Dynein and dynactin colocalize with AQP2 water channels in intracellular vesicles from kidney collecting duct.
We investigated whether the motor protein cytoplasmic dynein and dynactin, a protein complex thought to link dynein with vesicles, are present in rat renal collecting ducts and associated with aquaporin-2 (AQP2)-bearing vesicles. Immunoblotting demonstrated cytoplasmic dynein heavy and intermediate chains in kidney, with relative expression levels of inner medulla > outer medulla > cortex. In addition to being present in cytoplasmic fractions, dynein was abundant in membrane fractions enriched for intracellular vesicles. Dynactin was also abundant in membrane fractions enriched for intracellular vesicles. Furthermore, both dynactin and dynein were present in vesicles specifically immunoisolated using anti-AQP2 antibodies. Immunocytochemistry revealed labeling for dynein in the collecting duct principal cells with a pattern consistent with labeling of intracellular vesicles. Moreover, quantitative double immunogold labeling confirmed colocalization of AQP2 and dynein in the same vesicles at the electron microscopic level. Thus the microtubule-associated motor protein dynein and the associated dynactin complex are present in rat renal collecting duct principal cells and are associated with intracellular vesicles, including those bearing AQP2, consistent with the view that dynein and dynactin are involved in vasopressin-regulated trafficking of AQP2-bearing vesicles.
Marples
D
D
Department of Physiology, University of Leeds, United Kingdom.
Schroer
T A
TA
Ahrens
N
N
Taylor
A
A
Knepper
M A
MA
Nielsen
S
S
D000818:Animals; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D002462:Cell Membrane / Q000737:chemistry; D000072159:Dynactin Complex; D004398:Dyneins / Q000032:analysis*; D015151:Immunoblotting; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000737:chemistry; D007473:Ion Channels / Q000032:analysis*; D007679:Kidney Medulla / Q000737:chemistry; D007685:Kidney Tubules, Collecting / Q000737:chemistry* / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar
C496025:Aqp2 protein, rat; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D000072159:Dynactin Complex; D007473:Ion Channels; D008869:Microtubule-Associated Proteins; D004398:Dyneins