Stimulation by vasopressin of ERK phosphorylation and vector-driven water flux in astrocytes is transactivation-dependent.
Vasopressin acts on astrocytic Gq protein- and phospholipase C-coupled V1 receptors. In mesangial cells, which also express the V1 receptor, it stimulates cell growth by activating mitogen-activated protein kinase (MAP kinase) secondary to transactivation of the epidermal growth factor (EGF) receptor. Transactivation is an intracellular/extracellular process, in which activation of a Gq or a Gi/o protein-coupled receptor leads to metalloproteinase-catalyzed shedding of an EGF receptor agonist, which stimulates EGF receptors on the same cell and/or its neighbor(s). The goal of the present study was to investigate if vasopressin signaling is mediated by transactivation also in astrocytes and whether such a transactivation is required for its ability to facilitate vector-driven water fluxes. Vasopressin concentrations between 10(-12) and 10(-6) M were found to lead to phosphorylation (activation) of extracellular regulated kinase 1 and 2 (ERK 1/2). Phosphorylation of ERK 1/2 could be completely inhibited by either AG1478, an inhibitor of the EGF receptor-activated tyrosine kinase, or GM6001, an inhibitor of Zn2+-activated metalloproteinases, indicating the involvement of transactivation. Exposure to a hypotonic medium caused an immediate (within one min) increase in cell water volume (demonstrated by decrease of fluorescence quenching of calcein), part of which was dependent upon the presence of vasopressin, added at a concentration of 1 x 10(-8) M. This vasopressin-dependent component persisted throughout the duration of the experiment (22 min). The effect of vasopressin was abolished in the presence of AG1478, indicating its dependence upon transactivation, and by U0126 an inhibitor of the MAP kinase/ERK kinase (MEK), and thus of ERK 1/2 phosphorylation.
Du
Ting
T
Department of Clinical Pharmacology, China Medical University, PR China.
Song
Dan
D
Li
Hongmei
H
Li
Baoman
B
Cai
Liping
L
Hertz
Leif
L
Peng
Liang
L
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D002070:Butadienes / Q000494:pharmacology; D048429:Cell Size; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D009570:Nitriles / Q000494:pharmacology; D010766:Phosphorylation; D011799:Quinazolines; D013268:Stimulation, Chemical; D015533:Transcriptional Activation / Q000187:drug effects / Q000502:physiology*; D020032:Tyrphostins / Q000494:pharmacology; D014667:Vasopressins / Q000494:pharmacology*; D014867:Water / Q000378:metabolism*
D002070:Butadienes; D004791:Enzyme Inhibitors; D009570:Nitriles; D011799:Quinazolines; D020032:Tyrphostins; C113580:U 0126; D014867:Water; D014667:Vasopressins; C101044:RTKI cpd; D048049:Extracellular Signal-Regulated MAP Kinases; D020928:Mitogen-Activated Protein Kinases