Urea transport in initial IMCD of rats fed a low-protein diet: functional properties and mRNA abundance.
Feeding rats a low-protein (8%) diet (LPD) for 2 wk induces a facilitated urea transporter in rat initial inner medullary collecting ducts (IMCDs). To determine whether this is preceded by an increase in mRNA abundance, we designed degenerate polymerase chain reaction primers to the rabbit facilitated urea transporter (UT2; G. You, C. P. Smith, Y. Kanai, W.-S. Lee, M. Stelzner, and M. A. Hediger. Nature Lond. 365: 844-847, 1993) and amplified a 716-bp cDNA fragment to perform Northern analysis of the base or tip of rat inner medulla. In the base, the predominant transcript was a 2.9-kb band, which increased 55% after 1 wk on an LPD; there was no change in a 4-kb band. In the tip, the 4-kb band predominated, but neither band varied with an LPD. Next, we functionally characterized the induced urea transporter using microperfused initial IMCDs from rats fed an LPD for 2 wk. First, 100 pM arginine vasopressin (AVP) stimulated urea permeability (Purea); 10 nM AVP increased Purea further. Second, raising perfusate and bath osmolality to 690 mosmol/kgH2O (NaCl added) stimulated Purea; adding AVP (10 nM) increased Purea further. Third, thiourea reversibly inhibited AVP-stimulated Purea.(ABSTRACT TRUNCATED AT 250 WORDS)
Ashkar
Z M
ZM
Department of Medicine, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Martial
S
S
Isozaki
T
T
Price
S R
SR
Sands
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000494:pharmacology; D001483:Base Sequence; D001692:Biological Transport; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000378:metabolism; D018753:Diet, Protein-Restricted; D015870:Gene Expression / Q000187:drug effects; D066298:In Vitro Techniques; D007679:Kidney Medulla / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D007685:Kidney Tubules, Collecting / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000378:metabolism; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D013890:Thiourea / Q000494:pharmacology; D014508:Urea / Q000378:metabolism*; D000096982:Urea Transporters
D001120:Arginine; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D013890:Thiourea; D014508:Urea; D000096982:Urea Transporters