The role of P2Y1 purinergic receptors and cytosolic Ca2+ in hypotonically activated osmolyte efflux from a rat hepatoma cell line
| Autor: | Pauline R. Junankar, Kiaran Kirk, Ari Karjalainen |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Thapsigargin
Biochemistry chemistry.chemical_compound Receptors Purinergic P2Y1 Adenosine Triphosphate Cytosol Liver Neoplasms Experimental medicine Extracellular Purinergic P2 Receptor Antagonists Tumor Cells Cultured Animals Receptor Molecular Biology DNA Primers Base Sequence Chemistry Apyrase Receptors Purinergic P2 Reverse Transcriptase Polymerase Chain Reaction Purinergic receptor Osmolar Concentration Biological Transport Cell Biology Adenosine Molecular biology Rats Adenosine Diphosphate Adenosine diphosphate Biophysics Calcium Adenosine triphosphate medicine.drug |
| Zdroj: | The Journal of biological chemistry. 277(43) |
| ISSN: | 0021-9258 |
| Popis: | Exposure of HTC rat hepatoma cells to a 33% decrease in extracellular osmolality caused the cytosolic Ca(2+) concentration ([Ca(2+)](i)) to increase transiently by approximately 90 nm. This rise in [Ca(2+)](i) was inhibited strongly by apyrase, grade VII (which has a low ATP/ADPase ratio) but not by apyrase grade VI (which has a high ATP/ADPase ratio) or hexokinase, indicating that extracellular ADP and/or ATP play a role in the [Ca(2+)](i) increase. The hypotonically induced rise in [Ca(2+)](i) was prevented by the prior discharge of the intracellular Ca(2+) store of the cells by thapsigargin. Removal of extracellular Ca(2+) or inhibition of Ca(2+) influx by 1-10 microm Gd(3+) depleted the thapsigargin-sensitive Ca(2+) stores and thereby diminished the rise in [Ca(2+)](i). The hypotonically induced rise in [Ca(2+)](i) was prevented by adenosine 2'-phosphate-5'-phosphate (A2P5P) and pyridoxyl-5'-phosphate-6-azophenyl-2',4'-disulfonate, inhibitors of purinergic P2Y(1) receptors for which ADP is a major agonist. Both inhibitors also blocked the rise in [Ca(2+)](i) elicited by addition of ADP to cells in isotonic medium, whereas A2P5P had no effect on the rise in [Ca(2+)](i) elicited by the addition of the P2Y(2) and P2Y(4) receptor agonist, UTP. HTC cells were shown to express mRNA encoding for rat P2Y(1), P2Y(2), and P2Y(6) receptors. Inhibition of the hypotonically induced rise in [Ca(2+)](i) blocked hypotonically induced K(+) ((86)Rb(+)) efflux, modulated the hypotonically induced efflux of taurine, but had no significant effect on Cl(-) ((125)I-) efflux. The interaction of extracellular ATP and/or ADP with P2Y(1) purinergic receptors therefore plays a role in the response of HTC cells to osmotic swelling but does not account for activation of all the efflux pathways involved in the volume-regulatory response. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|