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Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors
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| Zeitschriftentitel: | American Journal of Physiology-Renal Physiology |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | American Journal of Physiology-Renal Physiology, 287, 2004, 2, S. F204-F214 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Physiological Society
|
| Schlagwörter: |
| author_facet |
Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. |
|---|---|
| author |
Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. |
| spellingShingle |
Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. American Journal of Physiology-Renal Physiology Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors Physiology |
| author_sort |
xia, shen-ling |
| spelling |
Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. 1931-857X 1522-1466 American Physiological Society Physiology http://dx.doi.org/10.1152/ajprenal.00281.2003 <jats:p>Kidney tubules are targets for the activation of locally released nucleotides through multiple P2 receptor types. Activation of these P2 receptors modulates cellular Ca<jats:sup>2+</jats:sup>signaling and downstream cellular function. The purpose of this study was to determine whether P2 receptors were present in mIMCD-3 cells, a mouse inner medullary collecting duct cell line, and if so, to examine their link with intracellular Ca<jats:sup>2+</jats:sup>homeostasis. To monitor intracellular Ca<jats:sup>2+</jats:sup>concentration ([Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>), experiments were conducted using the fluorescent dye fura 2. ATP (0.1–100 μM) produced a dose-dependent increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in a physiological Ca<jats:sup>2+</jats:sup>-containing solution, with an EC<jats:sub>50</jats:sub>of 2.5 μM. The P2-receptor antagonist PPADS reduced the effect of ATP on [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, and the P1-receptor agonist adenosine caused only a small increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>. Preincubation of cells with the phospholipase C antagonist U-73122 blocked the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, indicating P2Y receptors were involved in this process. In a Ca<jats:sup>2+</jats:sup>-free bath solution, thapsigargin and ATP induced intracellular Ca<jats:sup>2+</jats:sup>release from an identical pool. Nucleotides caused an increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in the potency order of UTP = ATP > ATPγS > ADP > UDP that is best fitted with the P2Y<jats:sub>2</jats:sub>subtype profile. Although the P2Y agonist UTP induced a similar large transient increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>as did ATP, a small but sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>occurred only in ATP-stimulated cells, suggesting the role of P2X receptors in Ca<jats:sup>2+</jats:sup>influx. The sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>could be blocked by either nonselective cation channel blockers Gd<jats:sup>3+</jats:sup>or P2X antagonists PPADS and PPNDS. Furthermore, when either Gd<jats:sup>3+</jats:sup>or PPNDS was applied to the bath solution before ATP application, the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>was significantly reduced. Both RT-PCR and Western blotting corroborated the presence of P2X<jats:sub>1</jats:sub>and P2Y<jats:sub>2</jats:sub>receptors. These studies demonstrate that mIMCD-3 cells have both P2X and P2Y subtype receptors and that the activation of both P2X and P2Y receptors by extracellular ATP appears to be required to regulate intracellular Ca<jats:sup>2+</jats:sup>signaling.</jats:p> Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors American Journal of Physiology-Renal Physiology |
| doi_str_mv |
10.1152/ajprenal.00281.2003 |
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Online Free |
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Biologie |
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ElectronicArticle |
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ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTE1Mi9hanByZW5hbC4wMDI4MS4yMDAz |
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American Physiological Society, 2004 |
| imprint_str_mv |
American Physiological Society, 2004 |
| issn |
1931-857X 1522-1466 |
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1931-857X 1522-1466 |
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English |
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American Physiological Society (CrossRef) |
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xia2004extracellularatpinducedcalciumsignalinginmimcd3cellsrequiresbothp2xandp2ypurinoceptors |
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2004 |
| publisher |
American Physiological Society |
| recordtype |
ai |
| record_format |
ai |
| series |
American Journal of Physiology-Renal Physiology |
| source_id |
49 |
| title |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_unstemmed |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_full |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_fullStr |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_full_unstemmed |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_short |
Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_sort |
extracellular atp-induced calcium signaling in mimcd-3 cells requires both p2x and p2y purinoceptors |
| topic |
Physiology |
| url |
http://dx.doi.org/10.1152/ajprenal.00281.2003 |
| publishDate |
2004 |
| physical |
F204-F214 |
| description |
<jats:p>Kidney tubules are targets for the activation of locally released nucleotides through multiple P2 receptor types. Activation of these P2 receptors modulates cellular Ca<jats:sup>2+</jats:sup>signaling and downstream cellular function. The purpose of this study was to determine whether P2 receptors were present in mIMCD-3 cells, a mouse inner medullary collecting duct cell line, and if so, to examine their link with intracellular Ca<jats:sup>2+</jats:sup>homeostasis. To monitor intracellular Ca<jats:sup>2+</jats:sup>concentration ([Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>), experiments were conducted using the fluorescent dye fura 2. ATP (0.1–100 μM) produced a dose-dependent increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in a physiological Ca<jats:sup>2+</jats:sup>-containing solution, with an EC<jats:sub>50</jats:sub>of 2.5 μM. The P2-receptor antagonist PPADS reduced the effect of ATP on [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, and the P1-receptor agonist adenosine caused only a small increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>. Preincubation of cells with the phospholipase C antagonist U-73122 blocked the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, indicating P2Y receptors were involved in this process. In a Ca<jats:sup>2+</jats:sup>-free bath solution, thapsigargin and ATP induced intracellular Ca<jats:sup>2+</jats:sup>release from an identical pool. Nucleotides caused an increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in the potency order of UTP = ATP > ATPγS > ADP > UDP that is best fitted with the P2Y<jats:sub>2</jats:sub>subtype profile. Although the P2Y agonist UTP induced a similar large transient increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>as did ATP, a small but sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>occurred only in ATP-stimulated cells, suggesting the role of P2X receptors in Ca<jats:sup>2+</jats:sup>influx. The sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>could be blocked by either nonselective cation channel blockers Gd<jats:sup>3+</jats:sup>or P2X antagonists PPADS and PPNDS. Furthermore, when either Gd<jats:sup>3+</jats:sup>or PPNDS was applied to the bath solution before ATP application, the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>was significantly reduced. Both RT-PCR and Western blotting corroborated the presence of P2X<jats:sub>1</jats:sub>and P2Y<jats:sub>2</jats:sub>receptors. These studies demonstrate that mIMCD-3 cells have both P2X and P2Y subtype receptors and that the activation of both P2X and P2Y receptors by extracellular ATP appears to be required to regulate intracellular Ca<jats:sup>2+</jats:sup>signaling.</jats:p> |
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| description | <jats:p>Kidney tubules are targets for the activation of locally released nucleotides through multiple P2 receptor types. Activation of these P2 receptors modulates cellular Ca<jats:sup>2+</jats:sup>signaling and downstream cellular function. The purpose of this study was to determine whether P2 receptors were present in mIMCD-3 cells, a mouse inner medullary collecting duct cell line, and if so, to examine their link with intracellular Ca<jats:sup>2+</jats:sup>homeostasis. To monitor intracellular Ca<jats:sup>2+</jats:sup>concentration ([Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>), experiments were conducted using the fluorescent dye fura 2. ATP (0.1–100 μM) produced a dose-dependent increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in a physiological Ca<jats:sup>2+</jats:sup>-containing solution, with an EC<jats:sub>50</jats:sub>of 2.5 μM. The P2-receptor antagonist PPADS reduced the effect of ATP on [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, and the P1-receptor agonist adenosine caused only a small increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>. Preincubation of cells with the phospholipase C antagonist U-73122 blocked the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, indicating P2Y receptors were involved in this process. In a Ca<jats:sup>2+</jats:sup>-free bath solution, thapsigargin and ATP induced intracellular Ca<jats:sup>2+</jats:sup>release from an identical pool. Nucleotides caused an increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in the potency order of UTP = ATP > ATPγS > ADP > UDP that is best fitted with the P2Y<jats:sub>2</jats:sub>subtype profile. Although the P2Y agonist UTP induced a similar large transient increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>as did ATP, a small but sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>occurred only in ATP-stimulated cells, suggesting the role of P2X receptors in Ca<jats:sup>2+</jats:sup>influx. The sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>could be blocked by either nonselective cation channel blockers Gd<jats:sup>3+</jats:sup>or P2X antagonists PPADS and PPNDS. Furthermore, when either Gd<jats:sup>3+</jats:sup>or PPNDS was applied to the bath solution before ATP application, the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>was significantly reduced. Both RT-PCR and Western blotting corroborated the presence of P2X<jats:sub>1</jats:sub>and P2Y<jats:sub>2</jats:sub>receptors. These studies demonstrate that mIMCD-3 cells have both P2X and P2Y subtype receptors and that the activation of both P2X and P2Y receptors by extracellular ATP appears to be required to regulate intracellular Ca<jats:sup>2+</jats:sup>signaling.</jats:p> |
