scholarly journals Mutants of PC12 cells with altered cyclic AMP responses.

1984 ◽  
Vol 4 (10) ◽  
pp. 2091-2097 ◽  
Author(s):  
T Block ◽  
C Kon ◽  
B M Breckenridge
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Pc12 Cells ◽  
Substrate Concentration ◽  
Morphological Traits ◽  
Phosphodiesterase Activity ◽  
Camp Metabolism ◽  
Coupling Protein

PC12 cells, derived from a rat pheochromocytoma, were mutagenized and selected in media containing agents known to elevate intracellular concentrations of cyclic AMP (cAMP). More than 40 clones were isolated by selection with cholera toxin or 2-chloroadenosine or both. The variants that were deficient in accumulating cAMP were obtained by using a protocol in which 1 microM 8-bromo-cAMP was included in addition to the agonist. Certain of these variants were partially characterized with respect to the site of altered cAMP metabolism. The profiles of adenylate cyclase activity responsiveness of certain variants to guanosine-5'-(beta, gamma-imido) triphosphate and to forskolin resembled those of UNC and cyc phenotypes of S49 lymphoma cells, which are functionally deficient in the GTP-sensitive coupling protein, Ns. Other variants were characterized by increased cyclic nucleotide phosphodiesterase activity at low substrate concentration. Diverse morphological traits were observed among the variants, but it was not possible to assign them to a particular cAMP phenotype. Two revertants of a PC12 mutant were isolated and observed to have regained a cellular cAMP response to 2-chloroadenosine and to forskolin. It is hoped that these PC12 mutants will have utility for defining cAMP-mediated functions, including any links to the action of nerve growth factor, in cells derived from the neural crest.

Mutants of PC12 cells with altered cyclic AMP responses

1984 ◽  
Vol 4 (10) ◽  
pp. 2091-2097
Author(s):  
T Block ◽  
C Kon ◽  
B M Breckenridge
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Pc12 Cells ◽  
Substrate Concentration ◽  
Morphological Traits ◽  
Phosphodiesterase Activity ◽  
Camp Metabolism ◽  
Coupling Protein

PC12 cells, derived from a rat pheochromocytoma, were mutagenized and selected in media containing agents known to elevate intracellular concentrations of cyclic AMP (cAMP). More than 40 clones were isolated by selection with cholera toxin or 2-chloroadenosine or both. The variants that were deficient in accumulating cAMP were obtained by using a protocol in which 1 microM 8-bromo-cAMP was included in addition to the agonist. Certain of these variants were partially characterized with respect to the site of altered cAMP metabolism. The profiles of adenylate cyclase activity responsiveness of certain variants to guanosine-5'-(beta, gamma-imido) triphosphate and to forskolin resembled those of UNC and cyc phenotypes of S49 lymphoma cells, which are functionally deficient in the GTP-sensitive coupling protein, Ns. Other variants were characterized by increased cyclic nucleotide phosphodiesterase activity at low substrate concentration. Diverse morphological traits were observed among the variants, but it was not possible to assign them to a particular cAMP phenotype. Two revertants of a PC12 mutant were isolated and observed to have regained a cellular cAMP response to 2-chloroadenosine and to forskolin. It is hoped that these PC12 mutants will have utility for defining cAMP-mediated functions, including any links to the action of nerve growth factor, in cells derived from the neural crest.

scholarly journals Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) and Nerve Growth Factor (NGF) Induce PACAP Expression in PC12 Cells

1998 ◽  
Vol 76 ◽  
pp. 171
Author(s):  
Nami Hagihara ◽  
Hitoshi Hashimoto ◽  
Kyohei Yamamoto ◽  
Takashi Fujita ◽  
Norihito Shintani ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Adenylate Cyclase ◽  
Pc12 Cells ◽  
Nerve Growth ◽  
Pituitary Adenylate Cyclase

Mechanisms of apoptosis in PC12 cells irreversibly differentiated with nerve growth factor and cyclic AMP

1999 ◽  
Vol 821 (1) ◽  
pp. 60-68 ◽  
Author(s):  
Nathalie Lambeng ◽  
Patrick P Michel ◽  
Bernard Brugg ◽  
Yves Agid ◽  
Merle Ruberg
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Pc12 Cells ◽  
Nerve Growth

scholarly journals Structure of the gene encoding VGF, a nervous system-specific mRNA that is rapidly and selectively induced by nerve growth factor in PC12 cells.

1991 ◽  
Vol 11 (5) ◽  
pp. 2335-2349 ◽  
Author(s):  
S R Salton ◽  
D J Fischberg ◽  
K W Dong
Keyword(s):  
Nervous System ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cyclic Amp ◽  
Pc12 Cells ◽  
Cdna Clone ◽  
Neural Differentiation ◽  
Epidermal Growth ◽  
Specific Mrna

Nerve growth factor (NGF) plays a critical role in the development and survival of neurons in the peripheral nervous system. Following treatment with NGF but not epidermal growth factor, rat pheochromocytoma (PC12) cells undergo neural differentiation. We have cloned a nervous system-specific mRNA, NGF33.1, that is rapidly and relatively selectively induced by treatment of PC12 cells with NGF and basic fibroblast growth factor in comparison with epidermal growth factor. Analysis of the nucleic acid and predicted amino acid sequences of the NGF33.1 cDNA clone suggested that this clone corresponded to the NGF-inducible mRNA called VGF (A. Levi, J. D. Eldridge, and B. M. Paterson, Science 229:393-395, 1985; R. Possenti, J. D. Eldridge, B. M. Paterson, A. Grasso, and A. Levi, EMBO J. 8:2217-2223, 1989). We have used the NGF33.1 cDNA clone to isolate and characterize the VGF gene, and in this paper we report the complete sequence of the VGF gene, including 853 bases of 5' flank revealed TATAA and CCAAT elements, several GC boxes, and a consensus cyclic AMP response element-binding protein binding site. The VGF promoter contains sequences homologous to other NGF-inducible, neuronal promoters. We further show that VGF mRNA is induced in PC12 cells to a greater extent by depolarization and by phorbol-12-myristate-13-acetate treatment than by 8-bromo-cyclic AMP treatment. By Northern (RNA) and RNase protection analysis, VGF mRNA is detectable in embryonic and postnatal central and peripheral nervous tissues but not in a number of nonneural tissues. In the cascade of events which ultimately leads to the neural differentiation of NGF-treated PC12 cells, the VGF gene encodes the most rapidly and selectively regulated, nervous-system specific mRNA yet identified.

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