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Semaphorin4D-PlexinB1 Signaling Attenuates Photoreceptor Outer Segment Phagocytosis by Reducing Rac1 Activity of RPE Cells

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Abstract

Semaphorins form a family of secreted and membrane-bound molecules that were identified originally as axonal guidance factors during neuronal development. Given their wide distribution in many including mature tissues, semaphorin 4D (sema4D) and its main functional receptor plexin B1 (plxnB1) are expected to fulfill additional functions that remain to be uncovered. A main characteristic of the plexin receptor family is its ability to reorganize the cytoskeleton. PlxnB1 specifically may directly interact with Rho family GTPases to regulate F-actin driven pathways in cells in culture. Diurnal clearance phagocytosis by the retinal pigment epithelium (RPE) of photoreceptor outer segment fragments (POS) is critical for photoreceptor function and longevity. In this process, rearrangement of RPE cytoskeletal F-actin via activation of the Rho family GTPase Rac1 is essential for POS internalization. Here, we show a novel role in POS phagocytosis by RPE cells in culture and in vivo for plexin B1 and its ligand sema4D. Exogenous sema4D abolishes POS internalization (but not binding) by differentiated RPE cells in culture by decreasing the GTP load of Rac1. In the rat eye, sema4D localizes to retinal photoreceptors, while PlxnB1 is expressed by neighboring RPE cells. At the peak of diurnal retinal phagocytosis after light onset, plxnB1 phosphorylation and sema4D levels are reduced in wild-type rat retina in situ but not in mutant RCS rat retina in which the RPE lacks phagocytic activity. Finally, increased POS phagosome content after light onset is observed in the RPE in situ of mice with either plxnB1 or sema4D gene deletion. Altogether, our results demonstrate a novel physiological function for sema4D/plxnB1 signaling in RPE phagocytosis serving as attenuating brake prior to light onset whose release enables the diurnal phagocytic burst.

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Acknowledgments

This project was supported by NIH grant EY26215 (to S.C.F) and National Eye Institute intramural funds (to A.M.). We are grateful to Drs. Eszter Paldy and Rohini Kunar (University of Heidelberg, Germany) who generously provided the dissected tissues from plxnB1 KO mice for this study.

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Authors and Affiliations

  1. Department of Biological Sciences Center for Cancer, Genetic Diseases, and Gene Regulation, Fordham University, Larkin Hall, 441 East Fordham Road, Bronx, NY, 10458, USA

    Ayelen Bulloj & Silvia C. Finnemann

  2. Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Arvydas Maminishkis

  3. Department of Nephrology, Osaka University Graduate School of Medicine, Osaka, 565-0871, Japan

    Masayuki Mizui

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  1. Ayelen Bulloj
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  2. Arvydas Maminishkis
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  3. Masayuki Mizui
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  4. Silvia C. Finnemann
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Correspondence to Silvia C. Finnemann.

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All human tissue research followed the tenets of the Declaration of Helsinki and the NIH institutional review board. All procedures were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and reviewed and approved according to institutional requirements.

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The authors declare that they have no conflict of interest.

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Bulloj, A., Maminishkis, A., Mizui, M. et al. Semaphorin4D-PlexinB1 Signaling Attenuates Photoreceptor Outer Segment Phagocytosis by Reducing Rac1 Activity of RPE Cells. Mol Neurobiol 55, 4320–4332 (2018). https://doi.org/10.1007/s12035-017-0649-5

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