Eight-week-old male C57BL/6J mice (Jackson Laboratories, Bar Harbor, ME) were used for behavioral pharmacology experiments. To visualize Pdyn-expressing neurons, we generated a Pdyn-GFP reporter line by crossing preprodynorphin-IRES-Cre mice (Crowley et al., 2016 (link); Bloodgood et al., 2020 (link); Al-Hasani et al., 2015 (link)) (PdynIRES-Cre, B6;129S-Pdyntm1.1(cre)Mjkr/LowlJ, Jackson Laboratories Stock # 027958) and Rosa26-flox-stop-L10a-EGFP reporter mice (EGFP-L10a; B6;129S4-Gt(ROSA)26Sortm9(EGFP/Rpl10a)Amc/J, Jackson Laboratories Stock # 024750). For conditional knockout of BNST Pdyn, we used the Pdynlox/lox mouse line (Bloodgood et al., 2020 (link)). These mice were bred in the UNC facilities. All mice were group-housed for at least 3 days before being singly housed in polycarbonate cages (GM500, Tecniplast, Italy) with a 12:12 hr reversed dark-light cycle with lights off at 7:00am. Mice had unrestricted access to food (Prolab Isopro RMH 3000, LabDiet, St. Louis, MO) and H2O. The UNC School of Medicine Institutional Animal Care and Use Committee approved all experiments. Procedures were conducted in accordance with the NIH Guidelines for the Care and Use of Laboratory Animals.
Gm500
Manufactured by Tecniplast
Sourced in Italy
The GM500 is a compact and versatile laboratory equipment designed for general use in research and testing environments. It provides a controlled environment for various applications. The core function of the GM500 is to maintain specific environmental conditions within an enclosed space.
Automatically generated - may contain errors
Lab products found in correlation
C57bl 6j mice, by Jackson ImmunoResearch (4 mentions)
Isopro rmh 3000, by LabDiet (2 mentions)
Prolab isopro rmh 3000, by LabDiet (1 mentions)
C57bl 6j, by Inotiv (1 mentions)
Balb c, by Inotiv (1 mentions)
C57bl 6j 000664, by Jackson ImmunoResearch (1 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Teklad global 18 protein rodent diet 2018 chow diet, by Inotiv (1 mentions)
C57bl 6j male mice, by Charles River Laboratories (1 mentions)
C57bl 6n mice, by Charles River Laboratories (1 mentions)
17 protocols using gm500
1
Genetic Manipulation of Pdyn-Expressing Neurons
2
Mice Strain Comparison for Research
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Mice from three distinct strains: BALBc, C57BL/6J, and CD-1 (ICR) were purchased from Envigo (Rehovot, Israel). The mice were kept in clean plastic chambers (GM500, Tecniplast, Italy) at 22°C and a 12-h light/12-h dark cycle (light on at 7 am) and received food and water ad libitum. All cages contained standard wood chip bedding, cotton wool bedding material, and a mock nest of a 12 cm plastic tube. All the recorded adult mice were 3–5 months old, and all the recorded pups were healthy, 3–5 days old pups.
3
Pneumococcal Infection Mouse Model
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Mice were housed at the University of Liverpool biomedical services unit at 21°C to 23°C with 55 to 65% humidity. Mice were placed into individually ventilated cages (GM500; Tecniplast). Automatic watering provided reverse-osmosis water sterilized by UV radiation. Enrichment included nesting material, a balcony, a dome home, and a handling tunnel. For virulence testing, 7- to 8-week-old female C57BL/6 mice were used, after 7 days of acclimatization to the animal unit. Mice were anesthetized with O2-isoflurane and infected intranasally with 1 × 106 CFU of S. pneumoniae in 50 μL of PBS. Mice were periodically scored for clinical signs of disease and culled when severity limits were reached or at predetermined times postinfection. Severity limits are based on a scoring system that considers animal appearance (hunching and changes in the coat), natural and provoked behavior (depressed or elevated activity), weight loss, and respiratory patterns. A moderate change across three or more categories, or a substantial change in any single category, is the defined severity limit. Blood samples were obtained through tail bleeds or cardiac punctures under terminal anesthesia.
4
Bdnf Knockout Mice for Behavioral Study
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Female adult mice (16–18 weeks old at the beginning of the experiments) of C57BL/6J-000664 background (from Jackson Laboratories, Bar Harbor, ME, USA, maintained in the Laboratory Animal Center of the University of Helsinki), carrying a deletion in one of the copies of Bdnf gene or wild-type littermates were used [17 (link)]. The animals were group housed (4–5/cage; type-II individually ventilated cages GM500, 391 × 199 × 160 mm, floor area 501 cm2; Tecniplast, Buguggiate, Varese, Italy) in a 12-h light/12-h dark cycle (light on at 7:00 a.m.), with free access to food and water except during the experimental sessions. All protocols were approved by the ethics committee for animal experimentation of Southern Finland (ESAVI/38503/2019).
