Stereotaxic α-Syn PFF injection into mouse striatum
Ted Dawson, Xiaobo Mao, Ramhari Kumbhar, Hanseok Ko, Valina L. Dawson
Abstract
For stereotaxic injection of α-Syn PFF into the mouse striatum using 2-3 month old mice. This forked protocol has slight variations in anesthesia solution and injection coordinations.
Before start
Personnel should use aseptic techniques and conduct the surgery using sterile gloves in a sterile environment. Surgical instruments should be sterilized before surgeries using an autoclave and during the procedure using glass bead
sterilizer.
Steps
Anesthetize mice with a ketamine cocktail (ketamine 100mg/kg; xylazine 10mg/kg; i.p. or s.c.).
Shave the animals’ skulls.
Create a 1.5 cm incision in the scalp after cleaning the scalp with alcohol or betadine swabs.
Retract the underlying fascia will and drill a hole in the skull.
Insert a 26 gauge needle attached to a 2uL Hamilton syringe containing recombinant α-syn PFF (5 μg/2 μL) or an equivalent volume of PBS into the striatum unilaterally (right hemisphere) at coordinates
mediolateral 2.0 mm from bregma, anteroposterior -0.2 mm, and dorsoventral 2.8mm. at a rate of 0.4 ul per minute using a minisyringe pump (Model BS-800, Braintree Scientific).
Leave the needle in place for 5 minutes before withdrawing the needle.
Close the incision using sterile wound healing clips (7 mm stainless steel wound clip; Reflex 7 skin closure system) or sutures.
Immediately after the stereotaxic procedure, treat the stapled wound site with bupivacaine (0.4-0.8 ml/kg of a 0.25% solution, injected sc) or 0.5% xylocaine 1.4 ml/kg, injected sc, injected) or 0.5% xylocaine when necessary or as determined by animal use policy.
