Kenneth M P Yee1, Edward P Feener2, Michele Madigan3, Nicholas J Jackson4, Ben-Bo Gao2, Fred N Ross-Cisneros5, Jan Provis6, Lloyd Paul Aiello7, Alfredo A Sadun8, J Sebag1. 1. VMR Institute for Vitreous Macula Retina, Huntington Beach, California, United States 2Doheny Eye Institute, Los Angeles, California, United States. 2. Beetham Eye Institute, Joslin Diabetes Center, Boston, Massachusetts, United States. 3. School of Optometry and Vision Science, University of New South Wales, Sydney, Australia 5Save Sight Institute, University of Sydney, Sydney, Australia. 4. Department of Medicine Statistics Core, David Geffen School of Medicine at UCLA, Los Angeles, California, United States. 5. Doheny Eye Institute, Los Angeles, California, United States. 6. John Curtin School of Medical Research, Canberra, Australia 8Australian National University, Canberra, Australia. 7. Beetham Eye Institute, Joslin Diabetes Center, Boston, Massachusetts, United States 9Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States. 8. Doheny Eye Institute, Los Angeles, California, United States 10Department of Ophthalmology, David Geffen School of Medicine at UCLA, Los Angeles, California, United States.
Abstract
PURPOSE: The proteomic profile of vitreous from second-trimester human embryos and young adults was characterized using mass spectrometry and analyzed for changes in protein levels that may relate to structural changes occurring during this time. This vitreous proteome was compared to previous reports to confirm proteins already identified and reveal novel ones. METHODS: Vitreous from 17 human embryos aged 14 to 20 weeks gestation (WG) and from a 12-, a 14-, a 15-, and a 28-year-old was individually analyzed using tandem mass spectrometry-based proteomics. Peptide spectral count associations with embryonic age were assessed using a general linear model of fold changes and Spearman's rank correlation. Differences between embryonic and young adult vitreous proteomes were also compared. Immunohistochemistry was used to evaluate three proteins in five additional fetal (10-18 WG) human eyes. RESULTS: There were 1217 proteins identified in fetal and young adult human vitreous, 206 after quantile normalization and variance filtering. In embryos, the peptide counts of 37 proteins changed significantly from 14 to 20 WG: 75.7% increased, 24.3% decreased. Immunohistochemistry confirmed the absence of clusterin and cadherin in 10 and 14 WG eyes and their presence at 18 WG. Comparing embryonic to young adult vitreous, 47 proteins were significantly higher or lower. A total of 768 proteins not previously identified in the literature are presented. CONCLUSIONS: Proteins previously unreported in the human vitreous were identified. The human vitreous proteome undergoes significant changes during embryogenesis and young adulthood. A number of protein levels change considerably during the second trimester, with the majority decreasing.
PURPOSE: The proteomic profile of vitreous from second-trimester human embryos and young adults was characterized using mass spectrometry and analyzed for changes in protein levels that may relate to structural changes occurring during this time. This vitreous proteome was compared to previous reports to confirm proteins already identified and reveal novel ones. METHODS: Vitreous from 17 human embryos aged 14 to 20 weeks gestation (WG) and from a 12-, a 14-, a 15-, and a 28-year-old was individually analyzed using tandem mass spectrometry-based proteomics. Peptide spectral count associations with embryonic age were assessed using a general linear model of fold changes and Spearman's rank correlation. Differences between embryonic and young adult vitreous proteomes were also compared. Immunohistochemistry was used to evaluate three proteins in five additional fetal (10-18 WG) human eyes. RESULTS: There were 1217 proteins identified in fetal and young adult human vitreous, 206 after quantile normalization and variance filtering. In embryos, the peptide counts of 37 proteins changed significantly from 14 to 20 WG: 75.7% increased, 24.3% decreased. Immunohistochemistry confirmed the absence of clusterin and cadherin in 10 and 14 WG eyes and their presence at 18 WG. Comparing embryonic to young adult vitreous, 47 proteins were significantly higher or lower. A total of 768 proteins not previously identified in the literature are presented. CONCLUSIONS: Proteins previously unreported in the human vitreous were identified. The human vitreous proteome undergoes significant changes during embryogenesis and young adulthood. A number of protein levels change considerably during the second trimester, with the majority decreasing.
Authors: Susan M Keezer; Susan E Ivie; Henry C Krutzsch; Anita Tandle; Steven K Libutti; David D Roberts Journal: Cancer Res Date: 2003-10-01 Impact factor: 12.701