Injection of adeno-associated virus containing optogenetic and chemogenetic probes into the neonatal mouse brain

Nhan C. Huynh, Baher A. Ibrahim, Christopher M. Lee, Mickeal N. Key, Daniel A. Llano

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Optogenetics and chemogenetics are neuromodulation techniques used to study neuronal pathways. Viral vectors containing optogenetic or chemogenetic probes require at least a week after injection to produce effective expression levels in neurons. Therefore, injections need to be done in neonatal mice to enable experiments that require mice younger than 30 days of age. Here, we describe a protocol for performing surgery on neonatal mice and brain injection of a viral vector. The procedure utilizes cryoanesthesia and a pressure injector with a micropipette that can be directly injected through the skull of neonatal mice. Compared to other approaches, this protocol is relatively easy to implement and takes only minutes to perform, additionally allowing for increased numbers of injections. In the examples shown in this chapter, viral vectors were successfully delivered into the auditory cortex and the hippocampus as indicated by labeled expression of the optogenetic/chemogenetic probes. This technique provides a method for investigators to perform surgical injections and optogenetic/chemogenetic experiments in neonatal mice.

Original languageEnglish (US)
Title of host publicationNeuromethods
PublisherHumana Press Inc.
Pages19-43
Number of pages25
DOIs
StatePublished - Jan 1 2020

Publication series

NameNeuromethods
Volume152
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Fingerprint

Optogenetics
Dependovirus
Viruses
Brain
Injections
Cryoanesthesia
Surgery
Neurons
Experiments
Auditory Cortex
Skull
Hippocampus
Research Personnel
Pressure

Keywords

  • Adeno-associated virus
  • Chemogenetics
  • Cryoanesthesia
  • Neonatal mouse
  • Optogenetics
  • Rodent
  • Surgery

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Psychiatry and Mental health

Cite this

Huynh, N. C., Ibrahim, B. A., Lee, C. M., Key, M. N., & Llano, D. A. (2020). Injection of adeno-associated virus containing optogenetic and chemogenetic probes into the neonatal mouse brain. In Neuromethods (pp. 19-43). (Neuromethods; Vol. 152). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9944-6_2

Injection of adeno-associated virus containing optogenetic and chemogenetic probes into the neonatal mouse brain. / Huynh, Nhan C.; Ibrahim, Baher A.; Lee, Christopher M.; Key, Mickeal N.; Llano, Daniel A.

Neuromethods. Humana Press Inc., 2020. p. 19-43 (Neuromethods; Vol. 152).

Research output: Chapter in Book/Report/Conference proceedingChapter

Huynh, NC, Ibrahim, BA, Lee, CM, Key, MN & Llano, DA 2020, Injection of adeno-associated virus containing optogenetic and chemogenetic probes into the neonatal mouse brain. in Neuromethods. Neuromethods, vol. 152, Humana Press Inc., pp. 19-43. https://doi.org/10.1007/978-1-4939-9944-6_2
Huynh, Nhan C. ; Ibrahim, Baher A. ; Lee, Christopher M. ; Key, Mickeal N. ; Llano, Daniel A. / Injection of adeno-associated virus containing optogenetic and chemogenetic probes into the neonatal mouse brain. Neuromethods. Humana Press Inc., 2020. pp. 19-43 (Neuromethods).
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