All procedures were approved by Johns Hopkins University Animal Care and Use Committee. Male and female transgenic mice (ChAT-cre, ChAT-cre/jRGECO1a) between 6 and 16 weeks were used for the experiments. All experiments (passive recording and chemogenetic suppression) used ChAT-cre mice unless stated otherwise (Supplementary Table 1 ). ChAT-cre mice were obtained from the Jackson Laboratory (stock no. 006410) and bred in-house. ChAT-cre/jRGECO1a mice were bred in-house by crossing homozygous female ChAT-cre mice and hemizygous male jRGECO1a mice obtained from the Jackson Laboratory (stock no. 030526). Offspring genotypes were confirmed by PCR (Lucigen EconoTaq Plus GREEN 2X). Both heterozygous and homozygous ChAT-cre/jRGECO1a mice were used in the experiments and no phenotypic difference were observed.
Chat cre mice
Manufactured by Jackson ImmunoResearch
Sourced in Montenegro, China, France
ChAT-Cre mice are a genetically modified mouse model where the Cre recombinase enzyme is expressed under the control of the choline acetyltransferase (ChAT) promoter. The ChAT enzyme is responsible for the synthesis of acetylcholine, a neurotransmitter. This model allows for the selective manipulation of cholinergic neurons in the mouse.
Automatically generated - may contain errors
Lab products found in correlation
C57bl 6j mice, by Janvier Labs (2 mentions)
Chat cre, by Jackson ImmunoResearch (2 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (2 mentions)
Hsod1g93a mice, by Jackson ImmunoResearch (1 mentions)
R26 idtr mice, by Jackson ImmunoResearch (1 mentions)
Vgat cre mice, by Jackson ImmunoResearch (1 mentions)
Vglut2 cre mice, by Jackson ImmunoResearch (1 mentions)
Chatbac egfp mice, by Jackson ImmunoResearch (1 mentions)
Cdk5f f mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6 animals, by Jackson ImmunoResearch (1 mentions)
B6.129x1 thtm1 cre te kieg, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Jackson ImmunoResearch (1 mentions)
16 protocols using chat cre mice
1
Transgenic Mouse Experiments in Cholinergic Neurobiology
2
Social Defeat Stress in ChAT-Cre Mice
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Adult female C57BL/6 J mice (8 weeks; Janvier Labs, France) were used as experimental subjects. Mice were group housed (5 per cage), and habituated to housing conditions and experimenter handling for 1 week before starting the social defeat stress procedure. Adult male CD1 mice (former breeder aged > 4 months, Janvier Labs, France) were used as aggressors. CD1 mice were housed alone and screened during 3 days to assess their aggressive behavior towards female mice (latency to attack less than 1 min). Heterozygous female choline acetyltransferase ChAT-IRES-Cre Knock-In mice (ChAT-Cre mice, The Jackson Laboratory, stock number: 006410) were backcrossed for 9 generations on a C57BL/6 J and used to specifically target cholinergic neurons. All mice were maintained in a 12 h–12 h light-dark cycle at a room temperature of 21–22 °C, with food and water available ad libitum. All behavioral tests took place during the light cycle, with exception of the sucrose preference test (see below). All procedures were in accordance with the recommendations of the European Commission (2010/63/EU) for care and use of laboratory animals, and approved by the French National Ethical Committee. The number of mice used for each behavioral and electrophysiological experiments can be found in the Supplementary Table.
3
Generating Triple-Transgenic ALS Mouse Model
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In order to get triple-transgenic hSOD1G93A/ChATCre/Srfloxp/loxp animals, ChATCre mice (The Jackson Laboratory, strain no. 006410) were first crossed with Srf-floxed mice (52 (link), 73 (link)). It is worth noting that ChATCre mice allowed for the deletion of cholinergic cells such as MNs as well as other cell types, including skin cells (74 (link)). This mouse double-transgenic line (ChATCre/Srfloxp/loxp) was further crossed to high-copy hSOD1G93A mice (The Jackson Laboratory, strain no. 002298) to get hSOD1G93A/ChATCre/Srfloxp/loxp triple-transgenic mice. This allowed for the motor neuron–specific Srf KO in an ALS mouse model. This triple-transgenic mouse line was maintained by mating hSOD1G93A/ChATCre/Srfloxp/wt males to hSOD1–/ChATCre/Srfloxp/wt females. From these breedings, 4 groups were derived: (a) WT (hSOD1–/ChATCre/Srfwt/wt), (b) mSOD1 (hSOD1G93A/ChATCre/Srfwt/wt); (c) Srf KO (hSOD1–/ChATCre/Srfloxp/loxp), and (d) mSOD1/Srf KO (hSOD1G93A/ChATCre/Srfloxp/loxp).
Mice were kept with free access to food and water in a pathogen-free animal facility at the University of Ulm, with a 12-hour day-night shift and appropriate temperature and humidity conditions. Humane endpoints of mSOD1 and mSOD1/Srf-KO mice were determined individually according to the criteria indicated below.
Mice were kept with free access to food and water in a pathogen-free animal facility at the University of Ulm, with a 12-hour day-night shift and appropriate temperature and humidity conditions. Humane endpoints of mSOD1 and mSOD1/Srf-KO mice were determined individually according to the criteria indicated below.
