Identification of Type VI Collagen in the Trabecular Meshwork and Expression of its mRNA by Trabecular Cells

Brenda J Tripathi; Marlan Hansen; Junping Li; Ramesh C Tripathi

doi:10.1006/exer.1994.1006

Back

Identification of Type VI Collagen in the Trabecular Meshwork and Expression of its mRNA by Trabecular Cells

Journal article

Peer reviewed

Identification of Type VI Collagen in the Trabecular Meshwork and Expression of its mRNA by Trabecular Cells

Brenda J Tripathi, Marlan Hansen, Junping Li and Ramesh C Tripathi

Experimental eye research, Vol.58(2), pp.181-188

1994

DOI: 10.1006/exer.1994.1006

PMID: 8157110

View Online

Abstract

To investigate the nature of the 140 kDa glycoprotein in the trabecular meshwork, polypeptides were extracted with either urea/sodium dodecyl sulfate (SDS)/beta-2-mercaptoethanol (BME) or guanidine hydrochloride followed by pepsin digestion. After electrophoresis and immunoblotting with anti-type-VI-collagen antibodies, a single fraction of molecular weight 140 kDa was identified in the urea/SDS/BME extracts. Pepsin solubilization revealed two immunoreactive fractions (molecular weights 75 and 85 kDa) that comigrated with purified, pepsin-solubilized type VI collagen. By using the polymerase chain reaction (PCR) and primers specific for the α2(VI) chain of type VI collagen, a single PCR product was obtained, which corresponded to the expected size of 137 base pairs, from the total RNA extracted from the trabecular meshwork ex vivo. Southern hybridization with the antisense oligonucleotide probe of the α2(VI) chain confirmed that the amplified sequence was specific. The results show that the trabecular meshwork contains a significant amount of type VI collagen and that trabecular cells express the mRNA coding for the α2(VI) chain of this glycoprotein. The presence of type VI collagen in the trabecular meshwork is implicated in cell-extracellular matrix interactions at this site, and its abnormal accumulation in glaucomatous and aging eyes probably signifies a defect in the function of the trabecular cells in these eyes.

primary open-angle glaucoma

aqueous outflow

cell adhesion

immunoblotting

Southern hybridization

gel electrophoresis

long-spacing collagen

age-related changes

extracellular matrix

polymerase chain reaction

Details

Title: Subtitle: Identification of Type VI Collagen in the Trabecular Meshwork and Expression of its mRNA by Trabecular Cells
Creators: Brenda J Tripathi
Marlan Hansen
Junping Li
Ramesh C Tripathi
Resource Type: Journal article
Publication Details: Experimental eye research, Vol.58(2), pp.181-188
Publisher: Elsevier Ltd
DOI: 10.1006/exer.1994.1006
PMID: 8157110
ISSN: 0014-4835
eISSN: 1096-0007
Language: English
Date published: 1994
Academic Unit: Molecular Physiology and Biophysics; Neurosurgery; Otolaryngology
Record Identifier: 9984006426802771

Metrics

20 Record Views

17 Times Cited - Web of Science