title: Neurons can be labeled with unique hues by helper virus-free HSV-1 vectors expressing Brainbow.
abstract: A central problem in neuroscience is elucidating synaptic connections, the connectome. Because mammalian forebrains contain many neurons, labeling specific neurons with unique tags is desirable. A novel technology, Brainbow, creates hundreds of hues by combinatorial expression of multiple fluorescent proteins (FPs).
  
  
  
  We labeled small numbers of neurons, and their axons, with unique hues, by expressing Brainbow from a helper virus-free Herpes Simplex Virus (HSV-1) vector.
  
  
  
  The vector expresses a Brainbow cassette containing four FPs from a glutamatergic-specific promoter. Packaging HSV-Brainbow produced arrays of seven to eight Brainbow cassettes, and using Cre, each FP gene was in a position to be expressed, in different cassettes. Delivery into rat postrhinal (POR) cortex or hippocampus labeled small numbers of neurons with different, often unique, hues. An area innervated by POR cortex, perirhinal (PER) cortex, contained axons with different hues. Specific axons in PER cortex were matched to specific cell bodies in POR cortex, using hue.
  
  
  
  HSV-Brainbow is the only technology for labeling small numbers of neurons with unique hues. In Brainbow mice, many neurons contain the same hue. Brainbow-adeno-associated virus vectors require transduction of the same neuron with multiple vector particles, confounding neuroanatomical studies. Replication-competent Brainbow-pseudorabies virus vectors label multiple neurons with the same hue.
  
  
  
  Attractive properties of HSV-Brainbow include each vector particle contains multiple cassettes, representing numerous hues, recombination products are stabile, and experimental control of the number of labeled neurons. Labeling neurons with unique hues will benefit mapping forebrain circuits.
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA.
authors.forename: Guo-Rong
authors.initials: GR
authors.lastname: Zhang
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhao
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA.
authors.forename: P M Abdul
authors.initials: PMA
authors.lastname: Muneer
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Cao
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA.
authors.forename: Xu
authors.initials: X
authors.lastname: Li
authors.affiliation: New Jersey Neuroscience Institute, JFK Medical Center, Edison, NJ 08818, USA. Electronic address: alfredgeller1@gmail.com.
authors.forename: Alfred I
authors.initials: AI
authors.lastname: Geller
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D006224:Cricetinae; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism*; D008297:Male; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D017207:Rats, Sprague-Dawley; D011995:Recombination, Genetic; D050598:Vesicular Glutamate Transport Protein 1 / Q000235:genetics / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; C493466:Slc17a7 protein, rat; D050598:Vesicular Glutamate Transport Protein 1
keywords: Axon projection; Brainbow; Cre-mediated recombination; Fluorescent protein; Herpes simplex virus vector; Hue