Selective injection system into hippocampus CA1 via monitored theta oscillation

Methods of cell biology and electrophysiology using dissociated primary cultured neurons allow in vitro study of molecular functions; however, analysis of intact neuronal circuitry is often preferable. To investigate exogenous genes, viral vectors are most commonly injected using a pipette that is i...

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Bibliographic Details
Published in:PloS one 2013-12, Vol.8 (12), p.e83129-e83129
Main Authors: Tsutajima, Jyoji, Kunitake, Takato, Wakazono, Yoshihiko, Takamiya, Kogo
Format: Article
Language:English
Subjects:
Analysis
Animals
Biomonitoring
Brain
CA1 Region, Hippocampal - metabolism
Circuits
Coloring Agents - administration & dosage
Coloring Agents - pharmacology
Copper
Electrodes
Electrophysiology
Expression vectors
Gene expression
Gene therapy
Gene transfer
Gene Transfer Techniques - instrumentation
Genetic vectors
HEK293 Cells
Hippocampus
Humans
Injection
Injections, Intraventricular - instrumentation
Injections, Intraventricular - methods
Male
Medical research
Medicine
Memory
Mice
Mice, Inbred C57BL
Mice, Knockout
Microinjections - instrumentation
Microinjections - methods
Microspheres
Molecular modelling
Neural networks
Neurons
Neurophysiology
Neurosciences
Personal computers
Polyethylene
Receptors, AMPA - genetics
Researchers
Rodents
Science
Studies
Theta Rhythm - genetics
Vectors (Biology)
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Description
Summary:Methods of cell biology and electrophysiology using dissociated primary cultured neurons allow in vitro study of molecular functions; however, analysis of intact neuronal circuitry is often preferable. To investigate exogenous genes, viral vectors are most commonly injected using a pipette that is inserted from the top of the cortex. Although there are few reports that describe the success rate of injection in detail, it is sometimes difficult to locate the pipette tip accurately within the CA1 pyramidal cell layer because the pyramidal layer is only 0.1 mm thick. In the present study, we have developed a system to inject viral vectors accurately into the mouse hippocampal CA1 pyramidal cell layer using a stereotaxic injection system with simultaneous electrophysiological monitoring of theta oscillation. The pipette tip was positioned reliably based on integrated values of the theta oscillation in the hippocampal CA1 pyramidal cell layer. This approach allows accurate injection of solutions and provides an efficient method of gene transfer using viral vectors into the hippocampus, which can be a useful tool for studies involving the molecular mechanisms of neuronal functions.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0083129