title: Long-Term Two-Photon Calcium Imaging of Neuronal Populations with Subcellular Resolution in Adult Non-human Primates.
abstract: Two-photon imaging with genetically encoded calcium indicators (GECIs) enables long-term observation of neuronal activity in vivo. However, there are very few studies of GECIs in primates. Here, we report a method for long-term imaging of a GECI, GCaMP6f, expressed from adeno-associated virus vectors in cortical neurons of the adult common marmoset (Callithrix jacchus), a small New World primate. We used a tetracycline-inducible expression system to robustly amplify neuronal GCaMP6f expression and up- and downregulate it for more than 100 days. We succeeded in monitoring spontaneous activity not only from hundreds of neurons three-dimensionally distributed in layers 2 and 3 but also from single dendrites and axons in layer 1. Furthermore, we detected selective activities from somata, dendrites, and axons in the somatosensory cortex responding to specific tactile stimuli. Our results provide a way to investigate the organization and plasticity of cortical microcircuits at subcellular resolution in non-human primates.
authors.affiliation: Division of Brain Biology, National Institute for Basic Biology, Aichi 444-8585, Japan; Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Aichi 444-8585, Japan; Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Brain Science Institute, Saitama 351-0198, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Sadakane
authors.affiliation: Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Aichi 444-8585, Japan; Division of Brain Circuits, National Institute for Basic Biology, Aichi 444-8585, Japan.
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Masamizu
authors.affiliation: Division of Brain Biology, National Institute for Basic Biology, Aichi 444-8585, Japan; Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Aichi 444-8585, Japan; Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Brain Science Institute, Saitama 351-0198, Japan.
authors.forename: Akiya
authors.initials: A
authors.lastname: Watakabe
authors.affiliation: Division of Brain Circuits, National Institute for Basic Biology, Aichi 444-8585, Japan; Laboratory of Cell Recognition and Pattern Formation, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Terada
authors.affiliation: Division of Brain Biology, National Institute for Basic Biology, Aichi 444-8585, Japan; Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Brain Science Institute, Saitama 351-0198, Japan.
authors.forename: Masanari
authors.initials: M
authors.lastname: Ohtsuka
authors.affiliation: Division of Brain Biology, National Institute for Basic Biology, Aichi 444-8585, Japan; Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Brain Science Institute, Saitama 351-0198, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Takaji
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mizukami
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Keiya
authors.initials: K
authors.lastname: Ozawa
authors.affiliation: Department of Medical Neuroscience, Graduate School of Medicine, Kanazawa University, Ishikawa 920-8640, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawasaki
authors.affiliation: Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Aichi 444-8585, Japan; Division of Brain Circuits, National Institute for Basic Biology, Aichi 444-8585, Japan. Electronic address: mzakim@nibb.ac.jp.
authors.forename: Masanori
authors.initials: M
authors.lastname: Matsuzaki
authors.affiliation: Division of Brain Biology, National Institute for Basic Biology, Aichi 444-8585, Japan; Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Aichi 444-8585, Japan; Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Brain Science Institute, Saitama 351-0198, Japan. Electronic address: tetsuo.yamamori@riken.jp.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Yamamori
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002144:Callithrix; D003712:Dendrites / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D004317:Doxorubicin / Q000633:toxicity; D052139:Intracellular Calcium-Sensing Proteins / Q000235:genetics / Q000378:metabolism; D036641:Microscopy, Fluorescence, Multiphoton; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D013003:Somatosensory Cortex / Q000187:drug effects / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology
chemical_list: D052139:Intracellular Calcium-Sensing Proteins; D004317:Doxorubicin; D013752:Tetracycline; D002118:Calcium