinol</td><td></td></tr><tr><td>Vitamins A-D3-E-K-B</td><td>Vita Rongeur</td><td>Virbac</td><td></td></tr><tr><td>NaCl 0.9%</td><td>NaCl 0,9%</td><td></td><td></td></tr><tr><td>Povidone solution</td><td>Betadine Scrub</td><td>Betadine</td><td></td></tr><tr><td>Povidone solution</td><td>Betadine Solution</td><td>Betadine</td><td></td></tr><tr><td>Carboptol 980 NF</td><td>Ocrygel</td><td>TVM</td><td></td></tr><tr><td>Sutures</td><td></td><td></td><td></td></tr><tr><td>Prolene&amp;reg;</td><td>8.0, 6,5 mm</td><td>Ethicon</td><td></td></tr><tr><td>Prolene&amp;reg;</td><td>5.0, 13 mm</td><td>Ethicon</td><td></td></tr><tr><td>Surgical equipments</td><td></td><td></td><td></td></tr><tr><td>Scissors</td><td></td><td>FST</td><td></td></tr><tr><td>Needle holder</td><td>Olsen-Hegar</td><td>FST</td><td></td></tr><tr><td>Micro scissors</td><td>Vannas</td><td>FST</td><td></td></tr><tr><td>Micro forceps</td><td>Graefe</td><td>FST</td><td></td></tr><tr><td>Micro forceps curved</td><td>Graefe</td><td>FST</td><td></td></tr><tr><td>Curved micro needle holder</td><td>Castroviejo</td><td>FST</td><td></td></tr><tr><td>Hemostatic collagen compress</td><td>Pangen</td><td>Urgo</td><td></td></tr><tr><td>Absorbent underpads</td><td></td><td>VWR</td><td></td></tr><tr><td>Specific equipments</td><td></td><td></td><td></td></tr><tr><td>Stereo microscope</td><td>MZ6</td><td>Leica</td><td></td></tr></tbody></table>

roux-en-y gastric bypass procedure: a bariatric surgical procedure for gastric resectioning in murine model

Source: Ayer, A., et al., <a href="https://jove.unicartagenaproxy.elogim.com/v/54905">Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice.</a> J. Vis. Exp. (2017).
This video describes a bariatric surgery-Roux-en-Y procedure. This involves the cutting of the jejunum- the middle portion of the intestine- to generate two intestinal limbs. The biliary limb is attached to the existing intestinal loop and the alimentary limb is attached to the stomach, creating a gastric bypass. The gastric bypass created here helps to decrease the nutrient uptake and efficiently reduces body weight.

<img alt="Figure 1" src="/files/ftp_upload/20815/20815_Figure1.jpg" /> 
Figure 1: Roux-en-Y bypass Procedure.&#160;(A) Preoperative preparation. (B) Median laparotomy. (C) Externalization of the small intestine. (D) Biliopancreatic and alimentary limbs ligatures. (E) Small intestine section between the ligatures. (F) Intestinal limb positioning for the jejuno-jejunos...

Roux-en-Y Gastric Bypass Procedure: A Bariatric Surgical Procedure for Gastric Resectioning in Murine Model

Source: Ayer, A., et al., Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice. J. Vis. Exp. (2017).
This video describes a ...

Begin the Roux-en-Y procedure &#8212; a gastric resection surgery &#8212; by placing an anesthetized mouse in the supine position. Dissect the abdomen and externalize the small intestine to locate the jejunum &#8212; the middle part of the intestine.
Perform two nearby ligations on the jejunum and make an excision between them. This process generates two intestinal limbs &#8212; the biliary limb is proximal to the stomach and the alimentary limb is distal to the stomach. Place the biliary limb near the existing intestinal loop. Make equal-sized longitudinal incisions on their antimesenteric surface.
Perform side-to-side anastomosis by suturing incised regions of the biliary limb and the intestinal loop, allowing the gastric juices to enter intestines directly. Locate the stomach and ligate the pylorus &#8212; the portion of the stomach that opens into the intestine, which prevents the food from entering the intestine.
Make a new longitudinal incision near the pylorus and the pulled end of the alimentary limb and perform the second side-to-side anastomosis. This new path is called the Roux limb, and the intestine now resembles &#8216;Y&#8217; shape. Return the ligated organs to the abdominal cavity and suture the incision site.
Post-surgery, the food directly enters from the stomach to the Roux limb, bypassing the normal gastric route. The smaller intestinal length decreases nutrient absorption, causing a reduction in the mouse&#8217;s weight.

roux-en-y-gastric-bypass-procedure-bariatric-surgical-procedure-for

Universidad de Cartagena UDEC

Research

JoVE Encyclopedia of Experiments

Biology

Rodent Models

Adult

1.2K ビュー. 出典:Ayer, A., et al., Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice. J. Vis. Exp. (2017). このビデオでは、肥満手術-Roux-en-Yの手順について説明しています。これは、腸の中央部である空腸を切開して、腸の手足を2つ作り出すものです。胆道肢は既存の腸ループに付着し、消化肢は胃に付着して胃バイパスを作成します。ここで作られた胃バイパスは、栄養素の摂取?...

ビデオ: Roux-en-Y胃バイパス術:マウスモデルにおける胃切除術のための肥満外科手術 - 概念

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

ラットにおけるルー-en-Y法胃バイパス手術

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

JoVE Journal

医学

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

リニア・ステープラーを用いたラットにおけるルー・アン・Y胃バイパスのプロトコル

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Robotic Central Pancreatectomy with Roux-en-Y Pancreaticojejunostomy

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of the pancreas. The aim of central pancreatectomy is to prevent postoperative life-long endocrine and exocrine insufficiency. The downside of central pancreatectomy is the high rate of postoperative pancreatic fistula, which is the main reason that many surgeons do not routinely use central pancreatectomy in eligible patients. Most studies report open or laparoscopic central pancreatectomy with a pancreatico-gastrostomy anastomosis in adults. This is the first description of a standardized approach to robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy reconstruction in an adolescent (16-year-old boy) with a pseudopapillary tumor in the body of the pancreas. The operation time was 248 min with 20 mL of blood loss. The postoperative course was uneventful except for the short-term medical treatment for a grade B pancreatic fistula. Robotic central pancreatectomy can be safely applied in selected patients in experienced centers.

Robotic central pancreatectomy may be used in selected patients in experienced centers. This protocol presents all steps and the feasibility of a robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy in a 16-year-old adolescent patient.

ルー・アン・イ膵チコジュノミノストミーによるロボット中央汎創切術

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of ...

Roux-en-Y Gastric Bypass Procedure: A Bariatric Surgical Procedure for Gastric Resectioning in Murine Model

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Roux-en-Y Gastric Bypass Procedure: A Bariatric Surgical Procedure for Gastric Resectioning in Murine Model