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Ayman Aljohani, Yenifer S Guerra, Sanjoy K Bhattacharya; Differential Comparison of Trabecular Meshwork Ceramides from Control and Glaucomatous Donors. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5705.
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To compare the ceramides of the Trabecular Meshwork (TM) from control and primary open angle glaucoma (POAG) donors by mass spectrometric shotgun lipidomics.
The human TM samples were collected adhering to tenets of declaration of Helsinki under IRB approved protocols from control and POAG donors of diverse race and both genders with mean age of 55.9 ±14.2 years (n=20 each). TM Ceramides were extracted using the Bligh and Dyer method, re-suspended in Isoproyl, Acetonitrile solution and analyzed on a TSQ Quantum Access Max instrument in negative mode. Neutral loss scan (NLS) for ceramides (Cer; product m/z of 256.2) were performed using direct infusion. Ratiometric quantification was performed for m/z range of 200-1000. The ceramide species of control and glaucoma patients were identified using MZmine 2.9 version and database from lipidmaps.org. The profiles were compared using in-house written softwares. Further confirmatory analyses were performed on a high resolution Q-exactive mass spectrometer.
Overall total amount of ceramides in POAG normalized to protein amounts were always less than that in controls. No ceramide species was found unique between control and POAG, however 62 ceramides were common between these two groups. The ceramides: Cer (d18:0/14:0), Cer(d18:0/16:0) and Cer(d18:2/16:0) undergoes a significant decrease in POAG compared to controls, while about two species [Cer(d18:0/13:0); Cer(d18:0/24:1(15Z))] show a marginal increase in POAG.
Lower amounts of ceramides were found in TM of POAG compared to controls. This was confirmed by high resolution mass spectrometry. This is likely to contribute to reduced membrane fluidity and likely result in stiffness to TM cells in glaucoma.
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