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

Published: April 30, 2023

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) 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. Using scissors, perform a midline laparotomy. Make a vertical incision in the middle…

Representative 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 temperature-controlled surgery board. (B) Exteriorization of the caecum (*), proximal colon, mesocolon and ileum. Two adequate sites for ligation are identified (1,2). (C) The first ligature (1) …

Açıklamalar

The authors have nothing to disclose.

Materials

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
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  Halocarbon  12164-002-25
Fluorescein Isothiocyanate-Dextran, average molecular weight 4.000 Sigma 60842-46-8

Etiketler

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Bu Makaleden Alıntı Yapın
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. J. Vis. Exp. (Pending Publication), e20813, doi: (2023).

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