LIF-mediated maintenance of PACAP-induced neurite outgrowths in human SH-SY5Y neuroblastoma cells require PI3K but not STAT or ERK signaling pathways

Eve M. Lutz, Thomas K. Monaghan, Chantevy Pou, Robin Plevin, Christopher J. MacKenzie

Research output: Contribution to journalConference Contribution

Abstract

Previously, we have shown that pituitary adenylate cyclase-activating polypeptide (PACAP) induces neuritogenesis in human SH-SY5Y cells through a cAMP-dependent but protein kinase A-independent mechanism involving activation of the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways. However, the morphological changes induced by PACAP are transient, with the increase in number of neurite-bearing cells peaking at day 4 before returning to control cell levels over an 8-day time course. Control cells maintain an undifferentiated neuroblast morphology during the 8-day experiment. On the other hand, neurite extensions are maintained by cells co-treated with PACAP and the neuropoietic cytokine leukemia inhibitory factor (LIF), with similar levels in the number of neurite-bearing cells observed at day 8 to that at day 4. Interestingly, the induction of neurite outgrowths in SH-SY5Y cells requires a minimal 2-h treatment with PACAP, with 4–6 h of treatment inducing similar peak levels of neurite-bearing cells observed at day 4 to that of continual PACAP treatment. LIF is not required until day 4 after which continual treatment with LIF is necessary for neurite maintenance in PACAP-treated cells. The signaling pathways activated by LIF were investigated also. LIF elicits signal transducer and activator of transcription 3 (STAT3) but not STAT1 tyrosine phosphorylation along with a transient activation of the ERK and prolonged activation of phosphatidylinositol-3 kinase (PI3K)/protein kinase B (PKB/Akt) in PACAP-treated and control SH-SY5Y cells. Phosphorylation of serine 727 on STAT3 was detected in PACAP-treated but not control cells, and this was prevented by the MEK-1 inhibitor PD98059. Activation of p38 MAPK or c-jun N-terminal kinase (JNK) were not detected, however, an elevated basal level of phosphorylated p38 MAP kinase was observed in PACAP-treated cells. Neither PD98059 nor siRNA knockdown of STAT3 prevented LIF-mediated maintenance of PACAP-induced neurite extensions. However, the PI3K inhibitor wortmannin was found to effectively block LIF maintenance of neurite outgrowths in PACAP-treated cells. Thus, the ability of LIF to stabilize and maintain PACAP-induced neurite outgrowths requires PI3K but not the STAT3 or ERK signaling pathways in SH-SY5Y cells.
LanguageEnglish
Pages297-297
Number of pages1
JournalJournal of Molecular Neuroscience
Volume42
Issue number3
DOIs
Publication statusPublished - Nov 2010

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Phosphatidylinositol 3-Kinase
Pituitary Adenylate Cyclase-Activating Polypeptide
Leukemia Inhibitory Factor
Extracellular Signal-Regulated MAP Kinases
Neuroblastoma
Maintenance
Neurites
STAT3 Transcription Factor
p38 Mitogen-Activated Protein Kinases
Cyclic AMP-Dependent Protein Kinases
Neuronal Outgrowth
Phosphorylation
Proto-Oncogene Proteins c-akt
JNK Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinase Kinases

