Mapping the differential distribution of proteoglycan core proteins in the adult human retina, choroid, and sclera.
OBJECTIVE
To examine the presence and distribution of proteoglycan (PG) core proteins in the adult human retina, choroid, and sclera.


METHODS
Postmortem human eye tissue was dissected into Bruch's membrane/choroid complex, isolated Bruch's membrane, or neurosensory retina. PGs were extracted and partially purified by anion exchange chromatography. Trypsinized peptides were analyzed by tandem mass spectrometry and PG core proteins identified by database search. The distribution of PGs was examined by immunofluorescence microscopy on human macular tissue sections.


RESULTS
The basement membrane PGs perlecan, agrin, and collagen-XVIII were identified in the human retina, and were present in the internal limiting membrane, blood vessel walls, and Bruch's membrane. The hyalectans versican and aggrecan were also detected. Versican was identified in Bruch's membrane, while aggrecan was distributed throughout the retina, choroid, and sclera. The cartilage link protein HAPLN1 was abundant in the interphotoreceptor matrix and sclera, while HAPLN4 (brain link protein 2) was found throughout the retina and choroid. The small leucine-rich repeat PG (SLRP) family members biglycan, decorin, fibromodulin, lumican, mimecan, opticin, and prolargin were present, with different patterns of distribution in the retina, choroid, and sclera.


CONCLUSIONS
A combination of proteomics and immunohistochemistry approaches has provided for the first time a comprehensive analysis of the presence and distribution of PG core proteins throughout the human retina, choroid, and sclera. This complements our knowledge of glycosaminoglycan chain distribution in the human eye, and has important implications for understanding the structure and functional regulation of the eye in health and disease.
Centre for Ophthalmology and Vision Research, Institute of Human Development, Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
Tiarnan D L
TD
Keenan
Simon J
SJ
Clark
Richard D
RD
Unwin
Liam A
LA
Ridge
Anthony J
AJ
Day
Paul N
PN
Bishop
D000368:Aged; D000369:Aged, 80 and over; D002102:Cadaver; D002829:Choroid / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D006801:Humans; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D011509:Proteoglycans / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D012160:Retina / Q000737:chemistry*; D012590:Sclera / Q000737:chemistry*
D011509:Proteoglycans