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      Changes of osteopontin in the aqueous humor of the DBA2/J glaucoma model correlated with optic nerve and RGC degenerations.

      Investigative ophthalmology & visual science

      metabolism, pathology, Aqueous Humor, Blotting, Western, Cell Survival, drug effects, Cytokines, Animals, Disease Models, Animal, Enzyme-Linked Immunosorbent Assay, Glaucoma, In Situ Hybridization, Intraocular Pressure, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Nerve Degeneration, drug therapy, Optic Nerve Diseases, Organ Culture Techniques, Osteopontin, genetics, pharmacology, RNA, Messenger, Retinal Ganglion Cells, Aging, physiology

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          To identify age-dependent regulated aqueous humor (AH) factors in DBA2/J (D2J) mice and to correlate them with optic nerve degeneration and intraocular pressure (IOP) by population and individual analysis. AH samples of D2J mice aged 2 (n = 3), 7 (n = 5), and 10 months (n = 14) were analyzed by mouse cytokine antibody array. Ten-month samples were classified into eyes with (D2J+) or without (D2J-) optic neuropathy. Ten-month-old C57/Bl6 (B6; n = 13) and DBA2/Rj (D2Rj; n = 15) mice served as controls. IOP was recorded from 2 to 10 months. Individual AH osteopontin (OPN) was determined in 31 D2J eyes (10 months) and was correlated with optic neuropathy and IOP. OPN mRNA was detected by in situ hybridization. OPN blood plasma content of D2J and B6 was monitored from 8 to 10 months. Effect of OPN on cell survival in the ganglion cell layer (GCL) or metabolism was tested in ex vivo-cultured D2Rj eyes and murine neuronal precursors. In array analysis, OPN was detected in 10-month-old D2J mice only. They significantly differed between D2J- and D2J+ (P = 0.006). By Western blot analysis, a sevenfold OPN increase in D2J+ was determined compared with B6. Individual analysis confirmed the positive correlation of OPN with optic neuropathy. IOP was not correlated with OPN. OPN blood plasma contents steadily increased with age in D2J. OPN(+) cells were detected within the ciliary body of D2J, and OPN(+) RGCs were ≈30% reduced. OPN treatment inhibited cell degeneration within the GCL in ex vivo-cultured D2Rj eyes and increased the metabolic activity of neuronal precursor cells. OPN is an age-dependent increased AH factor associated with degeneration of the optic nerve in D2J mice. By modulating the metabolism of neuronal cells, deregulated levels of OPN could be involved in degenerative processes affecting RGCs or optic nerve axons in the D2J model.

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