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Mechanism of sphingosine 1-phosphate- and lysophosphatidic acid-induced up-regulation of adhesion molecules and eosinophil chemoattractant in nerve cells.

Costello RW, Maloney M, Atiyeh M, Gleich G, Walsh MT - Int J Mol Sci (2011)

Bottom Line: IMR-32 neuroblastoma cells were used as an in vitro cholinergic nerve cell model.The G(i) coupled receptors S1P(1), S1P(3), LPA(1), LPA(2) and LPA(3) were expressed on IMR-32 cells.Both S1P and LPA induced ERK phosphorylation and ERK- and G(i)-dependent up-regulation of ICAM-1 expression, with differing time courses.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Royal College of Surgeons in Ireland, Beaumont Hospital, Dublin 9, Ireland; E-Mails: rcostello@rcsi.ie (R.W.C.); micmaloney@rcsi.ie (M.M.); matiyeh@rcsi.ie (M.A.).

ABSTRACT
The lysophospholipids sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) act via G-protein coupled receptors S1P(1-5) and LPA(1-3) respectively, and are implicated in allergy. Eosinophils accumulate at innervating cholinergic nerves in asthma and adhere to nerve cells via intercellular adhesion molecule-1 (ICAM-1). IMR-32 neuroblastoma cells were used as an in vitro cholinergic nerve cell model. The G(i) coupled receptors S1P(1), S1P(3), LPA(1), LPA(2) and LPA(3) were expressed on IMR-32 cells. Both S1P and LPA induced ERK phosphorylation and ERK- and G(i)-dependent up-regulation of ICAM-1 expression, with differing time courses. LPA also induced ERK- and G(i)-dependent up-regulation of the eosinophil chemoattractant, CCL-26. The eosinophil granule protein eosinophil peroxidase (EPO) induced ERK-dependent up-regulation of transcription of S1P(1), LPA(1), LPA(2) and LPA(3), providing the situation whereby eosinophil granule proteins may enhance S1P- and/or LPA- induced eosinophil accumulation at nerve cells in allergic conditions.

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Eosinophil peroxidase induces ERK-dependent transcriptional up-regulation of LPA1, LPA2 and LPA3 in IMR32 cells. IMR32 cells in differentiation medium were pre-treated or not overnight with PD98059 (50 M). Cells were then treated with eosinophil peroxidase (EPO) (1 μg/mL) for the indicated times then harvested for RNA and cDNA preparation and real-time PCR using primers for: (A) LPA1 or (B) LPA2 or (C) LPA3 or β-actin (A, B and C). Results are expressed as EPO-induced fold increase in LPA receptor/β-actin ratio over non-EPO treated cells harvested at the same time as the 24 h time point, which are set to unity. Inserts show baseline absolute values of LPA receptor versus β-actin in non-EPO treated cells, cultured in the presence or absence of PD98059 pre-treatment, and harvested at the same time as the 24 hour time point cells. Mean ± SEM; * p < 0.05, *** p < 0.001, EPO-induced fold increase in LPA receptor over untreated; † p < 0.05, ††† p < 0.001, PD98059-induced reduction in EPO-mediated S1P1 up-regulation.
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f4-ijms-12-03237: Eosinophil peroxidase induces ERK-dependent transcriptional up-regulation of LPA1, LPA2 and LPA3 in IMR32 cells. IMR32 cells in differentiation medium were pre-treated or not overnight with PD98059 (50 M). Cells were then treated with eosinophil peroxidase (EPO) (1 μg/mL) for the indicated times then harvested for RNA and cDNA preparation and real-time PCR using primers for: (A) LPA1 or (B) LPA2 or (C) LPA3 or β-actin (A, B and C). Results are expressed as EPO-induced fold increase in LPA receptor/β-actin ratio over non-EPO treated cells harvested at the same time as the 24 h time point, which are set to unity. Inserts show baseline absolute values of LPA receptor versus β-actin in non-EPO treated cells, cultured in the presence or absence of PD98059 pre-treatment, and harvested at the same time as the 24 hour time point cells. Mean ± SEM; * p < 0.05, *** p < 0.001, EPO-induced fold increase in LPA receptor over untreated; † p < 0.05, ††† p < 0.001, PD98059-induced reduction in EPO-mediated S1P1 up-regulation.

Mentions: We have previously shown that eosinophil adhesion to IMR32 cells induces eosinophil degranulation and release of granule proteins, including eosinophil peroxidase (EPO) [8–10]. To determine whether released eosinophil granule protein could increase expression of IMR32 S1P or LPA receptors, nerve cells were treated with eosinophil peroxidase (EPO) (1 μg/mL) for 1, 4, 18 or 24 h in the presence or absence of PD98059. Real-time PCR was used to monitor the expression of S1P1 and S1P3 receptors (Figure 3) or LPA1, LPA2 and LPA3 receptors (Figure 4). EPO significantly enhanced expression of S1P1 by approximately 3–4 fold between 4–18 h of stimulation (Figure 3A). This up-regulation was dependent on ERK activation as it was abolished in the presence of PD98059. S1P3 expression was unaffected by EPO (Figure 3B). LPA1 (Figure 4A), LPA2 (Figure 4B) and LPA3 (Figure 4C) expression were all raised approximately 3-fold at 18 h of stimulation, then fell back to control levels by 24 h.


