a dual infection pseudorabies virus conditional reporter approach to identify projections to collateralized neurons in complex neural circuits假狂犬病病毒双重感染条件记者方法识别预测抵押神经元在复杂的神经回路.pdfVIP

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a dual infection pseudorabies virus conditional reporter approach to identify projections to collateralized neurons in complex neural circuits假狂犬病病毒双重感染条件记者方法识别预测抵押神经元在复杂的神经回路.pdf

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a dual infection pseudorabies virus conditional reporter approach to identify projections to collateralized neurons in complex neural circuits假狂犬病病毒双重感染条件记者方法识别预测抵押神经元在复杂的神经回路

A Dual Infection Pseudorabies Virus Conditional Reporter Approach to Identify Projections to Collateralized Neurons in Complex Neural Circuits 1 . 2. 2 1 1 2 J. Patrick Card * , Oren Kobiler , Ethan B. Ludmir , Vedant Desai , Alan F. Sved , Lynn W. Enquist 1 Department of Neuroscience, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America, 2 Department of Molecular Biology and the Princeton Neuroscience Institute, Princeton University, Princeton, New Jersey, United States of America Abstract Replication and transneuronal transport of pseudorabies virus (PRV) are widely used to define the organization of neural circuits in rodent brain. Here we report a dual infection approach that highlights connections to neurons that collateralize within complex networks. The method combines Cre recombinase (Cre) expression from a PRV recombinant (PRV-267) and Cre-dependent reporter gene expression from a second infecting strain of PRV (PRV-263). PRV-267 expresses both Cre and a monomeric red fluorescent protein (mRFP) fused to viral capsid protein VP26 (VP26-mRFP) that accumulates in infected cell nuclei. PRV-263 carries a Brainbow cassette and expresses a red (dTomato) reporter that fills the cytoplasm. However, in the presence of Cre, the dTomato gene is recombined from the cassette, eliminating expression of the red reporter and liberating expression of either yellow (EYFP) or cyan (mCerulean) cytoplasmic reporters. We conducted proof-of-principle experiments using a well-characterized model in which separate injection of recombinant viruses into the left and right kidneys produces infection of neurons in the renal preautonomic network. Neurons dedicated to one kidney expresse