Keywords

  • neurite
  • neuroblastoma cells
  • signaling pathways

Cite this

@article{012d5059597c4eaba993b3759aa0a315,
title = "LIF-mediated maintenance of PACAP-induced neurite outgrowths in human SH-SY5Y neuroblastoma cells require PI3K but not STAT or ERK signaling pathways",
abstract = "Previously, we have shown that pituitary adenylate cyclase-activating polypeptide (PACAP) induces neuritogenesis in human SH-SY5Y cells through a cAMP-dependent but protein kinase A-independent mechanism involving activation of the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways. However, the morphological changes induced by PACAP are transient, with the increase in number of neurite-bearing cells peaking at day 4 before returning to control cell levels over an 8-day time course. Control cells maintain an undifferentiated neuroblast morphology during the 8-day experiment. On the other hand, neurite extensions are maintained by cells co-treated with PACAP and the neuropoietic cytokine leukemia inhibitory factor (LIF), with similar levels in the number of neurite-bearing cells observed at day 8 to that at day 4. Interestingly, the induction of neurite outgrowths in SH-SY5Y cells requires a minimal 2-h treatment with PACAP, with 4–6 h of treatment inducing similar peak levels of neurite-bearing cells observed at day 4 to that of continual PACAP treatment. LIF is not required until day 4 after which continual treatment with LIF is necessary for neurite maintenance in PACAP-treated cells. The signaling pathways activated by LIF were investigated also. LIF elicits signal transducer and activator of transcription 3 (STAT3) but not STAT1 tyrosine phosphorylation along with a transient activation of the ERK and prolonged activation of phosphatidylinositol-3 kinase (PI3K)/protein kinase B (PKB/Akt) in PACAP-treated and control SH-SY5Y cells. Phosphorylation of serine 727 on STAT3 was detected in PACAP-treated but not control cells, and this was prevented by the MEK-1 inhibitor PD98059. Activation of p38 MAPK or c-jun N-terminal kinase (JNK) were not detected, however, an elevated basal level of phosphorylated p38 MAP kinase was observed in PACAP-treated cells. Neither PD98059 nor siRNA knockdown of STAT3 prevented LIF-mediated maintenance of PACAP-induced neurite extensions. However, the PI3K inhibitor wortmannin was found to effectively block LIF maintenance of neurite outgrowths in PACAP-treated cells. Thus, the ability of LIF to stabilize and maintain PACAP-induced neurite outgrowths requires PI3K but not the STAT3 or ERK signaling pathways in SH-SY5Y cells.",
keywords = "neurite, neuroblastoma cells, signaling pathways",
author = "Lutz, {Eve M.} and Monaghan, {Thomas K.} and Chantevy Pou and Robin Plevin and MacKenzie, {Christopher J.}",
year = "2010",
month = "11",
doi = "10.1007/s12031-009-9324-2",
language = "English",
volume = "42",
pages = "297--297",
journal = "Journal of Molecular Neuroscience",
issn = "0895-8696",
number = "3",

}

LIF-mediated maintenance of PACAP-induced neurite outgrowths in human SH-SY5Y neuroblastoma cells require PI3K but not STAT or ERK signaling pathways. / Lutz, Eve M.; Monaghan, Thomas K.; Pou, Chantevy; Plevin, Robin; MacKenzie, Christopher J.

In: Journal of Molecular Neuroscience, Vol. 42, No. 3, 11.2010, p. 297-297.

Research output: Contribution to journalConference Contribution

TY - JOUR

T1 - LIF-mediated maintenance of PACAP-induced neurite outgrowths in human SH-SY5Y neuroblastoma cells require PI3K but not STAT or ERK signaling pathways

AU - Lutz, Eve M.

AU - Monaghan, Thomas K.

AU - Pou, Chantevy

AU - Plevin, Robin

AU - MacKenzie, Christopher J.

