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Differential Expression of Vitreous Proteins in Young and Mature New Zealand White Rabbits.

Liu Y, Bouhenni RA, Dufresne CP, Semba RD, Edward DP - PLoS ONE (2016)

Bottom Line: Our results showed that 231, 375, 273 and 353 proteins were identified in the YA, YP, MA and MP respectively.Fifteen proteins were significantly differentially expressed between YA and YP, and 11 between MA and MP.Western blotting confirmed the differential expression of carbonic anhydrase III, alpha crystallin B and beta crystallin B2.

View Article: PubMed Central - PubMed

Affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, United States of America.

ABSTRACT
Different anatomical regions have been defined in the vitreous humor including central vitreous, basal vitreous, vitreous cortex, vitreoretinal interface and zonule. In this study we sought to characterize changes in the proteome of vitreous humor (VH) related to compartments or age in New Zealand white rabbits (NZW). Vitreous humor was cryo-collected from young and mature New Zealand white rabbit eyes, and dissected into anterior and posterior compartments. All samples were divided into 4 groups: Young Anterior (YA), Young Posterior (YP), Mature Anterior (MA) and Mature Posterior (MP) vitreous. Tryptic digests of total proteins were analyzed by liquid chromatography followed by tandem mass spectrometry. Spectral count was used to determine the relative protein abundances and identify proteins with statistical differences between compartment and age groups. Western blotting was performed to validate some of the differentially expressed proteins. Our results showed that 231, 375, 273 and 353 proteins were identified in the YA, YP, MA and MP respectively. Fifteen proteins were significantly differentially expressed between YA and YP, and 11 between MA and MP. Carbonic anhydrase III, lambda crystallin, alpha crystallin A and B, beta crystallin B1 and B2 were more abundant in the anterior region, whereas vimentin was less abundant in the anterior region. For comparisons between age groups, 4 proteins were differentially expressed in both YA relative to MA and YP relative to MP. Western blotting confirmed the differential expression of carbonic anhydrase III, alpha crystallin B and beta crystallin B2. The protein profiles of the vitreous humor showed age- and compartment-related differences. This differential protein profile provides a baseline for understanding the vitreous compartmentalization in the rabbit and suggests that further studies profiling proteins in different compartments of the vitreous in other species may be warranted.

No MeSH data available.


Venn diagram comparing the proteomes of four human vitreous studies (present study versus Yee et al. versus Aretz et al. Versus versus Semba et al. (made with online venn diagram plotter at http://bioinfogp.cnb.csic.es/tools/venny).
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pone.0153560.g001: Venn diagram comparing the proteomes of four human vitreous studies (present study versus Yee et al. versus Aretz et al. Versus versus Semba et al. (made with online venn diagram plotter at http://bioinfogp.cnb.csic.es/tools/venny).

Mentions: The total number of non-redundant proteins identified by LC-MS/MS in our study was 466, based on the criteria that only proteins identified with at least 2 peptides in at least 2 samples of one individual group were selected with identity confidence of 90% and higher. A previous review performed by Semba et al. reported a total of 545 non-redundant proteins in human VH [18]. Other studies such as those performed by Aretz et al.[19] and Murphy et al.[20] identified 1111 and 1205 proteins from human vitreous respectively, the latter included proteins identified with 1 unique peptide. Recently Yee et al detected 1217 proteins in fetal and young adult human vitreous, and identified differences between embryonic and young adult vitreous proteomes[21]. In this study, we identified only 466 proteins, 299 of these are new and have not been previously reported in either of aforementioned studies excluding that by Murphy et al[20] (Fig 1). Full lists of all identified proteins and newly identified proteins in our study were provided in the supplemental tables (S1 Table).


Differential Expression of Vitreous Proteins in Young and Mature New Zealand White Rabbits.

Liu Y, Bouhenni RA, Dufresne CP, Semba RD, Edward DP - PLoS ONE (2016)

Venn diagram comparing the proteomes of four human vitreous studies (present study versus Yee et al. versus Aretz et al. Versus versus Semba et al. (made with online venn diagram plotter at http://bioinfogp.cnb.csic.es/tools/venny).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835093&req=5

pone.0153560.g001: Venn diagram comparing the proteomes of four human vitreous studies (present study versus Yee et al. versus Aretz et al. Versus versus Semba et al. (made with online venn diagram plotter at http://bioinfogp.cnb.csic.es/tools/venny).
Mentions: The total number of non-redundant proteins identified by LC-MS/MS in our study was 466, based on the criteria that only proteins identified with at least 2 peptides in at least 2 samples of one individual group were selected with identity confidence of 90% and higher. A previous review performed by Semba et al. reported a total of 545 non-redundant proteins in human VH [18]. Other studies such as those performed by Aretz et al.[19] and Murphy et al.[20] identified 1111 and 1205 proteins from human vitreous respectively, the latter included proteins identified with 1 unique peptide. Recently Yee et al detected 1217 proteins in fetal and young adult human vitreous, and identified differences between embryonic and young adult vitreous proteomes[21]. In this study, we identified only 466 proteins, 299 of these are new and have not been previously reported in either of aforementioned studies excluding that by Murphy et al[20] (Fig 1). Full lists of all identified proteins and newly identified proteins in our study were provided in the supplemental tables (S1 Table).

Bottom Line: Our results showed that 231, 375, 273 and 353 proteins were identified in the YA, YP, MA and MP respectively.Fifteen proteins were significantly differentially expressed between YA and YP, and 11 between MA and MP.Western blotting confirmed the differential expression of carbonic anhydrase III, alpha crystallin B and beta crystallin B2.

View Article: PubMed Central - PubMed

Affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, United States of America.

ABSTRACT
Different anatomical regions have been defined in the vitreous humor including central vitreous, basal vitreous, vitreous cortex, vitreoretinal interface and zonule. In this study we sought to characterize changes in the proteome of vitreous humor (VH) related to compartments or age in New Zealand white rabbits (NZW). Vitreous humor was cryo-collected from young and mature New Zealand white rabbit eyes, and dissected into anterior and posterior compartments. All samples were divided into 4 groups: Young Anterior (YA), Young Posterior (YP), Mature Anterior (MA) and Mature Posterior (MP) vitreous. Tryptic digests of total proteins were analyzed by liquid chromatography followed by tandem mass spectrometry. Spectral count was used to determine the relative protein abundances and identify proteins with statistical differences between compartment and age groups. Western blotting was performed to validate some of the differentially expressed proteins. Our results showed that 231, 375, 273 and 353 proteins were identified in the YA, YP, MA and MP respectively. Fifteen proteins were significantly differentially expressed between YA and YP, and 11 between MA and MP. Carbonic anhydrase III, lambda crystallin, alpha crystallin A and B, beta crystallin B1 and B2 were more abundant in the anterior region, whereas vimentin was less abundant in the anterior region. For comparisons between age groups, 4 proteins were differentially expressed in both YA relative to MA and YP relative to MP. Western blotting confirmed the differential expression of carbonic anhydrase III, alpha crystallin B and beta crystallin B2. The protein profiles of the vitreous humor showed age- and compartment-related differences. This differential protein profile provides a baseline for understanding the vitreous compartmentalization in the rabbit and suggests that further studies profiling proteins in different compartments of the vitreous in other species may be warranted.

No MeSH data available.