Mechanism of sphingosine 1-phosphate- and lysophosphatidic acid-induced up-regulation of adhesion molecules and eosinophil chemoattractant in nerve cells.

Costello RW, Maloney M, Atiyeh M, Gleich G, Walsh MT - Int J Mol Sci (2011)

Eosinophil peroxidase induces ERK-dependent transcriptional up-regulation of LPA1, LPA2 and LPA3 in IMR32 cells. IMR32 cells in differentiation medium were pre-treated or not overnight with PD98059 (50 M). Cells were then treated with eosinophil peroxidase (EPO) (1 μg/mL) for the indicated times then harvested for RNA and cDNA preparation and real-time PCR using primers for: (A) LPA1 or (B) LPA2 or (C) LPA3 or β-actin (A, B and C). Results are expressed as EPO-induced fold increase in LPA receptor/β-actin ratio over non-EPO treated cells harvested at the same time as the 24 h time point, which are set to unity. Inserts show baseline absolute values of LPA receptor versus β-actin in non-EPO treated cells, cultured in the presence or absence of PD98059 pre-treatment, and harvested at the same time as the 24 hour time point cells. Mean ± SEM; * p < 0.05, *** p < 0.001, EPO-induced fold increase in LPA receptor over untreated; † p < 0.05, ††† p < 0.001, PD98059-induced reduction in EPO-mediated S1P1 up-regulation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3116188&req=5

f4-ijms-12-03237: Eosinophil peroxidase induces ERK-dependent transcriptional up-regulation of LPA1, LPA2 and LPA3 in IMR32 cells. IMR32 cells in differentiation medium were pre-treated or not overnight with PD98059 (50 M). Cells were then treated with eosinophil peroxidase (EPO) (1 μg/mL) for the indicated times then harvested for RNA and cDNA preparation and real-time PCR using primers for: (A) LPA1 or (B) LPA2 or (C) LPA3 or β-actin (A, B and C). Results are expressed as EPO-induced fold increase in LPA receptor/β-actin ratio over non-EPO treated cells harvested at the same time as the 24 h time point, which are set to unity. Inserts show baseline absolute values of LPA receptor versus β-actin in non-EPO treated cells, cultured in the presence or absence of PD98059 pre-treatment, and harvested at the same time as the 24 hour time point cells. Mean ± SEM; * p < 0.05, *** p < 0.001, EPO-induced fold increase in LPA receptor over untreated; † p < 0.05, ††† p < 0.001, PD98059-induced reduction in EPO-mediated S1P1 up-regulation.
Mentions: We have previously shown that eosinophil adhesion to IMR32 cells induces eosinophil degranulation and release of granule proteins, including eosinophil peroxidase (EPO) [8–10]. To determine whether released eosinophil granule protein could increase expression of IMR32 S1P or LPA receptors, nerve cells were treated with eosinophil peroxidase (EPO) (1 μg/mL) for 1, 4, 18 or 24 h in the presence or absence of PD98059. Real-time PCR was used to monitor the expression of S1P1 and S1P3 receptors (Figure 3) or LPA1, LPA2 and LPA3 receptors (Figure 4). EPO significantly enhanced expression of S1P1 by approximately 3–4 fold between 4–18 h of stimulation (Figure 3A). This up-regulation was dependent on ERK activation as it was abolished in the presence of PD98059. S1P3 expression was unaffected by EPO (Figure 3B). LPA1 (Figure 4A), LPA2 (Figure 4B) and LPA3 (Figure 4C) expression were all raised approximately 3-fold at 18 h of stimulation, then fell back to control levels by 24 h.

Bottom Line: IMR-32 neuroblastoma cells were used as an in vitro cholinergic nerve cell model.The G(i) coupled receptors S1P(1), S1P(3), LPA(1), LPA(2) and LPA(3) were expressed on IMR-32 cells.Both S1P and LPA induced ERK phosphorylation and ERK- and G(i)-dependent up-regulation of ICAM-1 expression, with differing time courses.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Royal College of Surgeons in Ireland, Beaumont Hospital, Dublin 9, Ireland; E-Mails: rcostello@rcsi.ie (R.W.C.); micmaloney@rcsi.ie (M.M.); matiyeh@rcsi.ie (M.A.).

ABSTRACT
The lysophospholipids sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) act via G-protein coupled receptors S1P(1-5) and LPA(1-3) respectively, and are implicated in allergy. Eosinophils accumulate at innervating cholinergic nerves in asthma and adhere to nerve cells via intercellular adhesion molecule-1 (ICAM-1). IMR-32 neuroblastoma cells were used as an in vitro cholinergic nerve cell model. The G(i) coupled receptors S1P(1), S1P(3), LPA(1), LPA(2) and LPA(3) were expressed on IMR-32 cells. Both S1P and LPA induced ERK phosphorylation and ERK- and G(i)-dependent up-regulation of ICAM-1 expression, with differing time courses. LPA also induced ERK- and G(i)-dependent up-regulation of the eosinophil chemoattractant, CCL-26. The eosinophil granule protein eosinophil peroxidase (EPO) induced ERK-dependent up-regulation of transcription of S1P(1), LPA(1), LPA(2) and LPA(3), providing the situation whereby eosinophil granule proteins may enhance S1P- and/or LPA- induced eosinophil accumulation at nerve cells in allergic conditions.

Show MeSH
Related in: MedlinePlus