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A novel role for visual arrestin in endocytosis-mediated retinal degeneration.

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Call No.	QP609.P555 O73 2004
Author	Orem, Nicholas R., Jr.
Title	A novel role for visual arrestin in endocytosis-mediated retinal degeneration.
Date	2004
Abstract	Mutations in the Drosophila retinal-specific Phospholipase C (NORPA) cause massive retinal degeneration within five days of constant light exposure. We have previously shown that in norpA mutants persistent complexes between the light receptor (rhodopsin) and the regulatory protein (arrestin) are formed when the flies are exposed to light. We have also demonstrated that these complexes are a critical factor in norpA -mediated retinal degeneration. Furthermore, it has been shown that inhibiting apoptosis or receptor-mediated endocytosis blocks norpA -induced retinal degeneration. In order to better understand the role endocytosis plays in norpA -mediated retinal degeneration, we have performed a comprehensive study of arrestin and rhodopsin localization under various light treatments and genetic backgrounds. We now demonstrate using biochemistry and immunohistochemistry that the rhodopsin-arrestin complexes are quickly removed by endocytosis from the rhabdomere to cytoplasmic puncta in light treated norpA flies. This internalization results in at least 80% of the rhodopsin within 24 hours, which in turn leads to the internalized rhodopsin being found in cytoplasmic puncta at extremely high local concentration. Additionally, we can now show that the AP2 adaptor complex plays a central role in norpA -mediated retinal degeneration. Interestingly, both Drosophila arrestin 2 and mammalian visual arrestin contain an AP2 binding motif. We demonstrate that the AP2 binding domain of arrestin is necessary not only for norpA -mediated retinal degeneration, but also for long term photoreceptor viability as well. Furthermore, we demonstrate that the endocytosis accessory protein EPS-15 is a novel retinal degeneration mutant, and endophilin, another endocytic protein is not required for rhodopsin endocytosis or rhodopsin/arrestin complex-mediated retinal degeneration.
Description	x, 162 leaves : col. ill. 28 cm.
Format	application/pdf
Advisor	Patrick Dolph
Subject	Phospholipase C -- Mechanism of action. Retinal degeneration -- Pathophysiology. Retinal degeneration -- Genetic aspects. Endocytosis. Rhodopsin. Cellular signal transduction. Light -- Physiological effect. Arrestin. Clathrin. Adaptor Proteins
College	Dartmouth College -- Department of Biological Sciences.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Copyright	All rights reserved. No part of this book may be reproduced in any form by any electronic or mechanical means without permission in writing from the author.
DOI	http://dx.doi.org/10.1349/ddlp.41

Title	Page 1
Format	application/pdf
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Full Text	A Novel Role for Visual Arrestin in Endocytosis-mediated Retinal Degeneration A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biological Sciences by Nicholas R. Orem DARTMOUTH COLLEGE Hanover, New Hampshire November 17, 2004 Examining committee Charles K. Barlowe, Ph.D. Dean of Graduate Studies (chair) Patrick Dolph, Ph.D. Sharon Bickel, Ph.D. Thomas Jack, Ph.D. Joseph O’Tousa, Ph.D.

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