Mar 14, 2024

Public workspaceStereotaxic α-Syn PFF injection into mouse striatum

Forked from a private protocol
DOI
dx.doi.org/10.17504/protocols.io.dm6gp34p1vzp/v1
  • Ramhari Kumbhar1,2,3,
  • Hanseok Ko1,2,3,
  • Valina L. Dawson4,5,3,6,7,8,
  • Ted Dawson4,5,3,6,7,8,9,
  • Xiaobo Mao4,5,3,6,7,8,9
  • 1Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA;
  • 2Department of Neurology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA;
  • 3Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA;
  • 4Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 5Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 6Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA 70130-2685, USA;
  • 7Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205 USA;
  • 8Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 9Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA
Eileen Ruth Torres
Open access
DOI: dx.doi.org/10.17504/protocols.io.dm6gp34p1vzp/v1
Protocol Citation: Ramhari Kumbhar, Hanseok Ko, Valina L. Dawson, Ted Dawson, Xiaobo Mao 2024. Stereotaxic α-Syn PFF injection into mouse striatum. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp34p1vzp/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 14, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 96713
Keywords: ASAPCRN
Funders Acknowledgement:
Aligning Science for Parkinson's
Grant ID: 020608
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.
Materials
  • Anesthetic- isoflurane OR pentobarbital (45 mg/kg) diluted 1:1 with 0.9% NaCl OR ketamine cocktail (ketamine 100mg/kg; xylazine 20mg/kg; acepromazine 3mg/kg i.p. or s.c.)
  • Small electric shaver
  • scalpel
  • 26 gauge needle and Hamilton syringe
  • minisyringe pump (Model BS-800, Braintree Scientific)
  • sterile wound healing clips (7 mm stainless steel wound clip; Reflex 7 skin closure system) or sutures
  • bupivacaine (0.4-0.8 ml/kg of a 0.25% solution) OR 0.5% xylocaine 1.4 ml/kg, injected sc, injected) OR 0.5% xylocaine
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.
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.
Protocol references
Kwon SH, Kim S, Park AY, Lee S, Gadhe CG, Seo BA, Park JS, Jo S, Oh Y, Kweon SH, Ma SX, Kim WR, Kim M, Kim H, Kim JE, Lee S, Lee J, Ko HS. A Novel, Selective c-Abl Inhibitor, Compound 5, Prevents Neurodegeneration in Parkinson's Disease. J Med Chem. 2021 Oct 28;64(20):15091-15110. doi: 10.1021/acs.jmedchem.1c01022. Epub 2021 Sep 28. Erratum in: J Med Chem. 2021 Dec 23;64(24):18237. PMID: 34583507.