5
Ethical Animal Experiments for Mice
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All animal experiments were performed according to the FELASA guidelines and upon ethical approval by the Regierung von Oberbayern including a priori group size estimations (effect size 2.0, alpha error 0.05, beta error 0.2) in order to prevent underpowering. Female C57BL/6 mice (8–10 weeks old, in total n = 29) were obtained from Charles River (Sulzfeld, Germany) and were housed in groups of 4 animals in individually ventilated cages (GM500, Tecniplast, Hohenpeißenberg, Germany) in a specified pathogen-free animal facility with a 12 h day/night cycle. Standard rodent feed (Ssniff, Soest, Germany) and water were provided ad libitum. Animals were inspected daily and were sacrificed when reaching a pre-defined health score comprising critical weight loss, neurological symptoms, and overall health performance.
6
In Vivo Assessment of Therapeutic Treatments
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All animal care and procedures were performed in compliance with the regulation on the use of Animals in Research (UK Animals Scientific Procedures Act of 1986 and the EU Directive 2010/63/EU) under the project license number P9DCDB3B0 and with approval from the LMB Animal Welfare and Ethical Review committee. C57BL/6J male mice were used for the assessment of treatment effects. Animal cohort numbers were determined by power analysis based on preliminary results or literature precedent. Mice were housed in pathogen-free ventilated cages (Tecniplast GM500, Tecniplast) on Lignocel FS14 spruce bedding (IPS) and Enviro-Dri nesting material (LBS) at 19–23 °C with 12 h light–dark cycle with light from 7.00 am to 7.00 pm. The number of cages were kept to between 2 to 3 animals per cage. Every day a visual and physical health check of all experimental animals was performed. The experimental animals were weighed prior to the start of the experiment.
7
Fmr1-KO Mouse Model for Fragile X Syndrome
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Mouse strain B6.129P2‐Fmr1tm1.2Cidz/J (also called Fmr1‐KO2 model, missing exon 1 (Mientjes et al, 2006 (link)) males from same genotype (thereafter called Fmr1‐KO or Fmr1‐WT) were grouped by 3 or 4 individuals at weaning age (4 weeks), in individually ventilated cages (GM500, Tecniplast, UK), with poplar shaving bedding (Lignocell Select, JRS, Germany), and maintained under standard conditions, on a 12‐h light/dark cycle (7 h/19 h), with standard diet food (standard diet D04, Scientific Animal Food and Engineering, France) and water available ad libitum. Mice from a same cage received the same treatment and were transferred in the animal facility of the phenotyping area the next week.
Animal work involved in this study was conducted according to ARRIVE guidelines and received authorization from relevant national committee (Comité National de Réflexion Ethique en Expérimentation Animale) with number APAFIS#5874‐20l6062915583967 v2 and APAFIS #17544‐2018111516571205 v7 at the ICS mouse facility (Illkirch, France). Every procedure on mice was performed with the aim of ensuring that discomfort, distress, pain, and injury would be minimal.
Animal work involved in this study was conducted according to ARRIVE guidelines and received authorization from relevant national committee (Comité National de Réflexion Ethique en Expérimentation Animale) with number APAFIS#5874‐20l6062915583967 v2 and APAFIS #17544‐2018111516571205 v7 at the ICS mouse facility (Illkirch, France). Every procedure on mice was performed with the aim of ensuring that discomfort, distress, pain, and injury would be minimal.
8
Alcohol Consumption in C57BL/6J Mice
9
Murine In Vivo Experiments: Standardized Protocols
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Adult female mice (8–15 weeks old) were used for the in vivo experiments. The wild-type C57BL/6 or FVB mice were obtained from Janvier (Le Genest-Saint-Isle, France). Mice were housed in a specific pathogen-free facility, in individually ventilated cages GM500 (Tecniplast) or rat R.BTM.U x/R.ICV.6 cages (Innovive, Paris, France) and single-sided rat Innoracks (cat# RS.5.8.40) containing corn cob bedding. Each cage contained no more than five mice. The animals were maintained in a controlled-temperature room (22 ± 1°C) under a 14 h light/10 h dark cycle. The environment was enriched with two pieces of wipes, one cardboard tunnel, one cardboard or polysulfone house with two entrances/exits. Food (SAFE® 150, Safe, Rosenberg, Germany) and water were available ad libitum. All experimental procedures were performed in strict accordance with Swiss regulations concerning the care and use of laboratory animals (veterinary authorizations 3447 and 3682).
10
High-Fat Diet Impacts Proximal Colon
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Mice were studied as previously described (Williams et al., 2016 (link)) and multiple organs were harvested for further analysis. Briefly, in groups of 3–5 animals from the same strain and diet, in isolator cages with individual air filtration (500 cm2, GM500, Tecniplast) and provided water ad libitum. Mice were fed CD ad libitum until 8 wk of age. From 8 wk to 29 wk, half of the cohort was fed ad libitum HFD and the rest continued to be fed a CD (Figure 1A ). CD composition: 18% kcal fat, 24% kcal protein, and 58% kcal of carbohydrates (Teklad Global 18% Protein Rodent Diet 2018 chow diet, Envigo, Indianapolis, USA). HFD composition: 60.3% kcal fat, 18.4% kcal protein, and 27.3% kcal of carbohydrates (Teklad Custom Diet TD.06414, Envigo). All mice were fasted overnight (from 6 pm to 9 am) prior to euthanasia. All procedures were approved by the veterinary office of canton Vaud under animal experimentation license number VD2257. In this work, proximal colons were extracted from the bio-banked samples and we did not use any new animals.
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