Aquaporin-3 and aquaporin-4 are sorted differently and separately in the trans-Golgi network

Eva Arnspang Christensen, S. Sundbye, W. J. Nelson, L. N. Nejsum

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Aquaporin-3 (AQP3) and aquaporin-4 (AQP4) are homologous proteins expressed in the basolateral plasma membrane of kidney collecting duct principal cells, where they mediate the exit pathway for apically reabsorbed water. Although both proteins are localized to the same plasma membrane domain, it is unknown if they are sorted together in the Golgi, or arrive in the same or different vesicles at the plasma membrane. We addressed these questions using high resolution deconvolution imaging, spinning disk and laser scanning confocal microscopy of cells expressing AQP3 and AQP4. AQP3 and AQP4 were observed mostly in separate post-Golgi carriers, and spinning disk microscopy showed that most of AQP3 and AQP4 were delivered to the plasma membrane in separate vesicles. In contrast, VSV-G and LDL-R, two well-characterized basolateral proteins, co-localized to a high degree in the same post-Golgi carriers, indicating that the differential sorting of AQP3 and AQP4 is specific and regulated. Significantly, a chimeric AQP3 containing the AQP4 cytoplasmic tails co-localized with AQP4 in post-Golgi vesicles. These results indicate that AQP3 and AQP4 are separated into different post-Golgi carriers based on different cytoplasmic domain sorting signals, and are then delivered separately to the plasma membrane.
Original languageEnglish
JournalPLOS ONE
Volume8
Pages (from-to)e73977
ISSN1932-6203
DOIs
Publication statusPublished - 2013

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Aquaporin 3
Aquaporin 4
trans-Golgi Network
aquaporins
Cell membranes
Cell Membrane
plasma membrane
Sorting
Collecting Kidney Tubules
Proteins
Confocal microscopy
spinning
Deconvolution
sorting
Confocal Microscopy
Ducts
Microscopy
Microscopic examination
Cells
Scanning

Keywords

  • Animals Aquaporin 3/genetics/*metabolism Aquaporin 4/genetics/*metabolism Dogs Gene Expression Genes, Reporter Green Fluorescent Proteins/genetics/metabolism Madin Darby Canine Kidney Cells Microscopy/methods Molecular Imaging Mutant Chimeric Proteins/genetics/*metabolism Protein Transport Recombinant Fusion Proteins/genetics/*metabolism Transport Vesicles/*metabolism Water/metabolism trans-Golgi Network/*metabolism

Cite this

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title = "Aquaporin-3 and aquaporin-4 are sorted differently and separately in the trans-Golgi network",
abstract = "Aquaporin-3 (AQP3) and aquaporin-4 (AQP4) are homologous proteins expressed in the basolateral plasma membrane of kidney collecting duct principal cells, where they mediate the exit pathway for apically reabsorbed water. Although both proteins are localized to the same plasma membrane domain, it is unknown if they are sorted together in the Golgi, or arrive in the same or different vesicles at the plasma membrane. We addressed these questions using high resolution deconvolution imaging, spinning disk and laser scanning confocal microscopy of cells expressing AQP3 and AQP4. AQP3 and AQP4 were observed mostly in separate post-Golgi carriers, and spinning disk microscopy showed that most of AQP3 and AQP4 were delivered to the plasma membrane in separate vesicles. In contrast, VSV-G and LDL-R, two well-characterized basolateral proteins, co-localized to a high degree in the same post-Golgi carriers, indicating that the differential sorting of AQP3 and AQP4 is specific and regulated. Significantly, a chimeric AQP3 containing the AQP4 cytoplasmic tails co-localized with AQP4 in post-Golgi vesicles. These results indicate that AQP3 and AQP4 are separated into different post-Golgi carriers based on different cytoplasmic domain sorting signals, and are then delivered separately to the plasma membrane.",
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Aquaporin-3 and aquaporin-4 are sorted differently and separately in the trans-Golgi network. / Christensen, Eva Arnspang; Sundbye, S.; Nelson, W. J.; Nejsum, L. N.

In: PLOS ONE, Vol. 8, 2013, p. e73977.

Research output: Contribution to journalJournal articleResearchpeer-review

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T1 - Aquaporin-3 and aquaporin-4 are sorted differently and separately in the trans-Golgi network

AU - Christensen, Eva Arnspang

AU - Sundbye, S.

AU - Nelson, W. J.

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AB - Aquaporin-3 (AQP3) and aquaporin-4 (AQP4) are homologous proteins expressed in the basolateral plasma membrane of kidney collecting duct principal cells, where they mediate the exit pathway for apically reabsorbed water. Although both proteins are localized to the same plasma membrane domain, it is unknown if they are sorted together in the Golgi, or arrive in the same or different vesicles at the plasma membrane. We addressed these questions using high resolution deconvolution imaging, spinning disk and laser scanning confocal microscopy of cells expressing AQP3 and AQP4. AQP3 and AQP4 were observed mostly in separate post-Golgi carriers, and spinning disk microscopy showed that most of AQP3 and AQP4 were delivered to the plasma membrane in separate vesicles. In contrast, VSV-G and LDL-R, two well-characterized basolateral proteins, co-localized to a high degree in the same post-Golgi carriers, indicating that the differential sorting of AQP3 and AQP4 is specific and regulated. Significantly, a chimeric AQP3 containing the AQP4 cytoplasmic tails co-localized with AQP4 in post-Golgi vesicles. These results indicate that AQP3 and AQP4 are separated into different post-Golgi carriers based on different cytoplasmic domain sorting signals, and are then delivered separately to the plasma membrane.

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