Involvement of cAMP in nerve growth factor-triggered p35/Cdk5 activation and differentiation in PC12 cells

2010 ◽  
Vol 299 (2) ◽  
pp. C516-C527 ◽  
Author(s):  
Mei-Chih Chen ◽  
Ho Lin ◽  
Fu-Ning Hsu ◽  
Pao-Hsuan Huang ◽  
Guan-Shun Lee ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Adenylyl Cyclase ◽  
Pc12 Cells ◽  
Intracellular Camp ◽  
Camp Production ◽  
Nerve Growth ◽  
Neurite Initiation ◽  
Pc12 Differentiation

The signaling mechanisms underlying cell differentiation have been extensively studied with the use of rat PC12 cells as a model system. Nerve growth factor (NGF) is a trophic factor inducing PC12 cell differentiation through the activation of the p35/cyclin-dependent kinase 5 (Cdk5) complex. It has been reported that adenylyl cyclase activation and cAMP production may be involved in NGF-dependent actions. Our previous results indicate that cAMP activates the p35/Cdk5 complex in reproductive cells. Therefore, the role of cAMP in NGF-triggered p35/Cdk5 activation and PC12 differentiation was interesting to explore. Our results indicate that roscovitine, a molecular inhibitor of Cdk5, blocks cAMP-triggered PC12 differentiation, which was evaluated by neurite initiation, a decrease in proliferation, and cell cycle G1 arrest. The following data show that cAMP treatment increased Cdk5 activity through p35 upregulation. cAMP downstream components, protein kinase A (PKA) and phosphorylated cAMP response element binding protein (CREB), are involved in this regulation. The immunocytochemical results indicate that PKA inhibition disrupted cAMP-triggered p35/Cdk5 localization in PC12 cells. In addition, adenylyl cyclase inhibition was found to diminish NGF-induced intracellular cAMP production, CREB phosphorylation, and p35 expression. The cAMP antagonist and the PKA inhibitors reduced NGF-induced p35 expression. Finally, NGF-triggered PC12 differentiation was partially decreased by adenylyl cyclase or PKA inhibitors. In conclusion, these results demonstrate that cAMP may play a role in NGF-p35/Cdk5-dependent PC12 differentiation.

scholarly journals The role of cAMP in nerve growth factor-promoted neurite outgrowth in PC12 cells.

1986 ◽  
Vol 102 (3) ◽  
pp. 821-829 ◽  
Author(s):  
C Richter-Landsberg ◽  
B Jastorff
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neurite Outgrowth ◽  
Pc12 Cells ◽  
Morphological Differentiation ◽  
Intracellular Camp ◽  
Nerve Growth ◽  
Process Formation ◽  
Dependent Protein

Nerve growth factor (NGF)-mediated neurite outgrowth in rat pheochromocytoma PC12 cells has been described to be synergistically potentiated by the simultaneous addition of dibutyryl cAMP. To elucidate further the role of cAMP in NGF-induced neurite outgrowth we have used the adenylate cyclase activator forskolin, cAMP, and a set of chemically modified cAMP analogues, including the adenosine cyclic 3',5'-phosphorothioates (cAMPS) (Rp)-cAMPS and (Sp)-cAMPS. These diastereomers have differential effects on the activation of cAMP-dependent protein kinases, i.e., (Sp)-cAMPS behaves as a cAMP agonist and (Rp)-cAMPS behaves as a cAMP antagonist. Our data show that the establishment of a neuritic network, as observed from PC12 cells treated with NGF alone, could not be induced by either forskolin, cAMP, or cAMP analogues alone. The presence of NGF in combination with forskolin or cAMP or its agonistic analogues potentiated the initiation of neurite outgrowth from PC12 cells. The (Sp)-cAMPS-induced stimulation of NGF-mediated process formation was successfully blocked by the (Rp)-cAMPS diastereomer. On the other hand, NGF-stimulated neurite outgrowth was not inhibited by the presence of the cAMP antagonist (Rp)-cAMPS. We conclude that the morphological differentiation of PC12 cells stimulated by NGF does not require cAMP as a second messenger. The constant increase of intracellular cAMP, caused by either forskolin or cAMP and the analogues, in combination with NGF, not only rapidly stimulated early neurite outgrowth but also exerted a maintaining effect on the neuronal network established by NGF.

scholarly journals Nerve Growth Factor-Induced Differentiation of PC12 Cells Is Accompanied by Elevated Adenylyl Cyclase Activity

Neurosignals ◽  
2010 ◽  
Vol 18 (1) ◽  
pp. 32-42 ◽  
Author(s):  
H.S. Yung ◽  
K.H. Lai ◽  
K.B.S. Chow ◽  
N.Y. Ip ◽  
K.W.K. Tsim ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Adenylyl Cyclase ◽  
Pc12 Cells ◽  
Cyclase Activity ◽  
Adenylyl Cyclase Activity ◽  
Induced Differentiation ◽  
Nerve Growth

scholarly journals E2F4 Actively Promotes the Initiation and Maintenance of Nerve Growth Factor-Induced Cell Differentiation

1999 ◽  
Vol 19 (9) ◽  
pp. 6048-6056 ◽  
Author(s):  
Stephan P. Persengiev ◽  
Ivanela I. Kondova ◽  
Daniel L. Kilpatrick
Keyword(s):  
Cell Differentiation ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neuronal Differentiation ◽  
Pc12 Cells ◽  
Pc12 Cell ◽  
Growth Regulation ◽  
Critical Role ◽  
Terminal Differentiation ◽  
Nerve Growth

ABSTRACT E2F transcription factors play a critical role in cell cycle progression through the regulation of genes required for G1/S transition. They are also thought to be important for growth arrest; however, their potential role in the cell differentiation process has not been previously examined. Here, we demonstrate that E2F4 is highly upregulated following the neuronal differentiation of PC12 cells with nerve growth factor (NGF), while E2F1, E2F3, and E2F5 are downregulated. Immunoprecipitation and subcellular fractionation studies demonstrated that both the nuclear localization of E2F4 and its association with the Rb family member p130 increased following neuronal differentiation. The forced expression of E2F4 markedly enhanced the rate of PC12 cell differentiation induced by NGF and also greatly lowered the rate at which cells lost their neuronal phenotype following NGF removal. Importantly, this effect occurred in the absence of any significant change in the growth regulation of PC12 cells by NGF. Further, the downregulation of E2F4 expression with antisense oligodeoxynucleotides inhibited NGF-induced neurite outgrowth, indicating an important role for this factor during PC12 cell differentiation. Finally, E2F4 expression was found to increase dramatically in the developing rat cerebral cortex and cerebellum, as neuroblasts became postmitotic and initiated terminal differentiation. These findings demonstrate that, in addition to its effects on cell proliferation, E2F4 actively promotes the neuronal differentiation of PC12 cells as well as the retention of this state. Further, this effect is independent of alterations in cell growth and may involve interactions between E2F4 and the neuronal differentiation program itself. E2F4 may be an important participant in the terminal differentiation of neuroblasts.

scholarly journals Nerve growth factor stimulates phosphorylation of phospholipase C-gamma in PC12 cells

1991 ◽  
Vol 266 (3) ◽  
pp. 1359-1362 ◽  
Author(s):  
U H Kim ◽  
D Fink ◽  
H S Kim ◽  
D J Park ◽  
M L Contreras ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Phospholipase C ◽  
Pc12 Cells ◽  
Nerve Growth ◽  
Phospholipase C Gamma
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