AhR-KO (Ahr−/−) mice (B6.129-Ahrtm1Bra/J) [55 (link)] were purchased from Jackson Laboratories (JAX stock #002831) and maintained as a breeding colony in the animal care facility at the Faculty of Medicine of Marseille. C57BL/6J wild-type (WT) mice were used as experimental controls. Genotypes were confirmed by PCR analysis of DNA from tail clippings. Comparisons were made between sex-and age-matched groups. Kidney failure was induced in mice (10 weeks of age) by alternating a 0.25% adenine-enriched diet (A04 + 0.25% adenine; SAFE, Augy, France) and regular diet (A04 standard, SAFE) every other week for 6 weeks. Mice were sacrificed at 16 weeks after induction of CKD. Control mice were fed with a regular chow diet for 6 weeks. The experiments were performed in compliance with the Directive 2010/63/EU of the European Parliament and were approved by the local Ethics Committee (“Comité d’Ethique en Expérimentation Animale de Marseille”, C2EA-14; ethics approval number: 2017091414144363 V6; approval date: 20 November 2018).
B6.129 ahrtm1bra j
Manufactured by Jackson ImmunoResearch
Sourced in Montenegro, United States
B6.129-Ahrtm1Bra/J is a mouse strain with a targeted mutation in the Ahr gene. The Ahr gene encodes the aryl hydrocarbon receptor (AhR), which is a ligand-activated transcription factor that mediates the biological and toxicological effects of various environmental contaminants. This mouse strain can be used for research related to the role of the AhR in various physiological and pathological processes.
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7 protocols using b6.129 ahrtm1bra j
1
Adenine-Induced Kidney Failure in AhR-KO Mice
2
AhR Knockout Mice Protocol
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C57Bl/6 (H-2b/b), B6D2F1 (H-2b/d) and B6.129-AHRtm1Bra/J (AhR−/−) mice were obtained from The Jackson Laboratory. The AhR−/− mice were bred and maintained in a specific pathogen-free vivarium at Oregon State University (Corvallis, OR). OSU is an AALAC-accredited institution. Ethics Statement: All experimental procedures and treatments using animals (mice) were approved by the Institutional Animal Care and Use Committee at Oregon State University. Euthanasia was by CO2 overdose followed by cervical dislocation.
3
Investigating AhR Signaling in Intestinal Inflammation
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All reagents were from Sigma-Aldrich (Milan, Italy) unless specified. Eight week-old female C57BL/6J wild type (WT) and AhR knockout (KO) mice (B6.129-Ahrtm1Bra/J, Jackson Laboratories, Bar Harbor, ME) were given intra-peritoneally polyinosinic:polycytidylic acid (poly I:C) (15 μg/g) dissolved in phosphate buffered saline (PBS) or PBS only (controls) and sacrificed 12 h later through cervical dislocation. One hour after poly I:C administration, mice were given Ficz (1 μg/mouse). This dose was selected based upon our previous studies, which documented activation of AhR in the intestinal mucosa following i.p. administration of the compound (22 (link)). Small intestine was harvested for histology, RNA extraction, LPMC, and IEL isolation. The murine experiments were approved by the local Institutional Animal Care and Use Committee of the University of Tor Vergata.
4
Mouse Strains for Research
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C57BL/6J, ahr knockout mice (B6;129-Ahrtm1Bra/J), and p53 knockout mice (B6.129S2-Trp53tm1Tyj/J) were purchased from the Jackson Laboratory. CD1-IGS mice also called Crl:CD1(ICR) were purchased from Charles River. Mice were acclimated for at least 1 week following shipment and before experiments. Animal handling and experimental procedures were in accordance with the Guide for the Care and Use of Laboratory Animals and approved by the Emory University Institutional Animal Care and Use Committee.
5
Sciatic Nerve Explant Culture with AhR
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We obtained C57BL/6 mice (4–5 weeks old, 20–22 g) from the National Applied Research Laboratories (NAR labs, Taipei, Taiwan), and AhR-knockout mice (B6.129Ahrtm1Bra/J) (6–8 weeks old, 18–22 g) from the Jackson laboratory (Bar Harbor, ME, USA). The sciatic nerve was first harvested from both wild-type and AhR (−/−) C57BL/6J mice, and then 3 mm-thick nerve sections were cultured in calcium/magnesium-free Hank’s buffered solution (HBSS). Under a microscope, the nerve tissue was isolated from the surrounding connective tissues, and only single nerve bundles were analyzed. To determine the effects of AhR agonist, at doses on the explant nerves, omeprazole (doses from 0 to 40 μM) was added to the sciatic nerve cultures. The culture medium was DMEM with 10% FBS and 5% PS, at the condition of 5% CO2 and 37 °C for 1 to 7 days. Finally, nerve tissues were fixed in 4% paraformaldehyde for 6 h, and observed under a light microscope. The ovoid number in each 500 μm length of nerve was calculated for statistical analyses. The nerve explant cultures were categorized into Wild type-sham, Wild type-CCI, AhR (−/−)-CCI, Wild type-CCI-omeprazole, and AhR (−/−)-CCI-omeprazole.
6
Characterization of AhR-/- Mice
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Mice used included AhR−/− mice (B6.129-AhR^tm1Bra/J) 28 (link) originally obtained from Jackson Laboratory (Bar Harbour, ME, USA), where they were bred for 10 generations to C57BL6 mice and an additional two generations to C57BL/6 J mice. At Duke University, within the Division of Laboratory Animal Resources, over the last several years they have been bred a minimum of six generations to the 6 J background. Additionally, the mice were screened for the confounding retinal degeneration 8 mutation and its absence was confirmed as previously described 24 (link). Our study protocol was approved by the Duke University Institutional Animal Care and Use Committee. All animal experiments were performed in accordance with the guidelines of the ARVO statement for the Use of Animals in Ophthalmic and Vision Research.
7
Murine Model for Aryl Hydrocarbon Receptor Study
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Eight- to 12-week-old male C57BL6/J WT and B6.129-Ahrtm1Bra/J (AhR−/− mice)69 (link) from Jackson Laboratories, originally provided by Dr. Marc Veldhoen and bred as specific pathogen free mice at the Isogenic Breeding Unit of the Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, were used throughout this study.
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