Generation of Mouse Proximal Colon Loop Model: A Surgical Procedure to Generate Proximal Colon Loop and Measure Intestinal Permeability by Detection of Fluorescent Markers in Serum Following Intraluminal Injection

Abstract

Source: Boerner, K. et al. Functional Assessment of Intestinal Permeability and Neutrophil Transepithelial Migration in Mice using a Standardized Intestinal Loop Model. J. Vis. Exp. (2021).

In this video, we describe a surgical procedure to generate mouse proximal colon loop or pcLoop segment followed by intraluminal injection of fluorescent markers into the segment to study the intestinal epithelial permeability.

Protocol

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

1. Generation of the proximal colon loop (pcLoop)

1. Skin preparation: Scrub fur of the abdominal midline with alcohol swabs or gauze sponge soaked with 70% Ethanol. Do not wet a wide area of fur with alcohol to prevent hypothermia.
2. Using scissors, perform a midline laparotomy. Make a vertical incision in the middle of the abdomen (about 2 cm in length) and expose the peritoneum. Be careful to not injure intra-abdominal organs.
3. Place pre-cut wet cotton ....

Results

Figure 1: The proximal colon loop model. (A) Schematic overview of the pcLoop model. Median laparotomy is performed on mice under anesthesia placed on a te.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Equipment and Material
BD Alcohol Swabs	BD	326895
BD 1ml Tuberculin Syringe Without Needle	BD	309659
BD PrecisionGlide Needle, 25G X 5/8"	BD	305122
BD PrecisionGlide Needle, 30G X 1/2"	BD	305106
Cotton Tip Applicator (cotton swab), 6", sterile	FisherScientific	25806 2WC
Dynarex Cotton Filled Gauze Sponges, Non-Sterile, 2" x 2"	Medex	3249-1
EZ-7000 anesthesia vaporizer (Classic System, including heating units)	E-Z Systems	EZ-7000
Kimwipes, small (tissue wipe)	FisherScientific	06-666
Moria Fine Scissors	FST	14370-22
Puralube Vet Ointment, Sterile Ocular Lubricant	Dechra	12920060
Ring Forceps (blunt tissue forceps)	FST	11103-09
Roboz Surgical 4-0 Silk Black Braided, 100 YD	FisherScientific	NC9452680
Semken Forceps (anatomical<br/> forceps)	FST	1108-13
Sofsilk Nonabsorbable Coated Black Suture Braided Silk Size 3-0, 18", Needle 19mm length 3/8 circle reverse cutting	HenrySchein	SS694
Student Fine Forceps, Angled	FST	91110-10
10ml Syringe PP/PE without needle	Millipore Sigma	Z248029
Reagents
Isoflurane&nbsp;	Halocarbon&nbsp;	12164-002-25
Fluorescein Isothiocyanate-Dextran, average molecular weight 4.000	Sigma	60842-46-8