title: Generation of transgenic marmosets expressing genetically encoded calcium indicators.
abstract: Chronic monitoring of neuronal activity in the living brain with optical imaging techniques became feasible owing to the continued development of genetically encoded calcium indicators (GECIs). Here we report for the first time the successful generation of transgenic marmosets (Callithrix jacchus), an important nonhuman primate model in neurophysiological research, which were engineered to express the green fluorescent protein (GFP)-based family of GECIs, GCaMP, under control of either the CMV or the hSyn promoter. High titer lentiviral vectors were produced, and injected into embryos collected from donor females. The infected embryos were then transferred to recipient females. Eight transgenic animals were born and shown to have stable and functional GCaMP expression in several different tissues. Germline transmission of the transgene was confirmed in embryos generated from two of the founder transgenic marmosets that reached sexual maturity. These embryos were implanted into six recipient females, three of which became pregnant and are in advanced stages of gestation. We believe these transgenic marmosets will be invaluable non-human primate models in neuroscience, allowing chronic in vivo monitoring of neural activity with functional confocal and multi-photon optical microscopy imaging of intracellular calcium dynamics.
authors.affiliation: Cerebral Microcirculation Section, Laboratory of Functional and Molecular Imaging, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jung Eun
authors.initials: JE
authors.lastname: Park
authors.affiliation: Cerebral Microcirculation Section, Laboratory of Functional and Molecular Imaging, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Xian Feng
authors.initials: XF
authors.lastname: Zhang
authors.affiliation: Cerebral Microcirculation Section, Laboratory of Functional and Molecular Imaging, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Choi
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Tonomachi, Kawasaki, Kanagawa 210-0821, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Okahara
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Tonomachi, Kawasaki, Kanagawa 210-0821, Japan.|Keio advanced Research Center, Keio University, Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
authors.forename: Erika
authors.initials: E
authors.lastname: Sasaki
authors.affiliation: Cerebral Microcirculation Section, Laboratory of Functional and Molecular Imaging, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Afonso C
authors.initials: AC
authors.lastname: Silva
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002118:Calcium / Q000378:metabolism*; D002144:Callithrix / Q000235:genetics*; D003587:Cytomegalovirus / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics*; D023421:Models, Animal; D064926:Neurophysiological Monitoring / Q000379:methods; D061848:Optical Imaging / Q000379:methods; D011401:Promoter Regions, Genetic
chemical_list: D049452:Green Fluorescent Proteins; D002118:Calcium