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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 of proteins identified in anterior or posterior vitreous of young and mature rabbits by LC-MS/MS.(A) 231 proteins were identified in the anterior vitreous of young rabbit (YA) while 375 proteins were identified in the posterior vitreous of young rabbit (YP), among them were 204 proteins shared by these two groups; (B) The total numbers of proteins detected in the anterior and posterior vitreous of mature rabbit (MA and MP respectively) were 273 and 353, they shared 238 proteins; (C) The number of common proteins shared by YA and MA was 194; (D) 295 proteins were identified in both YP and MP.
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pone.0153560.g002: Venn diagram of proteins identified in anterior or posterior vitreous of young and mature rabbits by LC-MS/MS.(A) 231 proteins were identified in the anterior vitreous of young rabbit (YA) while 375 proteins were identified in the posterior vitreous of young rabbit (YP), among them were 204 proteins shared by these two groups; (B) The total numbers of proteins detected in the anterior and posterior vitreous of mature rabbit (MA and MP respectively) were 273 and 353, they shared 238 proteins; (C) The number of common proteins shared by YA and MA was 194; (D) 295 proteins were identified in both YP and MP.

Mentions: Furthermore, as shown in Fig 2A, a total of 402 proteins were identified in young rabbits. Among them, 231 proteins were identified in young anterior vitreous while 375 proteins in young posterior vitreous. These two groups shared 204 proteins. Similarly, a total of 388 proteins were identified in the mature rabbits (Fig 2B). Among these, 273 and 353 proteins were identified in mature anterior and mature posterior vitreous, respectively. These two groups shared 238 proteins. When samples were grouped by region, a total of 310 proteins were identified in anterior compartment and 194 proteins were found common to young anterior and mature anterior vitreous (Fig 2C). In the posterior compartment, 433 proteins were identified and 295 proteins were common to young posterior and mature posterior vitreous (Fig 2D).


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 of proteins identified in anterior or posterior vitreous of young and mature rabbits by LC-MS/MS.(A) 231 proteins were identified in the anterior vitreous of young rabbit (YA) while 375 proteins were identified in the posterior vitreous of young rabbit (YP), among them were 204 proteins shared by these two groups; (B) The total numbers of proteins detected in the anterior and posterior vitreous of mature rabbit (MA and MP respectively) were 273 and 353, they shared 238 proteins; (C) The number of common proteins shared by YA and MA was 194; (D) 295 proteins were identified in both YP and MP.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153560.g002: Venn diagram of proteins identified in anterior or posterior vitreous of young and mature rabbits by LC-MS/MS.(A) 231 proteins were identified in the anterior vitreous of young rabbit (YA) while 375 proteins were identified in the posterior vitreous of young rabbit (YP), among them were 204 proteins shared by these two groups; (B) The total numbers of proteins detected in the anterior and posterior vitreous of mature rabbit (MA and MP respectively) were 273 and 353, they shared 238 proteins; (C) The number of common proteins shared by YA and MA was 194; (D) 295 proteins were identified in both YP and MP.
Mentions: Furthermore, as shown in Fig 2A, a total of 402 proteins were identified in young rabbits. Among them, 231 proteins were identified in young anterior vitreous while 375 proteins in young posterior vitreous. These two groups shared 204 proteins. Similarly, a total of 388 proteins were identified in the mature rabbits (Fig 2B). Among these, 273 and 353 proteins were identified in mature anterior and mature posterior vitreous, respectively. These two groups shared 238 proteins. When samples were grouped by region, a total of 310 proteins were identified in anterior compartment and 194 proteins were found common to young anterior and mature anterior vitreous (Fig 2C). In the posterior compartment, 433 proteins were identified and 295 proteins were common to young posterior and mature posterior vitreous (Fig 2D).

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.