PY - 2010/11

Y1 - 2010/11

N2 - Previously, we have shown that pituitary adenylate cyclase-activating polypeptide (PACAP) induces neuritogenesis in human SH-SY5Y cells through a cAMP-dependent but protein kinase A-independent mechanism involving activation of the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways. However, the morphological changes induced by PACAP are transient, with the increase in number of neurite-bearing cells peaking at day 4 before returning to control cell levels over an 8-day time course. Control cells maintain an undifferentiated neuroblast morphology during the 8-day experiment. On the other hand, neurite extensions are maintained by cells co-treated with PACAP and the neuropoietic cytokine leukemia inhibitory factor (LIF), with similar levels in the number of neurite-bearing cells observed at day 8 to that at day 4. Interestingly, the induction of neurite outgrowths in SH-SY5Y cells requires a minimal 2-h treatment with PACAP, with 4–6 h of treatment inducing similar peak levels of neurite-bearing cells observed at day 4 to that of continual PACAP treatment. LIF is not required until day 4 after which continual treatment with LIF is necessary for neurite maintenance in PACAP-treated cells. The signaling pathways activated by LIF were investigated also. LIF elicits signal transducer and activator of transcription 3 (STAT3) but not STAT1 tyrosine phosphorylation along with a transient activation of the ERK and prolonged activation of phosphatidylinositol-3 kinase (PI3K)/protein kinase B (PKB/Akt) in PACAP-treated and control SH-SY5Y cells. Phosphorylation of serine 727 on STAT3 was detected in PACAP-treated but not control cells, and this was prevented by the MEK-1 inhibitor PD98059. Activation of p38 MAPK or c-jun N-terminal kinase (JNK) were not detected, however, an elevated basal level of phosphorylated p38 MAP kinase was observed in PACAP-treated cells. Neither PD98059 nor siRNA knockdown of STAT3 prevented LIF-mediated maintenance of PACAP-induced neurite extensions. However, the PI3K inhibitor wortmannin was found to effectively block LIF maintenance of neurite outgrowths in PACAP-treated cells. Thus, the ability of LIF to stabilize and maintain PACAP-induced neurite outgrowths requires PI3K but not the STAT3 or ERK signaling pathways in SH-SY5Y cells.

AB - Previously, we have shown that pituitary adenylate cyclase-activating polypeptide (PACAP) induces neuritogenesis in human SH-SY5Y cells through a cAMP-dependent but protein kinase A-independent mechanism involving activation of the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways. However, the morphological changes induced by PACAP are transient, with the increase in number of neurite-bearing cells peaking at day 4 before returning to control cell levels over an 8-day time course. Control cells maintain an undifferentiated neuroblast morphology during the 8-day experiment. On the other hand, neurite extensions are maintained by cells co-treated with PACAP and the neuropoietic cytokine leukemia inhibitory factor (LIF), with similar levels in the number of neurite-bearing cells observed at day 8 to that at day 4. Interestingly, the induction of neurite outgrowths in SH-SY5Y cells requires a minimal 2-h treatment with PACAP, with 4–6 h of treatment inducing similar peak levels of neurite-bearing cells observed at day 4 to that of continual PACAP treatment. LIF is not required until day 4 after which continual treatment with LIF is necessary for neurite maintenance in PACAP-treated cells. The signaling pathways activated by LIF were investigated also. LIF elicits signal transducer and activator of transcription 3 (STAT3) but not STAT1 tyrosine phosphorylation along with a transient activation of the ERK and prolonged activation of phosphatidylinositol-3 kinase (PI3K)/protein kinase B (PKB/Akt) in PACAP-treated and control SH-SY5Y cells. Phosphorylation of serine 727 on STAT3 was detected in PACAP-treated but not control cells, and this was prevented by the MEK-1 inhibitor PD98059. Activation of p38 MAPK or c-jun N-terminal kinase (JNK) were not detected, however, an elevated basal level of phosphorylated p38 MAP kinase was observed in PACAP-treated cells. Neither PD98059 nor siRNA knockdown of STAT3 prevented LIF-mediated maintenance of PACAP-induced neurite extensions. However, the PI3K inhibitor wortmannin was found to effectively block LIF maintenance of neurite outgrowths in PACAP-treated cells. Thus, the ability of LIF to stabilize and maintain PACAP-induced neurite outgrowths requires PI3K but not the STAT3 or ERK signaling pathways in SH-SY5Y cells.

KW - neurite

KW - neuroblastoma cells

KW - signaling pathways

U2 - 10.1007/s12031-009-9324-2

DO - 10.1007/s12031-009-9324-2

M3 - Conference Contribution

VL - 42

SP - 297

EP - 297

JO - Journal of Molecular Neuroscience

T2 - Journal of Molecular Neuroscience

JF - Journal of Molecular Neuroscience

SN - 0895-8696

IS - 3

ER -