NF-κB Signaling Promotes Both Cell Survival and Neurite Process Formation in Nerve Growth Factor-Stimulated PC12 Cells

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.

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Synergistic effects of nerve growth factor and phorbol 12-myristate 13-acetate on rapid motility and process formation in PC12 cells: The role of laminin

Journal of Neuroscience Research ◽

10.1002/jnr.490310207 ◽

1992 ◽

Vol 31 (2) ◽

pp. 263-272 ◽

Cited By ~ 9

Author(s):

D. Glowacka ◽

D. D. Ginty ◽

J. A. Wagner

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Pc12 Cells ◽

Synergistic Effects ◽

Nerve Growth ◽

Process Formation

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Analysis of the Role of Nerve Growth Factor in Promoting Cell Survival During Endoplasmic Reticulum Stress in PC12 Cells

The Unfolded Protein Response and Cellular Stress, Part B - Methods in Enzymology ◽

10.1016/b978-0-12-385114-7.00003-9 ◽

2011 ◽

pp. 53-70 ◽

Cited By ~ 10

Author(s):

Koji Shimoke ◽

Harue Sasaya ◽

Toshihiko Ikeuchi

Keyword(s):

Endoplasmic Reticulum ◽

Nerve Growth Factor ◽

Growth Factor ◽

Endoplasmic Reticulum Stress ◽

Cell Survival ◽

Pc12 Cells ◽

Nerve Growth

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Dehydroepiandrosterone Induces a Neuroendocrine Phenotype in Nerve Growth Factor-Stimulated Chromaffin Pheochromocytoma PC12 Cells

Endocrinology ◽

10.1210/en.2007-0645 ◽

2007 ◽

Vol 149 (1) ◽

pp. 320-328 ◽

Cited By ~ 20

Author(s):

Christian G. Ziegler ◽

Flavie Sicard ◽

Peter Lattke ◽

Stefan R. Bornstein ◽

Monika Ehrhart-Bornstein ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Cell Survival ◽

Pc12 Cells ◽

Adrenal Cortex ◽

Chromaffin Cell ◽

Hydroxylase Deficiency ◽

Neuronal Marker ◽

Nerve Growth ◽

21 Hydroxylase Deficiency

The adrenal androgen dehydroepiandrosterone (DHEA) is produced in the inner zone of the adrenal cortex, which is in direct contact to adrenal medullary cells. Due to their close anatomical proximity and tightly intermingled cell borders, a direct interaction of adrenal cortex and medulla has been postulated. In humans congenital adrenal hyperplasia due to 21-hydroxylase deficiency results in androgen excess accompanied by severe adrenomedullary dysplasia and chromaffin cell dysfunction. Therefore, to define the mechanisms of DHEA action on chromaffin cell function, we investigated its effect on cell survival and differentiation processes on a molecular level in the chromaffin cell line PC12. DHEA lessened the positive effect of NGF on cell survival and neuronal differentiation. Nerve growth factor (NGF)-mediated induction of a neuronal phenotype was inhibited by DHEA as indicated by reduced neurite outgrowth and decreased expression of neuronal marker proteins such as synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein-2. We examined whether DHEA may stimulate the cells toward a neuroendocrine phenotype. DHEA significantly elevated catecholamine release from unstimulated PC12 cells in the presence but not absence of NGF. Accordingly, DHEA enhanced the expression of the neuroendocrine marker protein chromogranin A. Next, we explored the possible molecular mechanisms of DHEA and NGF interaction. We demonstrate that NGF-induced ERK1/2 phosphorylation was reduced by DHEA. In summary, our data show that DHEA influences cell survival and differentiation processes in PC12 cells, possibly by interacting with the ERK1/2 MAPK pathway. DHEA drives NGF-stimulated cells toward a neuroendocrine phenotype, suggesting that the interaction of intraadrenal steroids and growth factors is required for the maintenance of an intact adrenal medulla.

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Neurite Extension Occurs in the Absence of Regulated Exocytosis in PC12 Subclones

Molecular Biology of the Cell ◽

10.1091/mbc.10.9.2919 ◽

1999 ◽

Vol 10 (9) ◽

pp. 2919-2931 ◽

Cited By ~ 33

Author(s):

Chiara Leoni ◽

Andrea Menegon ◽

Fabio Benfenati ◽

Daniela Toniolo ◽

Maria Pennuto ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Pc12 Cells ◽

Growth Factor Receptor ◽

Morphological Differentiation ◽

Nerve Growth ◽

Neurite Extension ◽

Process Formation ◽

Dense Core Granules ◽

Activation Pathways

We have investigated the process leading to differentiation of PC12 cells. This process is known to include extension of neurites and changes in the expression of subsets of proteins involved in cytoskeletal rearrangements or in neurosecretion. To this aim, we have studied a PC12 clone (trk-PC12) stably transfected with the nerve growth factor receptor TrkA. These cells are able to undergo both spontaneous and neurotrophin-induced morphological differentiation. However, both undifferentiated and nerve growth factor-differentiated trk-PC12 cells appear to be completely defective in the expression of proteins of the secretory apparatus, including proteins of synaptic vesicles and large dense-core granules, neurotransmitter transporters, and neurotransmitter-synthesizing enzymes. These results indicate that neurite extension can occur independently of the presence of the neurosecretory machinery, including the proteins that constitute the fusion machine, suggesting the existence of differential activation pathways for the two processes during neuronal differentiation. These findings have been confirmed in independent clones obtained from PC12-27, a previously characterized PC12 variant clone globally incompetent for regulated secretion. In contrast, the integrity of the Rab cycle appears to be necessary for neurite extension, because antisense oligonucleotides against the neurospecific isoform of Rab-guanosine diphosphate-dissociation inhibitor significantly interfere with process formation.

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