The effects of age on mitochondria, axonal transport, and axonal degeneration after chronic IOP elevation using a murine ocular explant model

• Published in: Experimental eye research 2018-07, Vol.172, p.78-85
• Main Authors: Kimball, Elizabeth C., Jefferys, Joan L., Pease, Mary E., Oglesby, Ericka N., Nguyen, Cathy, Schaub, Julie, Pitha, Ian, Quigley, Harry A.
• Format: Article
• Language: English
• Subjects: Age Factors; Animals; Axonal Transport - physiology; Axons; Axons - pathology; Bacterial Proteins - metabolism; Chronic Disease; Disease Models, Animal; Explant; Glaucoma; Green Fluorescent Proteins - metabolism; Intraocular Pressure - physiology; Luminescent Proteins - metabolism; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Confocal; Mitochondria; Mitochondria - pathology; Mouse; Ocular Hypertension - pathology; Optic Disk - pathology; Retinal ganglion cell; Retinal Ganglion Cells - metabolism; Retinal Ganglion Cells - pathology; Sclera; Tonometry, Ocular; Transport block
• ISSN: 0014-4835; 1096-0007
• DOI: 10.1016/j.exer.2018.04.001

The purpose of this study was to compare younger and older mice after chronic intraocular pressure (IOP) elevation lasting up to 4 days with respect to mitochondrial density, structure, and movement, as well as axonal integrity, in an ex vivo explant model. We studied 2 transgenic mouse strains, both on a C57BL/6J background, one expressing yellow fluorescent protein (YFP) in selected axons and one expressing cyan fluorescent protein (CFP) in all mitochondria. Mice of 4 months or 14 months of age were exposed to chronic IOP by anterior chamber microbead injection for 14 h, 1, 3, or 4 days. The optic nerve head of globe--optic nerve explants were examined by laser scanning microscopy. Mitochondrial density, structure, and movement were quantified in the CFP explants, and axonal integrity was quantified in YFP explants. In control mice, there was a trend towards decreased mitochondrial density (# per mm2) with age when comparing younger to older, control mice, but this was not significant (1947 ± 653 vs 1412 ± 356; p = 0.19). Mitochondrial density decreased after IOP elevation, significantly, by 31%, in younger mice (p = 0.04) but trending towards a decrease, by 22%, in older mice (p = 0.82) compared to age matched controls. Mitochondrial mean size was not altered after chronic IOP elevation for 14 h or more (p ≥ 0.16). When assessing mitochondrial movement, in younger mice, 5% were mobile at any given time; 4% in the anterograde direction and 1% retrograde. In younger untreated tissue, only 75% of explants had moving mitochondria (mean = 15.8 moving/explant), while after glaucoma induction only 24% of explants had moving mitochondria (mean = 4.2 moving/explant; difference from control, p = 0.03). The distance mitochondria traveled in younger mice was unchanged after glaucoma exposure, but in older glaucoma explants the distance traveled was less than half of older controls (p