title: Molecular profiling of neurons based on connectivity.
abstract: The complexity and cellular heterogeneity of neural circuitry presents a major challenge to understanding the role of discrete neural populations in controlling behavior. While neuroanatomical methods enable high-resolution mapping of neural circuitry, these approaches do not allow systematic molecular profiling of neurons based on their connectivity. Here, we report the development of an approach for molecularly profiling projective neurons. We show that ribosomes can be tagged with a camelid nanobody raised against GFP and that this system can be engineered to selectively capture translating mRNAs from neurons retrogradely labeled with GFP. Using this system, we profiled neurons projecting to the nucleus accumbens. We then used an AAV to selectively profile midbrain dopamine neurons projecting to the nucleus accumbens. By comparing the captured mRNAs from each experiment, we identified a number of markers specific to VTA dopaminergic projection neurons. The current method provides a means for profiling neurons based on their projections.
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Mats I
authors.initials: MI
authors.lastname: Ekstrand
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Nectow
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Zachary A
authors.initials: ZA
authors.lastname: Knight
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Kaamashri N
authors.initials: KN
authors.lastname: Latcha
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Pomeranz
authors.affiliation: Laboratory of Molecular Genetics, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA. Electronic address: friedj@rockefeller.edu.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Friedman
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000378:metabolism; D047468:Immunoprecipitation; D008822:Mice, Transgenic; D009446:Neurobiology / Q000379:methods*; D059906:Neuroimaging / Q000379:methods*; D009474:Neurons / Q000166:cytology*; D009714:Nucleus Accumbens / Q000166:cytology; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000737:chemistry*
chemical_list: D000906:Antibodies; D049452:Green Fluorescent Proteins