Yongji Wang1, Kristin Repyak, Nader Sheibani. 1. Department of Ophthalmology Visual Sciences, University of Wisconsin Medical School, Madison, WI 53792-4673, USA.
Abstract
PURPOSE: Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a cell adhesion-signaling molecule with important roles in angiogenesis and inflammation. The alternative splicing of the PECAM-1 cytoplasmic domain modulates its adhesive properties during vascular development and angiogenesis. This study was designed to identify alternatively spliced PECAM-1 isoforms in human and murine retina and during postnatal vascularization of murine retina. METHODS: RT-PCR, DNA sequencing, and Western blot analysis were utilized to examine the expression pattern of alternatively spliced PECAM-1 isoforms in human and mouse retina, and during vascularization of murine retina. RESULTS: We demonstrate that the PECAM-1 cytoplasmic domain undergoes alternative splicing generating multiple isoforms in vascular beds of human and mouse retina. We detected full length PECAM-1 and an isoform that lacks exon 14 (Delta14) in human retina. Seven isoforms of PECAM-1 were detected in murine retina. These included full length, Delta12, Delta14, Delta15, Delta1215, Delta1415, and Delta12, 1415 PECAM-1 isoforms. The full length PECAM-1 was the predominant isoform detected in human retina, while the isoform lacking exon 1415 (Delta1415) was the predominant isoform detected in murine retina. In addition, the expression pattern of PECAM-1 isoforms changed during vascularization of murine retina. CONCLUSIONS: PECAM-1 mRNA undergoes alternative splicing generating multiple isoforms in human and murine retinal vasculature. The regulated expression pattern of these isoforms may influence endothelial cell adhesive properties impacting vasculogenesis and angiogenesis.
PURPOSE:Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a cell adhesion-signaling molecule with important roles in angiogenesis and inflammation. The alternative splicing of the PECAM-1 cytoplasmic domain modulates its adhesive properties during vascular development and angiogenesis. This study was designed to identify alternatively spliced PECAM-1 isoforms in human and murine retina and during postnatal vascularization of murine retina. METHODS: RT-PCR, DNA sequencing, and Western blot analysis were utilized to examine the expression pattern of alternatively spliced PECAM-1 isoforms in human and mouse retina, and during vascularization of murine retina. RESULTS: We demonstrate that the PECAM-1 cytoplasmic domain undergoes alternative splicing generating multiple isoforms in vascular beds of human and mouse retina. We detected full length PECAM-1 and an isoform that lacks exon 14 (Delta14) in human retina. Seven isoforms of PECAM-1 were detected in murine retina. These included full length, Delta12, Delta14, Delta15, Delta1215, Delta1415, and Delta12, 1415 PECAM-1 isoforms. The full length PECAM-1 was the predominant isoform detected in human retina, while the isoform lacking exon 1415 (Delta1415) was the predominant isoform detected in murine retina. In addition, the expression pattern of PECAM-1 isoforms changed during vascularization of murine retina. CONCLUSIONS:PECAM-1 mRNA undergoes alternative splicing generating multiple isoforms in human and murine retinal vasculature. The regulated expression pattern of these isoforms may influence endothelial cell adhesive properties impacting vasculogenesis and angiogenesis.
Authors: SunYoung Park; Terri A DiMaio; Elizabeth A Scheef; Christine M Sorenson; Nader Sheibani Journal: Am J Physiol Cell Physiol Date: 2010-09-01 Impact factor: 4.249