RESEARCH ARTICLE
Differential Effects of C1qa Ablation on Glaucomatous Damage in Two Sexes in DBA/ 2NNia Mice Ruma Kumari1,3☯*, Konstantin Astafurov1,3☯, Alina Genis2,3, John Danias1,2,3* 1 Department of Cell Biology, State University of New York (SUNY) Downstate Medical Center, Brooklyn, New York, United States of America, 2 Department of Ophthalmology, State University of New York (SUNY) Downstate Medical Center, Brooklyn, New York, United States of America, 3 State University of New York (SUNY) Eye Institute, Brooklyn, New York, United States of America ☯ These authors contributed equally to this work. *
[email protected] (RK);
[email protected] (JD)
Abstract OPEN ACCESS
Purpose
Citation: Kumari R, Astafurov K, Genis A, Danias J (2015) Differential Effects of C1qa Ablation on Glaucomatous Damage in Two Sexes in DBA/2NNia Mice. PLoS ONE 10(11): e0142199. doi:10.1371/ journal.pone.0142199
To determine the sex and age-related effects of C1qa ablation on retinal ganglion cell (RGC) and optic nerve (ON) axonal loss in a mouse model of glaucomatous neurodegeneration.
Editor: Glyn Chidlow, Hanson Institute, AUSTRALIA
Methods
Received: April 9, 2015
Congenic C1qa mice were generated in the DBA/2NNia background. Female and male knockout (-/-), heterozygous (+/-), and wild type (+/+) mice were aged up to 14 months and IOPs were recorded in a subset of animals. Retinas of mice from all three groups at 5–6, 9–10 and 11–13 months of age were flat-mounted after retrograde labeling with Fluorogold. Imaged retinas were scored (RGC score) semi-quantitatively on a 10 point scale by two independent observers. A subset of retinas and optic nerves were also used for measurement of total number of RGCs. Semi-thin sections of ON were imaged and graded (ON score) for the amount of axonal damage semi-quantitatively, by two masked observers. Analysis of covariance (ANCOVA) was used for statistical comparisons. Microglial cells in flat-mounted retinas of 5–6 month old C1qa -/- and C1qa +/+ mice were used for assessment of microglial activation utilizing morphological criteria.
Accepted: October 18, 2015 Published: November 6, 2015 Copyright: © 2015 Kumari et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper. Funding: This work received financial support from the following sources: grant # 5R01EY015224 from National Eye Institute to JD; Unrestricted challenge grant to the Dept. of Ophthalmology of SUNY Downstate from the Research to Prevent Blindness. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist.
Results Female C1qa -/- mice had significantly higher IOP (p8), moderate (RGC scores >4 and 8) or severe (RGC scores 4) damage was also not significantly different between the three genotypes at 5–6 months of age (p>0.28, Fisher exact test, Fig 4A). At 9–10 months of age RGC scores of C1qa -/- and C1qa +/- animals were significantly higher compared to those of C1qa +/+ animals (p0.7 for male and female mice respectively, ANCOVA, Fig 5A and 5B). In 9–10 month old male mice, mean ON scores were significantly lower in C1qa -/and C1qa +/- animals compared to C1qa +/+ animals (p
