For in vivo experiments, Stab1 KO (B6.129S2-Stab1tm1.1Cger 16) [12 (link)] were used. Homozygous littermates or purchased C57Bl/6J wildtype (Janvier Labs, Le Genest-Saint-Isle, France) served as Ctrl. Heterozygous mice were not used. All animals were hosted in single ventilated cages (Sealsafe plus DGM™, Techniplast, Buguggiate, Italy; Bedding H0234-20, Ssniff, Soest, Germany) in a 12 h/12 h day/night cycle under specific-pathogen-free conditions and fed ad libitum with a standard rodent diet (ssniff®®R/M-H autoclavable, V1534-000, Ssniff, Soest, Germany). For DNA extraction and genotyping, the KAPA HotStart Mouse Genotyping Kit (KK7352, Merck, Darmstadt, Germany) and primers (Metabion international AG, Planegg/Steinkirchen, Germany) were utilized (Supplementary Table S1 ).
C57bl 6j wild type
Manufactured by Janvier Labs
Sourced in France
The C57BL/6J wild-type is a commonly used laboratory mouse strain. It serves as a standard reference strain for various research applications.
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4 protocols using c57bl 6j wild type
1
In vivo Stab1 KO Mouse Protocol
2
Investigating Endocannabinoid Regulation in Mice
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Experimental procedures followed French and European guidelines for animal experimentation and in compliance with the institutional animal welfare guidelines of the Paris Brain Institute. Experiments for paired recordings were performed on both sexes aged between P30 and P40 CB1-tdTomato mice (Winters et al., 2012 (link)), except for experiments illustrated in Figure S6 , for which mice were older than P60. In some experiments, CB1-tdTomato mice were crossed with GAD67-GFP mice to identify CB1-expressing INs for cell counting. In experiments for Figures 1 and S6 , C57BL/6J wild-type were purchased from Janvier laboratories. Mice were housed in an animal facility with a 12h light/dark cycle, with food and water available ad libitum. Viral injections and chronic cranial windows for 2P Ca2+ imaging experiments in vivo were performed on two months old male mice of CB1-tdTomato or CB1floxed/floxed mice. Imaging sessions started following habituation, four to five weeks post-surgery for CB1-tdTomato or CB1floxed/floxed mice, respectively.
3
In Utero Electroporation of Mouse Embryos
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C57BL/6J wild-type (Janvier) mice were used, except for Fig. S1F , for which the Btg2::GFP line was used (Haubensak et al., 2004 (link)). The morning of the vaginal plug was defined as E0 and 13 days later (E13) mice were deeply anesthetized with isoflurane, their uterine horns exposed and ∼2 µl of plasmid (2 µg/µl with 0.01% of Fast Green) injected into the lateral ventricle of the embryonic brains. Electrodes were placed around the embryo head, with the anode facing the injection site, and six pulses of 30 V for 5 ms each were delivered using an electroporator (BTX ECM830). Afterwards, the uterus was relocated within the abdominal cavity, and the muscular walls and overlying skin were sutured independently. Mice were transferred to the housing box when fully awake and sacrificed at the indicated time points by cervical dislocation. Eventually, mice were administered with a single intraperitoneal dose of EdU (1 mg/kg) 24 h before sacrifice. All animal procedures were approved by local authorities and complied with all relevant ethical regulations (TVV 16/2018).
4
Standardized Mice Husbandry and Experiments
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C57BL/6J wild‐type were purchased from JANVIER LABS (Saint‐Berthevin Cedex, France). They were housed in a 12‐hour light‐dark cycle in the animal facility of the University of Lübeck (Lübeck, Germany) and fed with standard chow diet. All experiments were performed in 8‐ to 12‐week‐old age‐ and sex‐matched mice by certified personnel. All experiments had been permitted by the state government of Schleswig‐Holstein.
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C57BL/6J wild‐type were purchased from JANVIER LABS (Saint‐Berthevin Cedex, France). They were housed in a 12‐hour light‐dark cycle in the animal facility of the University of Lübeck (Lübeck, Germany) and fed with standard chow diet. All experiments were performed in 8‐ to 12‐week‐old age‐ and sex‐matched mice by certified personnel. All experiments had been permitted by the state government of Schleswig‐Holstein.
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