Treatments of mCRP and 8C10 were administered in the CA1 region of the mouse hippocampus by stereotactic surgery procedures, as previously performed [30 (link)]. Three-month-old C57BL/6J mice were anesthetized with 100 mg/kg ketamine (Ketolar 50 mg/mL, Pfizer, Alcobendas, Madrid, Spain) and 10 mg/kg xylazine (Rompun 2%, Bayer, Leverkusen, Germany) mixture i.p., and immobilized in a stereotactic apparatus (David Kopf Instruments, Tujunga, CA, USA). The experimental agent solutions were infused bilaterally into the CA1 area of the hippocampus. Injections were performed at a rate of 5 × 10−4 mL/min at coordinates relative to Bregma of −2 mm A/P, ±1.3 mm M/L, −1.6 mm V/D. Two microliters of each solution were delivered to the application point with a 2 µL 25-gauge 7000 series Neuros Syringe (Hamilton Central Europe S.R.L., Giarmata, Romania). The syringe was attached to a micro-infusion pump (Bioanalytical systems Inc., West Lafayette, IN, USA) and left in position for 5 min after delivery in order to prevent the solution from surging back.
Ketolar 50 mg ml
Manufactured by Pfizer
Sourced in Spain
Ketolar 50 mg/mL is a sterile injectable solution. It contains the active ingredient ketamine hydrochloride, which is a dissociative anesthetic agent.
Automatically generated - may contain errors
2 protocols using ketolar 50 mg ml
1
Stereotactic Injection of mCRP and 8C10 in Mouse Hippocampus
2
Astrocyte and Microglia Immunohistochemistry in Mouse Hippocampus
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The brains of three mice per experimental group were histologically processed to confirm the presence of reactive astrocytes and/or microglia in the hippocampus. Mice were perfused with 4% paraformaldehyde in phosphate buffer under anesthesia of 10 mg/kg xylazine (Rompun 2%, Bayer, Leverkusen, Germany) intraperitoneally (i.p.) and 80 mg/kg ketamine (Ketolar 50 mg/mL, Pfizer, Alcobendas, Madrid, Spain) i.p. Brains were postfixed for 24 h, cryopreserved in 30% sucrose solution, and frozen on dry ice. Twenty-µm slices were double-stained with primary antibodies against GFAP (1:1000; Dako, Glostrup, Denmark)
and Iba1 (1:500; Wako, Richmond, VA, USA). The secondary antibodies utilized included Alexa Fluor 488 and Alexa Fluor 546 (1:1,000; Molecular Probes, Thermo Fisher Scientific).
Tissue sections were mounted on Mowiol ® (Calbiochem, San Diego, CA, USA).
and Iba1 (1:500; Wako, Richmond, VA, USA). The secondary antibodies utilized included Alexa Fluor 488 and Alexa Fluor 546 (1:1,000; Molecular Probes, Thermo Fisher Scientific).
Tissue sections were mounted on Mowiol ® (Calbiochem, San Diego, CA, USA).
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