Here we demonstrate a technique for widespread neuronal transduction by intraventricular injection of adeno-associated virus into the neonatal mouse brain. This method provides a rapid and easy way to attain lifelong expression of virally-delivered transgenes.
This research was supported by the Robert A. and Rene E. Belfer Family Foundation, NIA R21 AG038856 (JLJ), BrightFocus Foundation Alzheimer’s Disease research grant A2010097 (JLJ), and NIA Biology of Aging Training grant T32 AG000183 (support for SDG).
Materials
Name of Material/ Equipment
Company
Catalog Number
Comments/Description
ICR outbred mice
Harlan
Hsd:ICR (CD-1)
This strain is also known as CD-1
FVB inbred mice
The Jackson Laboratory
1800
5-6 weeks of age
Nestlets
Lab Supply
NESTLETS
Shepherd shacks
Lab Supply
SS-mouse
High fat rodent chow
Purina Mills
PicoLab Mouse diet 20, #5058
This is our standard breeder chow
High fat rodent chow (alternative)
Harlan Laboratories
Teklad Global 19% protein rodent diet #2019S
If low phytoestrogen, autoclavable diet is needed
Injection syringe
Hamilton
7653-01
10 ml syringe
Injection needles
Hamilton
7803-04, RN 6PK PT4
32 gauge, for standard P0 injections
Metal plate for cryoanesthesia
McMaster Carr
8975K439
Raw aluminum plate, 6” x 12”, 0.25” thick, will need to be cut into 3 equal pieces and edges sanded by local machine shop
Small animal stereotaxic device with digital readout
David Kopf Instruments
Model 940
Universal syringe holder with needle support foot
David Kopf Instruments
Model 1772-F1
Neonatal frame
Stoelting
51625
Officially called a mouse and neonatal rat adaptor
Biohazard disposal bags with sterile indicator
VWR
14220-030
Important! – Check with local veterinary and environmental safety staff to learn your institute’s protocol for biohazard waste disposal
Kim, J., Grunke, S. D., Levites, Y., Golde, T. E., Jankowsky, J. L. Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction. J. Vis. Exp. (91), e51863, doi:10.3791/51863 (2014).