Multispectral labeling technique to map many neighboring axonal projections in the same tissue.
We describe a method to map the location of axonal arbors of many individual neurons simultaneously via the spectral properties of retrogradely transported dye-labeled vesicles. We inject overlapping regions of an axon target area with three or more different colored retrograde tracers. On the basis of the combinations and intensities of the colors in the individual vesicles transported to neuronal somata, we calculate the projection sites of each neuron's axon. This neuronal positioning system (NPS) enables mapping of many axons in a simple automated way. In our experiments, NPS combined with spectral (Brainbow) labeling of the input to autonomic ganglion cells showed that the locations of ganglion cell projections to a mouse salivary gland related to the identities of their preganglionic axonal innervation. NPS could also delineate projections of many axons simultaneously in the mouse central nervous system.
1] Center for Brain Science, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA. [3] Department of Medical Neurobiology, Institute for Medical Research Israel Canada Faculty of Medicine, The Hebrew University, Jerusalem, Israel. [4] The Edmond and Lily Safra Center for Brain Sciences, The Hebrew University, Jerusalem, Israel.
Shlomo
S
Tsuriel
1] Department of Medical Neurobiology, Institute for Medical Research Israel Canada Faculty of Medicine, The Hebrew University, Jerusalem, Israel. [2] The Edmond and Lily Safra Center for Brain Sciences, The Hebrew University, Jerusalem, Israel.
Sagi
S
Gudes
1] Center for Brain Science, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
Ryan W
RW
Draft
1] Department of Medical Neurobiology, Institute for Medical Research Israel Canada Faculty of Medicine, The Hebrew University, Jerusalem, Israel. [2] The Edmond and Lily Safra Center for Brain Sciences, The Hebrew University, Jerusalem, Israel.
Alexander M
AM
Binshtok
1] Center for Brain Science, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
Jeff W
JW
Lichtman
D000818:Animals; D001369:Axons; D001931:Brain Mapping / Q000379:methods; D002540:Cerebral Cortex / Q000166:cytology*; D003196:Computer Graphics; D005726:Ganglia, Parasympathetic / Q000166:cytology*; D007091:Image Processing, Computer-Assisted; D051379:Mice; D009434:Neural Pathways / Q000502:physiology; D009474:Neurons / Q000166:cytology*; D013194:Staining and Labeling / Q000379:methods*; D013788:Thalamus / Q000166:cytology*