title: Intercapillary bridging cells: immunocytochemical characteristics of cells that connect blood vessels in the retina.
abstract: Intervascular bridges are fibrous strands that connect neighboring capillaries. These strands present associated cells, intervascular bridging cells (IBCs), whose nature and functional significance remains controversial. The aim of this study was to characterize the immunophenotype of IBCs, and contribute to understand their mechanical and intercellular communication properties in the retina. Quantification and retinal distribution of IBCs were also determined. For this purpose, C57BL/6N and nestin-GFP transgenic mice, as well as human retinas, were used. Whole-mount retinas were studied by means of immunohistochemistry and cytochemistry, and isolation of retinal vasculature was achieved by trypsin/pepsin digest technique. PAS reaction and the immunolabeling with anti-collagen IV and laminin antibodies revealed that IBCs were completely surrounded by a basement membrane, connecting two or more neighboring capillaries. IBCs were scarce and their number decreased with age. They were preferentially localized in the deep vascular plexus. In a murine model of experimental glaucoma, methylcellulose injected eyes showed retinal neovascularization and increased number of IBCs in the deep vascular plexus. IBCs were marked with anti-NG2, anti-PDGFR-β and anti-CD34 antibodies, and with tomato lectin, and were negative for PECAM-1. IBCs expressed nestin and filamentous actin, but desmin and α-smooth muscle actin were not detected. Moreover, these cells expressed the gap junction protein connexin 43. These results showed that IBCs had a pericytic nature since they expressed NG2 and the receptor for PDGF-B, and they were negative for PECAM-1. However, they were marked with CD34 and the tomato lectin, suggesting that they constitute a special subtype of pericytes, sharing characteristics with endothelial cells. IBCs presumably present mechanical functions due to the presence of filamentous actin. Connexin 43 was found in IBCs, suggesting that these cells allow intercellular communication between adjacent capillaries. This may represent an advantage for vasomotor tone integration and coordination in blood vessels without innervation, such as those of the retina.
authors.affiliation: CBATEG, Autonomous University of Barcelona, E-08193 Bellaterra, Spain. lmjorge@fmv.utl.pt
authors.forename: Luísa
authors.initials: L
authors.lastname: Mendes-Jorge
authors.forename: Cristina
authors.initials: C
authors.lastname: Llombart
authors.forename: David
authors.initials: D
authors.lastname: Ramos
authors.forename: Mariana
authors.initials: M
authors.lastname: López-Luppo
authors.forename: Andreia
authors.initials: A
authors.lastname: Valença
authors.forename: Victor
authors.initials: V
authors.lastname: Nacher
authors.forename: Marc
authors.initials: M
authors.lastname: Navarro
authors.forename: Ana
authors.initials: A
authors.lastname: Carretero
authors.forename: Simón
authors.initials: S
authors.lastname: Méndez-Ferrer
authors.forename: Alfonso
authors.initials: A
authors.lastname: Rodriguez-Baeza
authors.forename: Jesús
authors.initials: J
authors.lastname: Ruberte
mesh_terms: D000328:Adult; D000369:Aged, 80 and over; D000818:Animals; D000941:Antigens / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002196:Capillaries / Q000166:cytology; D002450:Cell Communication / Q000502:physiology*; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005901:Glaucoma / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016130:Immunophenotyping; D007381:Intermediate Filament Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000235:genetics; D064231:Nestin; D020286:Pericytes / Q000166:cytology* / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism; D015861:Retinal Neovascularization / Q000378:metabolism / Q000473:pathology; D012171:Retinal Vessels / Q000166:cytology*; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D000941:Antigens; D018952:Antigens, CD34; D015415:Biomarkers; D049452:Green Fluorescent Proteins; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011134:Polysaccharides; D011509:Proteoglycans; D020797:Receptor, Platelet-Derived Growth Factor beta; D000099216:Chondroitin Sulfate Proteoglycan 4; C577484:NES protein, human; C577485:Nes protein, mouse; C037199:poly-N-acetyllactosamine