Parathyroid hormone (PTH)/PTH-related peptide receptor density modulates activation of phospholipase C and phosphate transport by PTH in LLC-PK1 cells.
We showed previously that a single species of cloned PTH/PTH-related peptide (PTHrP) receptors, when stably expressed in LLC-PK1 kidney cells, couples to multiple second messenger signals and biological responses. To address the linkages of individual messenger signals to specific biological responses in these cells, we examined the relations among PTH/PTHrP receptor expression, PTH-activated phospholipase C (PLC) and adenylyl cyclase, and PTH-regulated phosphate transport in LLC-PK1 cells that stably express cloned rat PTH/PTHrP receptors. Among 18 such subclones, PTH stimulation of intracellular cAMP accumulation was nearly equivalent, despite differences in receptor density ranging from 20,000-400,000 sites/cell. In contrast, activation of PLC by PTH was directly and continuously dependent upon receptor density. PTH-stimulated phosphate uptake also was strongly dependent upon receptor expression, correlated well with PLC activity, was mimicked by active phorbol esters but not by cAMP analogs or forskolin, and was strikingly inhibited by the protein kinase C inhibitor, staurosporine. The peptide analog [Arg2]human PTH-(1-34), which significantly stimulated cAMP accumulation but failed to activate PLC, also did not increase phosphate uptake. We conclude that in LLC-PK1 cells, PTH-modulated PLC activation, unlike adenylyl cyclase activation, is strongly dependent upon PTH/PTHrP receptor density. This feature is reflected in the analogous relation between receptor density and PTH regulation of phosphate uptake, which appears to be mediated via a PKC-dependent pathway in these transfected cells. The results suggest that regulation of PTH/PTHrP receptor expression on target cells may provide a mechanism for altering the character as well as the magnitude of the signaling response to the hormone.
Guo
J
J
Endocrine Unit, Massachusetts General Hospital, Boston 02114, USA.
Iida-Klein
A
A
Huang
X
X
Abou-Samra
A B
AB
Segre
G V
GV
Bringhurst
F R
FR
D000262:Adenylyl Cyclases / Q000378:metabolism / Q000502:physiology; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002116:Calcitonin / Q000494:pharmacology; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D018374:LLC-PK1 Cells / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D010281:Parathyroid Hormone / Q000494:pharmacology*; D010703:Phorbol Esters / Q000494:pharmacology; D010710:Phosphates / Q000493:pharmacokinetics*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D044168:Receptor, Parathyroid Hormone, Type 1; D018016:Receptors, Parathyroid Hormone / Q000032:analysis / Q000502:physiology*; D015290:Second Messenger Systems / Q000502:physiology; D019311:Staurosporine; D013552:Swine; D010738:Type C Phospholipases / Q000378:metabolism* / Q000502:physiology
D000470:Alkaloids; D007295:Inositol Phosphates; D010281:Parathyroid Hormone; D010703:Phorbol Esters; D010710:Phosphates; D044168:Receptor, Parathyroid Hormone, Type 1; D018016:Receptors, Parathyroid Hormone; D005576:Colforsin; D002116:Calcitonin; D000242:Cyclic AMP; D011493:Protein Kinase C; D010738:Type C Phospholipases; D000262:Adenylyl Cyclases; D019311:Staurosporine