title: Simultaneous Two-photon in Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex.
abstract: This video shows the craniotomy procedure that allows chronic imaging of neurons in the mouse retrosplenial cortex (RSC) using in vivo two-photon microscopy in Thy1-GFP transgenic mouse line. This approach creates a possibility to investigate the correlation of behavioural manipulations with changes in neuronal morphology in vivo. The cranial window implantation procedure was considered to be limited only to the easily accessible cortex regions such as the barrel field. Our approach allows visualization of neurons in the highly vascularized RSC. RSC is an important element of the brain circuit responsible for spatial memory, previously deemed to be problematic for in vivo two-photon imaging. The cranial window implantation over the RSC is combined with an injection of mCherry-expressing recombinant adeno-associated virus (rAAV(mCherry)) into the dorsal hippocampus. The expressed mCherry spreads out to axonal projections from the hippocampus to RSC, enabling the visualization of changes in both presynaptic axonal boutons and postsynaptic dendritic spines in the cortex. This technique allows long-term monitoring of experience-dependent structural plasticity in RSC.
authors.affiliation: Department of Molecular and Cellular Neuroscience, Nencki Institute of Experimental Biology.
authors.forename: Kacper
authors.initials: K
authors.lastname: Łukasiewicz
authors.affiliation: Department of Molecular and Cellular Neuroscience, Nencki Institute of Experimental Biology.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Robacha
authors.affiliation: Department of Biochemistry, Nencki Institute of Experimental Biology.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Bożycki
authors.affiliation: Department of Molecular and Cellular Neuroscience, Nencki Institute of Experimental Biology; k.radwanska@nencki.gov.pl.
authors.forename: Kasia
authors.initials: K
authors.lastname: Radwanska
authors.affiliation: Neurobiology Centre, Nencki Institute of Experimental Biology; r.czajkowski@nencki.gov.pl.
authors.forename: Rafał
authors.initials: R
authors.lastname: Czajkowski
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology*; D003399:Craniotomy; D049229:Dendritic Spines / Q000502:physiology; D000229:Dependovirus / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006624:Hippocampus / Q000166:cytology* / Q000502:physiology; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton; D009473:Neuronal Plasticity; D017729:Presynaptic Terminals / Q000502:physiology*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein