For inactivation

of AlstR-expressing neurons, the peptide ligand AL (Ser-Arg-Pro-Tyr-Ser-Phe-Gly-Leu-NH2) was applied by perfusion. Organotypic brain slice cultures were used for testing SADΔG-GFP-rtTA. After biolistic transfection of both pCMMP-TVA800 and pTetO-CMVmin-Histone2B-mCherry-F2A-B19G, slices were infected with EnvA-SADΔG-GFP-rtTA and maintained in the selleck products absence or presence of dox (1.0 μg/ml). For testing SADΔG-GFP-ERT2CreERT2, HEK293t cells were transfected with the Cre-dependent plasmid pCALNL-DsRed, infected with the rabies virus, and maintained in the absence or presence of 4-HOT (1.0 μM). For testing SADΔG-FLPo-DsRedX, HeLa cells stably expressing a frt-STOP-frt nuclear-localized LacZ cassette were infected with the virus and then processed R428 in vitro for X-gal staining. Full methods are available in Supplemental Information. We thank I.R. Wickersham and J. Choi for helpful discussions; K.D. Roby, M. De La Parra, and K. von Bochmann for technical assistance; members of the Callaway laboratory for stimulating discussions; K.K. Conzelmann for the

BSR T7/5 cell line; O. Britz and M. Goulding for the HeLa cells expressing frt-STOP-frt-nLacZ; I.M. Verma for HIV lentivirus packaging plasmids; X. Wu for the pNLST7; R. Tsien for the mCherry plasmid; K. Deisseroth for the ChR2-mCherry plasmid; L.L. Looger for the GCaMP3 plasmid; and C.L. Cepko for the pCAG-ERT2CreERT2 and pCALNL-DsRed. F.O. is thankful to N. Osakada for constant encouragement and support. We are grateful for support from the National Institutes of Health (MH063912, NS069464, and EY010742: E.M.C.), the Kavli Institute for Brain and Mind at University of California San Diego (E.M.C.), the Japan Society for the Promotion of Science (F.O.), the Kanae Foundation for the Promotion of Medical Science (F.O.), the Uehara Memorial Foundation (F.O.), and the Naito Foundation (F.O.). “
“Functional neural circuits consist of precise connectivity between specific sets of neurons. The assembly of such circuitry often requires that axons bypass numerous targets before

selectively terminating in just one or a few specific targets. Over the last century, much progress has been made in understanding however how axons undergo directed growth and pathfinding and how they form topographic maps (Sperry, 1963, Dickson, 2002 and Feldheim and O’Leary, 2010). How mammalian axons identify which targets to innervate, however, remains poorly understood. The axonal connections formed by the eyes with the brain are an attractive model for exploring mechanisms of axon-target recognition in the mammalian CNS. Retinal ganglion cells (RGCs) are the output neurons of the eye and they are divided into ∼20 different types. Each RGC type encodes a different quality of the visual scene, such as brightness, direction of motion or edges (Masland, 2001 and Berson, 2008), and sends that information to a limited number of retinorecipient targets that in turn regulate specific aspects of perception and behavior.