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

1999 ◽  
Vol 10 (9) ◽  
pp. 2919-2931 ◽  
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.

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

2000 ◽  
Vol 20 (20) ◽  
pp. 7556-7563 ◽  
Author(s):  
Erik D. Foehr ◽  
Xin Lin ◽  
Alison O'Mahony ◽  
Romas Geleziunas ◽  
Ralph A. Bradshaw ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Survival ◽  
Pc12 Cells ◽  
Nerve Growth ◽  
Process Formation

scholarly journals Lateral diffusion of nerve growth factor receptor: modulation by ligand-binding and cell-associated factors.

1990 ◽  
Vol 1 (8) ◽  
pp. 605-614 ◽  
Author(s):  
G Venkatakrishnan ◽  
C A McKinnon ◽  
A H Ross ◽  
D E Wolf
Keyword(s):  
Monoclonal Antibody ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Line ◽  
Pc12 Cells ◽  
Growth Factor Receptor ◽  
Lateral Diffusion ◽  
Nerve Growth Factor Receptor ◽  
Nerve Growth ◽  
Cos Cells

We compared the properties in human melanoma cell line A875 and rat pheochromocytoma cell line PC12 of nerve growth factor receptor (NGFr). We also analyzed NGFr and a truncated NGFR lacking the cytoplasmic domain, which were transiently expressed in COS cells. The full-length NGFR expressed in COS cells bound nerve growth factor (NGF) with positive cooperativity, but A875 NGFr and truncated NGFr in COS cells did not display positive cooperativity. The anti-human NGFr monoclonal antibody NGFR5 was characterized and found not to compete with NGF for binding to NGFr. Fabs were prepared from NGFR5 and 192, an anti-rat NGFR monoclonal antibody that was previously shown not to compete with NGF for binding. Fluorescein-labeled Fabs were used to measure the distribution and lateral diffusion of the NGFr. NGFr expressed on COS and A875 cells are diffusely distributed, but NGFr on the surface of PC12 cells appeared, for some cells, to be patched. In A875 cells, 51% of the NGFr was free to diffuse with diffusion coefficient (D) approximately 7 X 10(-10) cm2/s. In COS cells, 43% diffused with D approximately 5 X 10(-10) cm2/s. There was no significant difference in diffusibility between the full-length NGFr and the truncated NGFr. We compared NGFr diffusion on PC12 cells in suspension or adherent to collagen-coated coverslips. For suspension cells, we obtained 32% recovery with D approximately 2.5 X 10(-9) cm2/s. On adherent cells, we obtained 17% recovery with 6 X 10(-9) cm2/s. Binding of NGF enhanced lateral diffusion of NGFr in A875 cells and in PC12 cells in suspension but did not alter lateral diffusion of NGFr in COS cells or in adherent PC12 cells. NGF had no effect on the diffusing fraction or the distribution of NGFR for any cell line.

Decreased levels of nerve growth factor receptor on dexamethasone-treated PC12 cells

1988 ◽  
Vol 20 (4) ◽  
pp. 411-419 ◽  
Author(s):  
Maria D. Tocco ◽  
M. L. Contreras ◽  
S. Koizumi ◽  
G. Dickens ◽  
G. Guroff
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Pc12 Cells ◽  
Growth Factor Receptor ◽  
Nerve Growth Factor Receptor ◽  
Nerve Growth
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