High-resolution in vivo imaging of hippocampal dendrites and spines.
Structural changes in hippocampal dendrites and dendritic spines are thought to be a consequence of a wide range of experience- and activity-dependent manipulations. We explored the dynamics of hippocampal dendritic spines in vivo by developing a surgical preparation of the adult mouse brain that enabled two-photon imaging of fluorescently labeled CA1 pyramidal neurons. Dendritic trees and spines were repeatedly visualized over many hours in exquisite detail. We tested spine stability under both control conditions and during prolonged epileptic seizures. Remarkably, spines remained structurally stable after 30 min of experimental induction of epileptic seizures. Spines began to disappear only several hours after induction of epileptic activity. We thus demonstrate that this technique provides a methodology for direct in vivo optical studies of the intact mammalian hippocampus.
Mizrahi
Adi
A
Howard Hughes Medical Institute and Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710, USA. mizrahi@neuro.duke.edu
Crowley
Justin C
JC
Shtoyerman
Eran
E
Katz
Lawrence C
LC
D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D003712:Dendrites / Q000187:drug effects / Q000648:ultrastructure*; D004569:Electroencephalography; D004827:Epilepsy / Q000139:chemically induced / Q000473:pathology / Q000503:physiopathology; D018756:GABA Antagonists / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000187:drug effects / Q000503:physiopathology / Q000648:ultrastructure*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D010862:Pilocarpine; D017966:Pyramidal Cells / Q000378:metabolism / Q000648:ultrastructure*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D018756:GABA Antagonists; D008164:Luminescent Proteins; D010862:Pilocarpine; D049452:Green Fluorescent Proteins; D001640:Bicuculline