Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors

The dynamic organization of signaling cascades inside primary cilia is key to signal propagation. Yet little is known about the dynamics of ciliary membrane proteins besides a possible role for motor-driven Intraflagellar Transport (IFT). To characterize these dynamics, we imaged single molecules of...

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Main Authors:	Fan Ye, David K Breslow, Elena F Koslover, Andrew J Spakowitz, W James Nelson, Maxence V Nachury
Format:	Article
Language:	English
Published:	eLife Sciences Publications Ltd 2013-08-01
Series:	eLife
Subjects:	cilia IFT diffusion signaling motor
Online Access:	https://elifesciences.org/articles/00654

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spelling	doaj-b80410ab2f0741ad817a1ee410b6b6e02021-05-04T22:30:20ZengeLife Sciences Publications LtdeLife2050-084X2013-08-01210.7554/eLife.00654Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptorsFan Ye0David K Breslow1Elena F Koslover2Andrew J Spakowitz3W James Nelson4Maxence V Nachury5Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States; Department of Biology, Stanford University, Stanford, United StatesDepartment of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United StatesDepartment of Chemical Engineering, Stanford University, Stanford, United StatesDepartment of Chemical Engineering, Stanford University, Stanford, United StatesDepartment of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States; Department of Biology, Stanford University, Stanford, United StatesDepartment of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United StatesThe dynamic organization of signaling cascades inside primary cilia is key to signal propagation. Yet little is known about the dynamics of ciliary membrane proteins besides a possible role for motor-driven Intraflagellar Transport (IFT). To characterize these dynamics, we imaged single molecules of Somatostatin Receptor 3 (SSTR3, a GPCR) and Smoothened (Smo, a Hedgehog signal transducer) in the ciliary membrane. While IFT trains moved processively from one end of the cilium to the other, single SSTR3 and Smo underwent mostly diffusive behavior interspersed with short periods of directional movements. Statistical subtraction of instant velocities revealed that SSTR3 and Smo spent less than a third of their time undergoing active transport. Finally, SSTR3 and IFT movements could be uncoupled by perturbing either membrane protein diffusion or active transport. Thus ciliary membrane proteins move predominantly by diffusion, and attachment to IFT trains is transient and stochastic rather than processive or spatially determined.https://elifesciences.org/articles/00654ciliaIFTdiffusionsignalingmotor
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Fan Ye David K Breslow Elena F Koslover Andrew J Spakowitz W James Nelson Maxence V Nachury
spellingShingle	Fan Ye David K Breslow Elena F Koslover Andrew J Spakowitz W James Nelson Maxence V Nachury Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors eLife cilia IFT diffusion signaling motor
author_facet	Fan Ye David K Breslow Elena F Koslover Andrew J Spakowitz W James Nelson Maxence V Nachury
author_sort	Fan Ye
title	Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
title_short	Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
title_full	Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
title_fullStr	Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
title_full_unstemmed	Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
title_sort	single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
publisher	eLife Sciences Publications Ltd
series	eLife
issn	2050-084X
publishDate	2013-08-01
description	The dynamic organization of signaling cascades inside primary cilia is key to signal propagation. Yet little is known about the dynamics of ciliary membrane proteins besides a possible role for motor-driven Intraflagellar Transport (IFT). To characterize these dynamics, we imaged single molecules of Somatostatin Receptor 3 (SSTR3, a GPCR) and Smoothened (Smo, a Hedgehog signal transducer) in the ciliary membrane. While IFT trains moved processively from one end of the cilium to the other, single SSTR3 and Smo underwent mostly diffusive behavior interspersed with short periods of directional movements. Statistical subtraction of instant velocities revealed that SSTR3 and Smo spent less than a third of their time undergoing active transport. Finally, SSTR3 and IFT movements could be uncoupled by perturbing either membrane protein diffusion or active transport. Thus ciliary membrane proteins move predominantly by diffusion, and attachment to IFT trains is transient and stochastic rather than processive or spatially determined.
topic	cilia IFT diffusion signaling motor
url	https://elifesciences.org/articles/00654
work_keys_str_mv	AT fanye singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors AT davidkbreslow singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors AT elenafkoslover singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors AT andrewjspakowitz singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors AT wjamesnelson singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors AT maxencevnachury singlemoleculeimagingrevealsamajorrolefordiffusionintheexplorationofciliaryspacebysignalingreceptors
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Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors

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