Postsynaptic cell type and synaptic distance do not determine efficiency of monosynaptic rabies virus spread measured at synaptic resolution
- Systems Neurobiology Laboratories, The Salk Institute for Biological Studies, United States
- Neuroscience Graduate Program, University of California, San Diego, United States
- Medical Scientist Training Program, University of California, San Diego, United States
- Allen Institute for Brain Science, United States
Figures
Figure 1
RVdG-PSD95GFP-SynPhRFP allows simultaneous fluorescent labeling of pre- and postsynaptic densities.
(A) Schematic of RVdG-PSD95GFP-SynPhRFP viral construct design. Two transgenes were inserted into the G locus of the rabies genome. One encodes a presynaptically targeted fluorescent fusion protein, …
-
Figure 1—source data 1
Plasmid sequence for RVdG-PSD95GFP-SynPhRFP construct.
Plasmid sequence for a new genetically modified rabies virus that labels presynaptic terminals with synaptophysin-RFP (SynPhRFP) and excitatory postsynaptic densities with postsynaptic density-95-GFP (PSD95GFP).
- https://cdn.elifesciences.org/articles/89297/elife-89297-fig1-data1-v1.zip
Figure 2
Experimental design for control experiments omitting oG.
(A) Schematic illustration of experimental design and timeline for control experiments without glycoprotein. Nr5a1Cre mice were injected in V1 with a mixture of AAV-CIAO-TVA and AAV-FLEX-smFP-myc. …
Figure 3
Efficiency of transsynaptic spread from excitatory L4 Nr5a1+ starter cells to excitatory inputs.
(A) Schematic illustration of experimental design and timeline for monosynaptic rabies tracing. Nr5a1Cre mice were injected in V1 with a mixture of AAV-CIAO-TVA, AAV-FLEX-H2BBFP-oG, and …
Figure 4
Efficiency of transsynaptic spread from inhibitory starter cells to excitatory inputs.
(A) Representative example images of V1 injection site for SstCre mouse line (top) and VipCre mouse line (bottom), obtained using widefield fluorescence microscopy at 10×. Scale bar = 200 µm. (B–E) …
Figure 5
Quantification from electron micrographic reconstructions of: (A) shared inputs between layer 4 excitatory neurons and (B) the number of synapses per excitatory neuron to excitatory neuron connection.
(A) The percentage of inputs to a layer 4 excitatory neuron that provide common input to another layer 4 excitatory neuron (percentage of neurons connected) at increasing distances between layer 4 …
Figure 6
Relationships between synaptic fraction (SF), input fraction (IF), and unitary synaptic efficiency (U) depend on synapses per input neuron (M) and proportions of neurons with differences in M.
(A and C) SF against the ratio (R) of neurons with M values of 1 versus either 3 (A) or 2 (C) for different values of U. (B and D) Ratio of IF versus SF against the R of neurons with M values of 1 …
Author response image 1
Additional files
-
MDAR checklist
- https://cdn.elifesciences.org/articles/89297/elife-89297-mdarchecklist1-v1.pdf
-
Source data 1
Raw data of pre-and post-synaptic densities for Figures 2—4.
Raw data includes counts for Nr5a1, Som, and vasoactive intestinal peptide (Vip) starter cells and control experiments omitting oG.
- https://cdn.elifesciences.org/articles/89297/elife-89297-data1-v1.xlsx
