Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy

Eva C. Arnspang, Prabuddha Sengupta, Kim Mortensen, Helene Halkjær Jensen, Ute Hahn, Eva B. Vedel Jensen, Jennifer Lippincott-Schwartz, Lene Niemann Nejsum

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Several aquaporin (AQP) water channels are short-term regulated by the messenger cyclic adenosine monophosphate (cAMP), including AQP3. Bulk measurements show that cAMP can change diffusive properties of AQP3; however, it remains unknown how elevated cAMP affects AQP3 organization at the nanoscale. Here we analyzed AQP3 nano-organization following cAMP stimulation using photoactivated localization microscopy (PALM) of fixed cells combined with pair correlation analysis. Moreover, in live cells, we combined PALM acquisitions of single fluorophores with single-particle tracking (spt-PALM). These analyses revealed that AQP3 tends to cluster and that the diffusive mobility is confined to nanodomains with radii of ∼150 nm. This domain size increases by ∼30% upon elevation of cAMP, which, however, is not accompanied by a significant increase in the confined diffusion coefficient. This regulation of AQP3 organization at the nanoscale may be important for understanding the mechanisms of water AQP3-mediated water transport across plasma membranes.
Original languageEnglish
JournalNano Letters
Volume19
Issue number2
Pages (from-to)699–707
ISSN1530-6984
DOIs
Publication statusPublished - 2019

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Aquaporin 3
cyclic AMP
Aquaporins
Cell membranes
Cyclic AMP
Microscopic examination
membranes
microscopy
water
Water
Fluorophores
cells
stimulation
acquisition
diffusion coefficient
radii

Cite this

Arnspang, Eva C. ; Sengupta, Prabuddha ; Mortensen, Kim ; Jensen, Helene Halkjær ; Hahn, Ute ; Jensen, Eva B. Vedel ; Lippincott-Schwartz, Jennifer ; Nejsum, Lene Niemann. / Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy. In: Nano Letters. 2019 ; Vol. 19, No. 2. pp. 699–707.
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title = "Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy",
abstract = "Several aquaporin (AQP) water channels are short-term regulated by the messenger cyclic adenosine monophosphate (cAMP), including AQP3. Bulk measurements show that cAMP can change diffusive properties of AQP3; however, it remains unknown how elevated cAMP affects AQP3 organization at the nanoscale. Here we analyzed AQP3 nano-organization following cAMP stimulation using photoactivated localization microscopy (PALM) of fixed cells combined with pair correlation analysis. Moreover, in live cells, we combined PALM acquisitions of single fluorophores with single-particle tracking (spt-PALM). These analyses revealed that AQP3 tends to cluster and that the diffusive mobility is confined to nanodomains with radii of ∼150 nm. This domain size increases by ∼30{\%} upon elevation of cAMP, which, however, is not accompanied by a significant increase in the confined diffusion coefficient. This regulation of AQP3 organization at the nanoscale may be important for understanding the mechanisms of water AQP3-mediated water transport across plasma membranes.",
author = "Arnspang, {Eva C.} and Prabuddha Sengupta and Kim Mortensen and Jensen, {Helene Halkj{\ae}r} and Ute Hahn and Jensen, {Eva B. Vedel} and Jennifer Lippincott-Schwartz and Nejsum, {Lene Niemann}",
year = "2019",
doi = "10.1021/acs.nanolett.8b03721",
language = "English",
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Arnspang, EC, Sengupta, P, Mortensen, K, Jensen, HH, Hahn, U, Jensen, EBV, Lippincott-Schwartz, J & Nejsum, LN 2019, 'Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy', Nano Letters, vol. 19, no. 2, pp. 699–707. https://doi.org/10.1021/acs.nanolett.8b03721

Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy. / Arnspang, Eva C.; Sengupta, Prabuddha ; Mortensen, Kim; Jensen, Helene Halkjær; Hahn, Ute ; Jensen, Eva B. Vedel; Lippincott-Schwartz, Jennifer ; Nejsum, Lene Niemann.

In: Nano Letters, Vol. 19, No. 2, 2019, p. 699–707.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Regulation of Plasma Membrane Nano-Domains of the Water Channel Aquaporin-3 Revealed by Fixed and Live Photoactivated Localization Microscopy

AU - Arnspang, Eva C.

AU - Sengupta, Prabuddha

AU - Mortensen, Kim

AU - Jensen, Helene Halkjær

AU - Hahn, Ute

AU - Jensen, Eva B. Vedel

AU - Lippincott-Schwartz, Jennifer

AU - Nejsum, Lene Niemann

PY - 2019

Y1 - 2019

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AB - Several aquaporin (AQP) water channels are short-term regulated by the messenger cyclic adenosine monophosphate (cAMP), including AQP3. Bulk measurements show that cAMP can change diffusive properties of AQP3; however, it remains unknown how elevated cAMP affects AQP3 organization at the nanoscale. Here we analyzed AQP3 nano-organization following cAMP stimulation using photoactivated localization microscopy (PALM) of fixed cells combined with pair correlation analysis. Moreover, in live cells, we combined PALM acquisitions of single fluorophores with single-particle tracking (spt-PALM). These analyses revealed that AQP3 tends to cluster and that the diffusive mobility is confined to nanodomains with radii of ∼150 nm. This domain size increases by ∼30% upon elevation of cAMP, which, however, is not accompanied by a significant increase in the confined diffusion coefficient. This regulation of AQP3 organization at the nanoscale may be important for understanding the mechanisms of water AQP3-mediated water transport across plasma membranes.

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DO - 10.1021/acs.nanolett.8b03721

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VL - 19

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JO - Nano Letters

JF - Nano Letters

SN - 1530-6984

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