We used 406 male and female mice (7–8 weeks old). Mice were allowed to acclimate for seven days before beginning the experiments. They were housed in individually ventilated cages, up to 5 mice per cage on 12-h dark/light cycle and allowed free access to standard dry rodent diet and water, supplemented with 1g/l L- cysteine (AppliChem, Darmstadt, Germany) [11]. Analgesia was achieved with paracetamol suppositories, in order to avoid interactions with the immune system.

H2S synthesis is catalyzed by three different enzymatic systems, cystathionine γ- lyase (CSE), which is the key enzyme for H2S synthesis in the liver and kidney, cystathionine β- synthase (CBS), which is overexpressed in brain tissue and 3-mercaptopyruvate sulfurtransferase (3MST) which is ubiquitously expressed [12]. We investigated the role only of CSE and 3MST, due to the fact that homozygotic CBS deficiency is related to a neonatal mortality of over 90%. [13,14]. Cse -/- mice were created after breeding of Cse +/- mice (Friedrich Shiller University Hospital, Jena, Germany). The Cse+/- mice were generated as previously described [13]. In short chimeric male Cse +/- mice were produced after injection of Cth+/- stem cells into C57BL/6 blastocysts. These were then crossed with C57BL/6 female mice to obtain Cse +/- pups. Cse +/- pups were backcrossed multiple times to achieve high genetic homogenicity on C57BL/6 background. The Cse +/- males and females produced were finally bred to obtain wild- type (Cse +/+), Cse +/-, and Cse -/- littermates. 3Mst+/+ and 3Mst-/- mice were created, as described previously [15].

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.