Cell-type-specific labeling of synapses in vivo through synaptic tagging with recombination.
The study of synaptic specificity and plasticity in the CNS is limited by the inability to efficiently visualize synapses in identified neurons using light microscopy. Here, we describe synaptic tagging with recombination (STaR), a method for labeling endogenous presynaptic and postsynaptic proteins in a cell-type-specific fashion. We modified genomic loci encoding synaptic proteins within bacterial artificial chromosomes such that these proteins, expressed at endogenous levels and with normal spatiotemporal patterns, were labeled in an inducible fashion in specific neurons through targeted expression of site-specific recombinases. Within the Drosophila visual system, the number and distribution of synapses correlate with electron microscopy studies. Using two different recombination systems, presynaptic and postsynaptic specializations of synaptic pairs can be colabeled. STaR also allows synapses within the CNS to be studied in live animals noninvasively. In principle, STaR can be adapted to the mammalian nervous system.
Department of Biological Chemistry, Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Yi
Y
Chen
Department of Biological Chemistry, Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Orkun
O
Akin
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147-2408, USA.
Aljoscha
A
Nern
Department of Biological Chemistry, Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
C Y Kimberly
CY
Tsui
Department of Biological Chemistry, Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Matthew Y
MY
Pecot
Department of Biological Chemistry, Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: lzipursky@mednet.ucla.edu.
S Lawrence
SL
Zipursky
D000818:Animals; D030801:Animals, Genetically Modified; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008526:Medulla Oblongata / Q000166:cytology*; D046529:Microscopy, Electron, Transmission; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000648:ultrastructure; D017729:Presynaptic Terminals / Q000378:metabolism*; D017981:Receptors, Neurotransmitter / Q000235:genetics / Q000378:metabolism; D045522:Recombinases / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D013569:Synapses / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014795:Visual Pathways / Q000166:cytology / Q000378:metabolism
C510146:BRP protein, Drosophila; D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D017981:Receptors, Neurotransmitter; D045522:Recombinases; D014157:Transcription Factors; C033652:postsynaptic density proteins