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
N2 - 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.
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.
U2 - 10.1021/acs.nanolett.8b03721
DO - 10.1021/acs.nanolett.8b03721
M3 - Journal article
C2 - 30584808
SN - 1530-6984
VL - 19
SP - 699
EP - 707
JO - Nano Letters
JF - Nano Letters
IS - 2
ER -