4
Genetic Manipulation of Cholinergic Neurons
5
Genetic Manipulation of Cholinergic Neurons
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All animal care and experimental procedures were approved in advance by the University of Virginia Institutional Animal Care and Use Committee. The sNuc-Seq experiments used adult male Chat-Cre mice (Jackson Laboratories, JAX, stock # 28861; 28 weeks old). The fluorescent in situ hybridization experiments used C57BL/6J mice from the Jackson Laboratory (JAX, 000664). The following mouse lines were used for optogenetic studies: Chat-Cre (Rossi et al., 2011 (link); JAX, 028861); Crfr2α-eGFPCre bacterial artificial chromosome (BAC) transgenic mice (“Crhr2-Cre mice”; Anthony et al., 2014 (link)); Vipr2-Cre (Daigle et al., 2018 (link); JAX, 31332); Phox2b-Flp (Hirsch et al., 2013 (link); JAX, 22407); CaTCh (Daigle et al., 2018 (link); JAX, 025109); Ai32 (RCL-ChR2(H134R)/EYFP) mice (Madisen et al., 2012 (link); JAX, 024109); and a Chat-p2a-Flp mouse line (see Method Details below). The anterograde tracing experiments used Chat-Cre, Crhr2-Cre, Vipr2-Cre, and C57BL/6J mice. Unless otherwise specified, all experiments used adult mice with approximately equal numbers of male and female mice. Mice were housed at 22-24°C with a 12-h light:12-h dark cycle and ad libitum access to standard mouse chow and water.
6
Conditional Genetic Knockout Experiments
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All animals in this study were male and their age was 8–20 weeks. Chrm4-floxed and D1RCre lines are described in the literature (Jeon et al., 2010 (link)) and ChATCre mice were purchased from The Jackson Laboratory. Mice were on a C57BL/6N background with minor (<15%) contributions from 129SvEv and C57BL/6J. Wildtype littermate animals carrying two floxed alleles of the M4R were used as controls. The control mice did not display significant differences in baseline (BL) behaviors in the place preference and operant runway tests and were therefore pooled to one control group in these experiments. Mice were single-housed 48 h prior to experiments, housed with environmental enrichment and kept in a pathogen-free facility on a regular 12-h light/dark cycle. All experiments were performed during the light phase. Food and water were provided ad libitum with the exception of animals in the 5CSRTT and the food-reward operant runway (as detailed in the respective “Materials and Methods” section). The use of animals for this study followed the EU directive 2010/63/EU for animal experiments and were approved by the Research Animal Care and Use Committee in Linköping, Sweden.
7
Murine Behavioral Experiments Using Cre-Expressing Mice
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The Institutional Animal Care and Use Committee of the Beijing Institute of Technology in Beijing, China approved all surgical and experimental procedures. The following animals were used in this study: adult (2–4 months old) male wild-type (WT) mice (C57/BL6) and ChAT-cre mice (Jackson laboratory Stock no. 006140). Mice were group-housed in a 12/12 h light/ dark cycle (2–5 animals per cage) at a consistent ambient temperature of 23 ± 1 °C and humidity of 50 ± 5%. All experiments were performed during the light cycle. Food and water were accessed ad libitum. Littermates were randomly assigned to each condition by the experimenter.
8
Genetically Engineered Mouse Lines
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All animal protocols were approved by the Institutional Animal Care and Use Committee at Massachusetts General Hospital (protocols #2009N000239 and #2013N000115). All methods were performed in accordance with relevant regulations. The following mouse lines were obtained from Jackson Laboratory (Bar Harbor, ME): ChAT::Cre mice (stock #006410) were crossed with R26-iDTR mice (stock #007900) to obtain ChAT::Cre;R26-iDTR (annotated as ChAT-iDTR) mice. ChAT::Cre mice also were crossed with R26-ChR2-tdTomato (stock #012567) reporter mice to obtain ChAT::Cre;R26-ChR2-tdTomato (annotated as ChAT-ChR2) mice, in which ChAT+ cells express the light-sensitive ion channel ChR2 and red fluorescent protein, tdT. Cx3cr1GFP;CCR2RFP mice (stock #032127), in which red fluorescent protein is expressed by peripheral monocytes and lymphocytes, and green fluorescent protein is expressed by macrophages. All experiments used adult mice (both sexes) between 10 and 12 weeks of age.
9
Cholinergic Neuron Expression in Mice
10
Choline Acetyltransferase-Cre Mice Study
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All procedures were approved by the Animal Care and Use Committee (Wuhan Institute of Physics and Mathematics, the Chinese Academy of Sciences). Experiments were performed on adult male C57BL/6 mice purchased from Hunan SJA Laboratory Animal (Changsha, Hunan, China), and ChAT-Cre mice with C57BL/6 background were from Jackson Laboratory (Bar Harbor, ME, USA). The animals were housed on a 12/12 light/dark cycle with ambient temperature (21 ± 1°C) and humidity (50 ± 5%). The adult ChAT-Cre mice of both sexes (2–3 months old, 20–30 g) and the male C57BL/6 mice (8 weeks old, 20–30 g) were used in the study.
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