Cytoskeleton-dependent endocytosis is required for apical type 1 angiotensin II receptor-mediated phospholipase C activation in cultured rat proximal tubule cells.
Renal proximal tubule sodium reabsorption is enhanced by apical or basolateral angiotensin II (AII). Although AII activates phospholipase C (PLC) in other tissues, AII coupling to PLC on either apical or basolateral surfaces of proximal tubule cells is unclear. To determine if AII causes PLC activation, and the differences between apical and basolateral AII receptor function, receptors were unilaterally activated in rat proximal tubule cells cultured on permeable, collagen-coated supports. Apical AII incubation resulted in concentration- and time-dependent inositol trisphosphate (IP3) formation. Basolateral AII caused greater IP3 responses. Apical AII-induced IP3 generation was inhibited by DuP 753, suggesting that the type 1 AII receptor subtype mediated proximal tubule PLC activation. Apical AII signaling did not result from paracellular ligand leak to basolateral receptors since AII-induced PLC activation occurred when basolateral AII receptors were occupied by Sar-Leu AII or DuP 753. Inhibition of endocytosis with phenylarsine oxide prevented apical (but not basolateral) AII-induced IP3 formation. Cytoskeletal disruption with colchicine or cytochalasin D also prevented apical AII-induced IP3 generation. These results demonstrate that in cultured rat proximal tubule cells, AII is coupled to PLC via type 1 AII receptors and cytoskeleton-dependent endocytosis is required for apical (but not basolateral) AII receptor-mediated PLC activation.
Schelling
J R
JR
University of Colorado Health Sciences Center, Denver 80262.
Hanson
A S
AS
Marzec
R
R
Linas
S L
SL
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000494:pharmacology; D016764:Cell Polarity; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology; D004705:Endocytosis; D004789:Enzyme Activation; D007093:Imidazoles / Q000494:pharmacology; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D019808:Losartan; D008297:Male; D008854:Microscopy, Electron; D010281:Parathyroid Hormone / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011945:Receptors, Angiotensin / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D013777:Tetrazoles / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism*
D001713:Biphenyl Compounds; D007093:Imidazoles; D010281:Parathyroid Hormone; D011945:Receptors, Angiotensin; D013777:Tetrazoles; D000804:Angiotensin II; D001127:Arginine Vasopressin; D015638:Cytochalasin D; D015544:Inositol 1,4,5-Trisphosphate; D012964:Sodium; D000242:Cyclic AMP; D010738:Type C Phospholipases; D019808:Losartan