title: Selective injection system into hippocampus CA1 via monitored theta oscillation.
abstract: 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.
authors.affiliation: Department of Neuroscience, University of Miyazaki, Faculty of Medicine, Miyazaki, Japan.
authors.forename: Jyoji
authors.initials: J
authors.lastname: Tsutajima
authors.affiliation: Department of Neuroscience, University of Miyazaki, Faculty of Medicine, Miyazaki, Japan.
authors.forename: Takato
authors.initials: T
authors.lastname: Kunitake
authors.affiliation: Department of Neuroscience, University of Miyazaki, Faculty of Medicine, Miyazaki, Japan.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Wakazono
authors.affiliation: Department of Neuroscience, University of Miyazaki, Faculty of Medicine, Miyazaki, Japan.
authors.forename: Kogo
authors.initials: K
authors.lastname: Takamiya
mesh_terms: D000818:Animals; D056547:CA1 Region, Hippocampal / Q000378:metabolism; D004396:Coloring Agents / Q000008:administration & dosage / Q000494:pharmacology; D004594:Electrophysiology; D018014:Gene Transfer Techniques / Q000295:instrumentation; D057809:HEK293 Cells; D006801:Humans; D007276:Injections, Intraventricular / Q000295:instrumentation / Q000379:methods; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008845:Microinjections / Q000295:instrumentation* / Q000379:methods*; D008863:Microspheres; D018091:Receptors, AMPA / Q000235:genetics; D013826:Theta Rhythm / Q000235:genetics
chemical_list: D004396:Coloring Agents; D018091:Receptors, AMPA