Localization of aquaporin-7 in rat and mouse kidney using RT-PCR, immunoblotting, and immunocytochemistry.
To establish the segmental, cellular, and subcellular localization of AQP7 in rat and mouse kidney, we used RT-PCR, immunocytochemical, and immunoblotting approaches. RT-PCR of rat and mouse kidney zones revealed AQP7 mRNA in cortex and outer stripe of the outer medulla. RT-PCR on microdissected nephron segments revealed AQP7 mRNA in proximal convoluted and straight tubules. Immunoblotting using peptide-derived rabbit antibodies to either rat or mouse AQP7 revealed a 28-kDa band in kidney and testes from rat and mouse, respectively. Immunocytochemistry revealed strong AQP7 labeling of segment 3 proximal tubules and weaker labeling of proximal convoluted tubules in both rat and mouse kidneys. The labeling was almost exclusively confined to the brush border with no basolateral labeling. No labeling was observed of thin descending limbs or collecting duct. Immunolabeling controls were negative. The presence of AQP7 in the proximal tubule brush border indicates a role of AQP7 in proximal tubule water reabsorption.
Nejsum
L N
LN
Department of Cell Biology, Institute of Anatomy, Aarhus, DK-8000, Denmark.
Elkjaer
M
M
Hager
H
H
Frokiaer
J
J
Kwon
T H
TH
Nielsen
S
S
D000818:Animals; D000906:Antibodies / Q000276:immunology; D020346:Aquaporins; D015139:Blotting, Southern; D015153:Blotting, Western; D007150:Immunohistochemistry; D007473:Ion Channels / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D007668:Kidney / Q000737:chemistry*; D007687:Kidney Tubules, Proximal / Q000737:chemistry; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000906:Antibodies; C491927:Aqp7 protein, mouse; C491926:Aqp7 protein, rat; D020346:Aquaporins; D007473:Ion Channels; D012333:RNA, Messenger