Intracerebroventricular viral injection of the neonatal mouse brain for persistent and widespread neuronal transduction

J Vis Exp. 2014 Sep 15;(91):51863. doi: 10.3791/51863.

Abstract

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene expression in the mouse brain. Here we describe a technique first introduced by Passini and Wolfe for direct intracranial delivery of virally-encoded transgenes into the neonatal mouse brain. In its most basic form, the procedure requires only an ice bucket and a microliter syringe. However, the protocol can also be adapted for use with stereotaxic frames to improve consistency for researchers new to the technique. The method relies on the ability of adeno-associated virus (AAV) to move freely from the cerebral ventricles into the brain parenchyma while the ependymal lining is still immature during the first 12-24 hr after birth. Intraventricular injection of AAV at this age results in widespread transduction of neurons throughout the brain. Expression begins within days of injection and persists for the lifetime of the animal. Viral titer can be adjusted to control the density of transduced neurons, while co-expression of a fluorescent protein provides a vital label of transduced cells. With the rising availability of viral core facilities to provide both off-the-shelf, pre-packaged reagents and custom viral preparation, this approach offers a timely method for manipulating gene expression in the mouse brain that is fast, easy, and far less expensive than traditional germline engineering.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Animals, Newborn
  • Brain / cytology
  • Brain / physiology*
  • Brain / virology*
  • Dependovirus / genetics
  • Dependovirus / physiology*
  • Female
  • Gene Transfer Techniques*
  • Genetic Vectors
  • Injections, Intraventricular
  • Male
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Neurons / physiology*
  • Neurons / virology
  • Pregnancy
  • Transduction, Genetic
  • Transgenes