Cellular mechanisms underlying prostaglandin-induced transient cAMP signals near the plasma membrane of HEK-293 cells
| Autor: | Xuan Le, Céline Méhats, Leslie A. Piggott, Thomas C. Rich, Marco Conti, Kathryn A. Hassell, Jeffrey W. Karpen, Wenkuan Xin |
|---|---|
| Přispěvatelé: | Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston (UTHealth), Department of Pharmacology, University of South Alabama-College of Medicine and Center for Lung Biology, Department of Obstetrics and Gynecology [Stanford], Stanford Medicine, Stanford University-Stanford University, Department of Physiology and Pharmacology, Oregon Health and Science University [Portland] (OHSU), Mehats, Celine |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
MESH: Signal Transduction
Phosphodiesterase Inhibitors Physiology MESH: Sulfonamides MESH: Phosphodiesterase Inhibitors [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology Ion Channels chemistry.chemical_compound 0302 clinical medicine MESH: Rolipram Cyclic AMP MESH: Protein Kinase Inhibitors Prostaglandin E1 MESH: Cyclic AMP Sulfonamides 0303 health sciences MESH: Kinetics MESH: Peptides Phosphodiesterase Cell biology MESH: Nucleotides Cyclic Biochemistry MESH: 3' 5'-Cyclic-AMP Phosphodiesterases Nucleotides Cyclic Signal transduction Ion Channel Gating Rolipram Adenylyl Cyclases Signal Transduction MESH: Cyclic Nucleotide Phosphodiesterases Type 4 G protein Prostaglandin MESH: Cyclic AMP-Dependent Protein Kinases Buffers Biology Models Biological Article Cell Line 03 medical and health sciences MESH: Computer Simulation MESH: Adenylate Cyclase MESH: Isoquinolines Humans Computer Simulation Protein kinase A Protein Kinase Inhibitors Adaptor Proteins Signal Transducing 030304 developmental biology MESH: Adaptor Proteins Signal Transducing MESH: Humans Cell Membrane HEK 293 cells MESH: Models Biological Cell Biology MESH: Ion Channel Gating Isoquinolines Cyclic AMP-Dependent Protein Kinases [SDV.MHEP.UN] Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology Cyclic Nucleotide Phosphodiesterases Type 4 MESH: Cell Line Kinetics chemistry 3' 5'-Cyclic-AMP Phosphodiesterases Cell culture MESH: Prostaglandins MESH: Ion Channels Prostaglandins MESH: Buffers Peptides 030217 neurology & neurosurgery MESH: Cell Membrane |
| Zdroj: | American Journal of Physiology-Cell Physiology American Journal of Physiology-Cell Physiology, American Physiological Society, 2007, 292 (1), pp.C319-31. ⟨10.1152/ajpcell.00121.2006⟩ |
| ISSN: | 0363-6143 1522-1563 |
| DOI: | 10.1152/ajpcell.00121.2006⟩ |
| Popis: | We have previously used cyclic nucleotide-gated (CNG) channels as sensors to measure cAMP signals in human embryonic kidney (HEK)-293 cells. We found that prostaglandin E1(PGE1) triggered transient increases in cAMP concentration near the plasma membrane, whereas total cAMP levels rose to a steady plateau over the same time course. In addition, we presented evidence that the decline in the near-membrane cAMP levels was due primarily to a PGE1-induced stimulation of phosphodiesterase (PDE) activity, and that the differences between near-membrane and total cAMP levels were largely due to diffusional barriers and differential PDE activity. Here, we examine the mechanisms regulating transient, near-membrane cAMP signals. We observed that 5-min stimulation of HEK-293 cells with prostaglandins triggered a two- to threefold increase in PDE4 activity. Extracellular application of H89 (a PKA inhibitor) inhibited stimulation of PDE4 activity. Similarly, when we used CNG channels to monitor cAMP signals we found that both extracellular and intracellular (via the whole-cell patch pipette) application of H89, or the highly selective PKA inhibitor, PKI, prevented the decline in prostaglandin-induced responses. Following pretreatment with rolipram (a PDE4 inhibitor), H89 had little or no effect on near-membrane or total cAMP levels. Furthermore, disrupting the subcellular localization of PKA with the A-kinase anchoring protein (AKAP) disruptor Ht31 prevented the decline in the transient response. Based on these data we developed a plausible kinetic model that describes prostaglandin-induced cAMP signals. This model has allowed us to quantitatively demonstrate the importance of PKA-mediated stimulation of PDE4 activity in shaping near-membrane cAMP signals. |
| Databáze: | OpenAIRE |
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