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| imprint | American Physiological Society, 2004 |
| imprint_str_mv | American Physiological Society, 2004 |
| institution | DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4 |
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| physical | F204-F214 |
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| publisher | American Physiological Society |
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| series | American Journal of Physiology-Renal Physiology |
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| spelling | Xia, Shen-Ling Wang, Lanjun Cash, Melanie N. Teng, Xueling Schwalbe, Ruth A. Wingo, Charles S. 1931-857X 1522-1466 American Physiological Society Physiology http://dx.doi.org/10.1152/ajprenal.00281.2003 <jats:p>Kidney tubules are targets for the activation of locally released nucleotides through multiple P2 receptor types. Activation of these P2 receptors modulates cellular Ca<jats:sup>2+</jats:sup>signaling and downstream cellular function. The purpose of this study was to determine whether P2 receptors were present in mIMCD-3 cells, a mouse inner medullary collecting duct cell line, and if so, to examine their link with intracellular Ca<jats:sup>2+</jats:sup>homeostasis. To monitor intracellular Ca<jats:sup>2+</jats:sup>concentration ([Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>), experiments were conducted using the fluorescent dye fura 2. ATP (0.1–100 μM) produced a dose-dependent increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in a physiological Ca<jats:sup>2+</jats:sup>-containing solution, with an EC<jats:sub>50</jats:sub>of 2.5 μM. The P2-receptor antagonist PPADS reduced the effect of ATP on [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, and the P1-receptor agonist adenosine caused only a small increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>. Preincubation of cells with the phospholipase C antagonist U-73122 blocked the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>, indicating P2Y receptors were involved in this process. In a Ca<jats:sup>2+</jats:sup>-free bath solution, thapsigargin and ATP induced intracellular Ca<jats:sup>2+</jats:sup>release from an identical pool. Nucleotides caused an increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>in the potency order of UTP = ATP > ATPγS > ADP > UDP that is best fitted with the P2Y<jats:sub>2</jats:sub>subtype profile. Although the P2Y agonist UTP induced a similar large transient increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>as did ATP, a small but sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>occurred only in ATP-stimulated cells, suggesting the role of P2X receptors in Ca<jats:sup>2+</jats:sup>influx. The sustained increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>could be blocked by either nonselective cation channel blockers Gd<jats:sup>3+</jats:sup>or P2X antagonists PPADS and PPNDS. Furthermore, when either Gd<jats:sup>3+</jats:sup>or PPNDS was applied to the bath solution before ATP application, the ATP-induced increase in [Ca<jats:sup>2+</jats:sup>]<jats:sub>i</jats:sub>was significantly reduced. Both RT-PCR and Western blotting corroborated the presence of P2X<jats:sub>1</jats:sub>and P2Y<jats:sub>2</jats:sub>receptors. These studies demonstrate that mIMCD-3 cells have both P2X and P2Y subtype receptors and that the activation of both P2X and P2Y receptors by extracellular ATP appears to be required to regulate intracellular Ca<jats:sup>2+</jats:sup>signaling.</jats:p> Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors American Journal of Physiology-Renal Physiology |
| spellingShingle | Xia, Shen-Ling, Wang, Lanjun, Cash, Melanie N., Teng, Xueling, Schwalbe, Ruth A., Wingo, Charles S., American Journal of Physiology-Renal Physiology, Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors, Physiology |
| title | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_full | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_fullStr | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_full_unstemmed | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_short | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| title_sort | extracellular atp-induced calcium signaling in mimcd-3 cells requires both p2x and p2y purinoceptors |
| title_unstemmed | Extracellular ATP-induced calcium signaling in mIMCD-3 cells requires both P2X and P2Y purinoceptors |
| topic | Physiology |
| url | http://dx.doi.org/10.1152/ajprenal.00281.2003 |