Analysis of Retinal Vascular Plexuses and Interplexus Connections
The retina is a highly organized neural tissue consisting of three neural layers and two synaptic layers. Blood vessels that nourish the mouse and human neural retina mirror this organization consisting of three plexus layers, or plexuses, that run parallel within the retina, connected by interplexus vessels to create a closed vascular network. Here, we describe a methodology to describe this organization that can be used to interrogate factors mediating retinal vessel patterning including: coverage of the vascular plexuses, branching and orientation of the interplexus connections, and digital reconstruction of the retinal vasculature to measure vessel length and density. The methodology focuses on the mouse retina, but can easily be adapted to study retinal vessels of other species.
Key wordsDevelopment Protocol Assay Retinopathy Artery Vein Capillary Angiogenesis VEGF Macular degeneration Vasculopathy
- 3.Simmons AB, Merrill MM, Reed JC, Deans MR, Edwards MM, Fuerst PG (2016) Defective angiogenesis and intraretinal bleeding in mouse models with disrupted inner retinal lamination. Invest Ophthalmol Vis Sci 57(4):1563–1577. https://doi.org/10.1167/iovs.15-18395 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Usui Y, Westenskow PD, Kurihara T, Aguilar E, Sakimoto S, Paris LP, Wittgrove C, Feitelberg D, Friedlander MS, Moreno SK, Dorrell MI, Friedlander M (2015) Neurovascular crosstalk between interneurons and capillaries is required for vision. J Clin Invest 125(6):2335–2346. https://doi.org/10.1172/JCI80297 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Shen W, Fruttiger M, Zhu L, Chung SH, Barnett NL, Kirk JK, Lee S, Coorey NJ, Killingsworth M, Sherman LS, Gillies MC (2012) Conditional Mullercell ablation causes independent neuronal and vascular pathologies in a novel transgenic model. J Neurosci 32(45):15715–15727. https://doi.org/10.1523/JNEUROSCI.2841-12.2012 CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Scott A, Powner MB, Gandhi P, Clarkin C, Gutmann DH, Johnson RS, Ferrara N, Fruttiger M (2010) Astrocyte-derived vascular endothelial growth factor stabilizes vessels in the developing retinal vasculature. PLoS One 5(7):e11863. https://doi.org/10.1371/journal.pone.0011863 CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Linkert M, Rueden CT, Allan C, Burel JM, Moore W, Patterson A, Loranger B, Moore J, Neves C, Macdonald D, Tarkowska A, Sticco C, Hill E, Rossner M, Eliceiri KW, Swedlow JR (2010) Metadata matters: access to image data in the real world. J Cell Biol 189(5):777–782. https://doi.org/10.1083/jcb.201004104 CrossRefPubMedPubMedCentralGoogle Scholar