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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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 |
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