Survival: RGC survival was evaluated at ten weeks soon after the induction of elevated IOP. There was a considerable decrease in the RGC quantity with age in the manage fellow eyes: It dropped from 1049?6 RGC/mm two at three months to 955?7.6 at six months and 725?two RGC/mm two at 18 months (n=4? for each age group,Molecular Vision 2013; 19:2011-2022 molvis.org/molvis/v19/2011?2013 Molecular Visionp=0.002, analysis of variance [ANOVA], Figure 2A). Furthermore, elevated IOP induced a significant loss of RGCs in every age group: The number decreased from 669?23 RGC/mm 2 at three months to 486?14 RGC/mm2 at six months and 189?six.5 RGC/mm 2 at 18 months (n=4?, p=0.048, ANOVA; Figure 2A). Hence, there was greater glaucomatous RGC loss with age beginning with a 35.eight ?11.5 loss at 3 months of age to a 39.four ?11.7 loss at six months and progressing to a 74 ?six loss at 18 months (n=4?, p=0.055, ANOVA, Figure 2B). This age-related progression in RGC loss occurred under similar IOP levels. Quantitative polymerase chain reaction array for apoptosis in aged glaucomatous eyes: PCR array results revealed potential gene expression changes which can shed light around the causes for the enhanced susceptibility of aged RGCs to injury. Genes that had been up- or downregulated with at least a twofold adjust are presented in bold in Table 2. Twenty genes were upregulated in the 3-month-old rats, 8 genes within the 13 month olds, and 12 in the 18 month olds. Downregulation was observed in 16 genes within the 3 month olds, 29 genes in the 13 month olds, and 4 genes within the 18 month olds. The upregulated genes in the 3-month-old group included the Bcl-2 p38 MAPK Inhibitor Synonyms family members (Bcl2, Bcl2l1), NLR loved ones apoptosis inhibitory protein 2 (Naip2), caspase family (four, six, and 7), Fas PLK1 Inhibitor Purity & Documentation apoptotic inhibitory molecule (Faim), the tumor necrosis factor (TNF) loved ones (Tnfrsf1a, Tnfrsf1b, and Traf4), and Tp53bp2. The downregulated genes have been members of the caspase household (8, 14, and Casp8ap2), TNF household (Tnf, Tnfrsf10b, Tnfrsf11b), Tp63, and Tp73. The upregulated genes inside the 13-month-old group had been proapoptotic genes that included TNF family members (Tnf, Tnfrsf11b, Tnfsf10, and Fas) and caspase family members (4 and 12; Table 2). The downregulated genes have been members from the Bcl-2 loved ones, various caspase members of the family (1, 14, 7, and eight), and tumor protein p53 (p53) family members (Table 2). The upregulated genes inside the 18-month-old group also incorporated TNF members of the family (Tnf, Tnfrsf1a), several caspase members of the family (1 and four) and bcl-2. Amongst the downregulated genes were DNA fragmentation element, beta subunit (DffB), and p53. Validation of reverse transcription polymerase chain reaction: The expressions of chosen proapoptotic and prosurvival genes have been determined working with RT CR to validate the PCR array outcomes (Figure 3). Essentially the most crucial (and unexpected) obtaining was the difference between young and old rats in expression levels from the two significant prosurvival genes, IAP and XIAP. IAP-1 mRNA levels increased by 111.7?.5 within the 3-month glaucomatous eyes in comparison to the fellow manage eyes (n=5, p=0.0002), however it decreased by 31.0?.9 inside the 15-month-old rats (n=6, p=0.002; Figure 3A). AnotherIAP family member, the prosurvival XIAP gene, enhanced by 53.0?eight.2 within the 3-month-old glaucomatous eyes (n=6, p=0.04), but decreased drastically (by 41.6?.2 ) within the 15-month-old eyes (n=7, p=0.04; Figure 3B). There have been no modifications in P53 mRNA levels within the 3-month-old glaucomatous rats; nevertheless, there was a trend toward decline in the 15- m.