Abstract
Purpose Despite popularity of optical coherence tomography (OCT) in glaucoma studies, it’s unclear how well OCT-derived metrics compare to traditional measures of retinal ganglion cell (RGC) abundance. Here, Diversity Outbred (J:DO) mice are used to directly compare ganglion cell complex (GCC)-thickness measured by OCT to metrics of retinal anatomy measured ex vivo with retinal wholemounts or optic nerve cross sections.
Methods J:DO mice (n = 48) underwent OCT and fundoscopic exams, with GCC-thickness measured using automated segmentation. Following euthanasia, RGC axons were quantified from para- phenylenediamine-stained optic nerve cross sections and RGC somas from BRN3A-immunolabeled retinal wholemounts with total cellularity assessed by TO-PRO or hematoxylin nuclear staining.
Results J:DO tissues lacked overt disease. GCC-thickness (62.4 ± 3.7 µm), RGC abundance (3,097 ± 515 BRN3A+ nuclei/mm2; 45,533 ± 9,077 axons), and total inner retinal cell abundance (6,952 ± 810 nuclei/mm2) varied broadly. GCC-thickness correlated significantly to RGC somal density (r = 0.46) and axon number (r = 0.49), whereas total inner retinal cellularity did not. Retinal area (20.3 ± 2.4 mm2) and optic nerve (0.09 ± 0.02 mm2) cross-sectional area varied widely. Sex did not significantly influence any of these metrics. In bilateral comparisons, GCC-thickness (r = 0.89), inner retinal cellularity (r = 0.47), and RGC axon abundance (r = 0.72) all correlated significantly.
Conclusions Amongst outbred mice with widely variable phenotypes, OCT-derived measurements of GCC thickness correlate significantly to RGC abundance and axon number. The extensive phenotypic variability exhibited by J:DO mice make them a powerful resource for studies of retinal anatomy using quantitative genetics.
Competing Interest Statement
Disclosure: A. Hedberg-Buenz, none; C.J. van der Heide, none; K.J. Meyer, none; D. Soukup, none; W. Deng, none; K. Lee, none; H. Mercer, none; M. Kettelson, Digital Diagnostics, Inc.; D. Pellack, none; K. Wang, none; M.K. Garvin, is the co-inventor on a US patent related to the approach used to segment retinal layers; she has personally waived all financial rights to said patent, but the University still has rights; M.D. Abramoff, is the inventor on patents and patent applications of artificial intelligence and machine learning algorithms for diagnosis and treatment and is a Founder, CEO, employee of, and investor in Digital Diagnostics Inc, Coralville, Iowa, USA.; and M.G. Anderson, none.