Phagocytosis of rod outer segments by retinal pigment epithelial cells requires alpha(v)beta5 integrin for binding but not for internalization.
Phagocytosis of shed photoreceptor rod outer segments (ROS) by the retinal pigment epithelium (RPE) is essential for retinal function. Here, we demonstrate that this process requires alpha(v)beta5 integrin, rather than alpha(v)beta3 integrin utilized by systemic macrophages. Although adult rat RPE expressed both alpha(v)beta3 and alpha(v)beta5 integrins, only alpha(v)beta3 was expressed at birth, when the retina is immature and phagocytosis is absent. Expression of alpha(v)beta5 was first detected in RPE at PN7 and reached adult levels at PN11, just before onset of phagocytic activity. Interestingly, alpha(v)beta5 localized in vivo to the apical plasma membrane, facing the photoreceptors, and to intracellular vesicles, whereas alpha(v)beta3 was expressed basolaterally. Using quantitative fluorimaging to assess in vitro uptake of fluorescent particles by human (ARPE-19) and rat (RPE-J) cell lines, alpha(v)beta5 function-blocking antibodies were shown to reduce phagocytosis by drastically decreasing (85%) binding of ROS but not of latex beads. In agreement with a role for alpha(v)beta5 in phagocytosis, immunofluorescence experiments demonstrated codistribution of alpha(v)beta5 integrin with internalized ROS. Control experiments showed that blocking alpha(v)beta3 function with antibodies did not inhibit ROS phagocytosis and that alpha(v)beta3 did not colocalize with phagocytosed ROS. Taken together, our results indicate that the RPE requires the integrin receptor alpha(v)beta5 specifically for the binding of ROS and that phagocytosis involves internalization of a ROS-alpha(v)beta5 complex. Alpha(v)beta5 integrin does not participate in phagocytosis by other phagocytic cells and is the first of the RPE receptors involved in ROS phagocytosis that may be specific for this process.
Finnemann
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SC
Margaret M. Dyson Vision Research Institute, Department of Ophthalmology, Cornell University Medical College, New York, NY 10021, USA.
Bonilha
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VL
Marmorstein
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AD
Rodriguez-Boulan
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E
D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D016023:Integrins / Q000378:metabolism*; D010587:Phagocytosis; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000276:immunology*; D011485:Protein Binding; D019030:Receptors, Vitronectin; D012374:Rod Cell Outer Segment / Q000276:immunology*
D016023:Integrins; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5