Fig 1.
Labeling excitatory synaptic input on retinal ganglion cells.
The synaptic sites were labeled with PSD-95-GFP that was delivered into retinal ganglion cells by in-vivo transfection with AAV2 (green). The dendrites were counterstained with an antibody against neurofilament (SMI32, magenta).
Fig 2.
Labeling synaptic input on individual retinal ganglion cells in non-glaucomatous retinas.
A, Excitatory synaptic inputs were labeled by AAV2-mediated transfection of ganglion cells with PSD-95-GFP, leading to a punctuate labeling. B, To assign synaptic puncta to individual ganglion cells, the cytoskeleton of large ganglion cells was labeled with an antibody against neurofilament. C, Overlay of A and B. D, Digital representation of the dendritic arbor with assigned synaptic input sites.
Fig 3.
Retina whole-mounts stained for the axonal marker SMI32 (neurofilament).
A, D2.Gpnmb+ mice with no axon loss. B, retina from a 11 month old DBA/2J mouse with severe loss of ganglion cells and axons. C. Control retina from a C57bl/6 mouse. D. Retina from a C57bl/6 mouse 7 days after optic nerve crush.
Fig 4.
Labeling of synaptic inputs to ganglion cells in glaucomatous DBA/2J retinas.
A, Digital representation of a non-glaucomatous (D2.Gpnmb+) retina. B, Higher magnification of the boxed area in panel A, showing the density of excitatory synaptic inputs to a higher-order dendrite. C & E, Two examples of ganglion cells from glaucomatous DBA/2J mice. The dendritic remodeling is obvious. D & F, Higher magnification views of the boxed areas in panels C and E, respectively. Overall synaptic density is lower than in the non-glaucomatous ganglion cells.
Table 1.
Mean Overall Linear Density and Complexity Values across Experimental Conditions.
Mean values for overall linear density and complexity are shown with associated standard errors of the mean (n = 10 cells for each condition). Comparisons between experimental conditions and corresponding controls are indicated by significance values generated via two-tailed T-tests.
Fig 5.
Correlation of synaptic density with complexity of the dendritic arbor in glaucomatous DBA/2J and control D2.Gpnmb+ retinas.
A, Scatterplot of overall linear density of PSD-95 puncta reveals a weak correlation with the complexity of the dendritic arbor as measured by the number of non-terminal nodes. B, the correlation was repeated for the densest branch.
Fig 6.
Labeling of synaptic inputs to ganglion cells after optic nerve crush.
A, Normal cell from the contralateral (uncrushed) eye. B, Higher magnification of the boxed area in panel A. C & E, Two examples of ganglion cells one week after optic nerve crush. Note the decreased complexity of the dendritic arbors. D & F, Higher magnification of the boxed areas in panels C and E.
Fig 7.
Correlation of synaptic density with complexity of the dendritic arbor in control C57bl/6 mice and in C57bl/6 mice after optic nerve crush.
A, Scatterplot of synaptic density with complexity of the dendritic arbor in control retinas, and retinas after optic nerve crush. B, correlation of the synaptic density with complexity of the dendritic arbor for the densest branch.