This proposal outlines the development of novel methods for the mapping of complex neural pathways and the specification of the neurons that compose these pathways. Our groups have recently made a recombinant Pseudorabies virus (PRV) that is conditional on cre-recombinase for viability and expression of a GFP reporter gene. This virus has been used successfully to map inputs to specific neuronal populations in defined brain regions. For simplicity we will refer to this and related methods as PAM, Pseudorabies virus assisted mapping. In this application, we propose to further develop PAM so as to allow: 1) Simultaneous Retrograde Tracing from Different Neurons in the Same Brain Region, 2) Simultaneous Retrograde Tracing from Similar Neurons in Different Brain Regions, 3) Anterograde tracing 4) Identification of Neurons Projecting to More than One Site 5) Specification of the neurons that are identified in these mapping studies. To establish the validity of the approach, these studies will be performed in analyses of hypothalamic circuits that regulate feeding. Once the methods are validated, the viruses and animals will be made freely available to other investigators interested in analogous studies of other neuronal pathways.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA018799-05
Application #
7580993
Study Section
Special Emphasis Panel (ZRG1-MDCN-B (55))
Program Officer
Wu, Da-Yu
Project Start
2005-02-15
Project End
2010-11-30
Budget Start
2008-12-01
Budget End
2010-11-30
Support Year
5
Fiscal Year
2009
Total Cost
$348,689
Indirect Cost
Name
Rockefeller University
Department
Genetics
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Pomeranz, Lisa E; Ekstrand, Mats I; Latcha, Kaamashri N et al. (2017) Gene Expression Profiling with Cre-Conditional Pseudorabies Virus Reveals a Subset of Midbrain Neurons That Participate in Reward Circuitry. J Neurosci 37:4128-4144
Ekstrand, Mats I; Nectow, Alexander R; Knight, Zachary A et al. (2014) Molecular profiling of neurons based on connectivity. Cell 157:1230-42
PĂ©rez, Cristian A; Stanley, Sarah A; Wysocki, Robert W et al. (2011) Molecular annotation of integrative feeding neural circuits. Cell Metab 13:222-32
Stanley, Sarah; Pinto, Shirly; Segal, Jeremy et al. (2010) Identification of neuronal subpopulations that project from hypothalamus to both liver and adipose tissue polysynaptically. Proc Natl Acad Sci U S A 107:7024-9
Ekstrand, Mats I; Enquist, L W; Pomeranz, Lisa E (2008) The alpha-herpesviruses: molecular pathfinders in nervous system circuits. Trends Mol Med 14:134-40