• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Sep 1987; 7(9): 3156–3167.
PMCID: PMC367950

Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells.

Abstract

Differential screening of cDNA libraries was used to detect and prepare probes for mRNAs that are regulated in PC12 rat pheochromocytoma cells by long-term (2-week) treatment with nerve growth factor (NGF). In response to NGF, PC12 cells change from a chromaffin cell-like to a sympathetic-neuron-like phenotype. Thus, one aim of this study was to identify NGF-regulated mRNAs that may be associated with the attainment of neuronal properties. Eight NGF-regulated mRNAs are described. Five of these increase 3- to 10-fold and three decrease 2- to 10-fold after long-term NGF exposure. Each mRNA was characterized with respect to the time course of the NGF response, regulation by agents other than NGF, and rat tissue distribution. Partial sequences of the cDNAs were used to search for homologies to known sequences. Homology analysis revealed that one mRNA (increased 10-fold) encodes the peptide thymosin beta 4 and a second mRNA (decreased 2-fold) encodes tyrosine hydroxylase. Another of the increased mRNAs was very abundant in sympathetic ganglia, barely detectable in brain and adrenals, and undetectable in all other tissues surveyed. One of the decreased mRNAs, by contrast, was very abundant in the adrenals and nearly absent in the sympathetic ganglia. With the exception of fibroblast growth factor, which is the only other agent known to mimic the differentiating effects of NGF on PC12 cells, none of the treatments tested (epidermal growth factor, insulin, dibutyryl cyclic AMP, dexamethasone, phorbol ester, and depolarization) reproduced the regulation observed with NGF. These and additional findings suggest that the NGF-regulated mRNAs may play roles in the establishment of the neuronal phenotype and that the probes described here will be useful to study the mechanism of action of NGF and the development and differentiation of neurons.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (3.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson DJ, Axel R. Molecular probes for the development and plasticity of neural crest derivatives. Cell. 1985 Sep;42(2):649–662. [PubMed]
  • Baetge EE, Kaplan BB, Reis DJ, Joh TH. Translation of tyrosine hydroxylase from poly(A)-mRNA in pheochromocytoma cells is enhanced by dexamethasone. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1269–1273. [PMC free article] [PubMed]
  • Biggin MD, Gibson TJ, Hong GF. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. [PMC free article] [PubMed]
  • Bilofsky HS, Burks C, Fickett JW, Goad WB, Lewitter FI, Rindone WP, Swindell CD, Tung CS. The GenBank genetic sequence databank. Nucleic Acids Res. 1986 Jan 10;14(1):1–4. [PMC free article] [PubMed]
  • Bjerre B, Björklund A. The production of catecholamine-containing cells in vitro by young chick embryos: effects of "nerve growth factor" (NGF) and its antiserum. Neurobiology. 1973;3(3):140–161. [PubMed]
  • Black IB, Chikaraishi DM, Lewis EJ. Trans-synaptic increase in RNA coding for tyrosine hydroxylase in a rat sympathetic ganglion. Brain Res. 1985 Jul 22;339(1):151–153. [PubMed]
  • Burstein DE, Greene LA. Evidence for RNA synthesis-dependent and -independent pathways in stimulation of neurite outgrowth by nerve growth factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6059–6063. [PMC free article] [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Cochran BH, Reffel AC, Stiles CD. Molecular cloning of gene sequences regulated by platelet-derived growth factor. Cell. 1983 Jul;33(3):939–947. [PubMed]
  • Connolly JL, Green SA, Greene LA. Comparison of rapid changes in surface morphology and coated pit formation of PC12 cells in response to nerve growth factor, epidermal growth factor, and dibutyryl cyclic AMP. J Cell Biol. 1984 Feb;98(2):457–465. [PMC free article] [PubMed]
  • Cremins J, Wagner JA, Halegoua S. Nerve growth factor action is mediated by cyclic AMP- and Ca+2/phospholipid-dependent protein kinases. J Cell Biol. 1986 Sep;103(3):887–893. [PMC free article] [PubMed]
  • Curran T, Morgan JI. Superinduction of c-fos by nerve growth factor in the presence of peripherally active benzodiazepines. Science. 1985 Sep 20;229(4719):1265–1268. [PubMed]
  • Curran T, Peters G, Van Beveren C, Teich NM, Verma IM. FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA. J Virol. 1982 Nov;44(2):674–682. [PMC free article] [PubMed]
  • Dichter MA, Tischler AS, Greene LA. Nerve growth factor-induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature. 1977 Aug 11;268(5620):501–504. [PubMed]
  • Dickson G, Prentice H, Julien JP, Ferrari G, Leon A, Walsh FS. Nerve growth factor activates Thy-1 and neurofilament gene transcription in rat PC12 cells. EMBO J. 1986 Dec 20;5(13):3449–3453. [PMC free article] [PubMed]
  • Doupe AJ, Landis SC, Patterson PH. Environmental influences in the development of neural crest derivatives: glucocorticoids, growth factors, and chromaffin cell plasticity. J Neurosci. 1985 Aug;5(8):2119–2142. [PubMed]
  • Doupe AJ, Patterson PH, Landis SC. Small intensely fluorescent cells in culture: role of glucocorticoids and growth factors in their development and interconversions with other neural crest derivatives. J Neurosci. 1985 Aug;5(8):2143–2160. [PubMed]
  • Dworkin MB, Dawid IB. Use of a cloned library for the study of abundant poly(A)+RNA during Xenopus laevis development. Dev Biol. 1980 May;76(2):449–464. [PubMed]
  • Edgar DH, Thoenen H. Selective enzyme induction in a nerve growth factor-responsive pheochromocytoma cell line (PC 12). Brain Res. 1978 Oct 6;154(1):186–190. [PubMed]
  • Feinberg AP, Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. [PubMed]
  • Feinstein SC, Dana SL, McConlogue L, Shooter EM, Coffino P. Nerve growth factor rapidly induces ornithine decarboxylase mRNA in PC12 rat pheochromocytoma cells. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5761–5765. [PMC free article] [PubMed]
  • Friedman RL, Manly SP, McMahon M, Kerr IM, Stark GR. Transcriptional and posttranscriptional regulation of interferon-induced gene expression in human cells. Cell. 1984 Oct;38(3):745–755. [PubMed]
  • Goedert M, Otten U, Hunt SP, Bond A, Chapman D, Schlumpf M, Lichtensteiger W. Biochemical and anatomical effects of antibodies against nerve growth factor on developing rat sensory ganglia. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1580–1584. [PMC free article] [PubMed]
  • Goodall GJ, Richardson M, Furuichi Y, Wodnar-Filipowicz A, Horecker BL. Sequence of a cloned 523-bp cDNA for thymosin beta 4. Arch Biochem Biophys. 1985 Jan;236(1):445–447. [PubMed]
  • Greenberg ME, Greene LA, Ziff EB. Nerve growth factor and epidermal growth factor induce rapid transient changes in proto-oncogene transcription in PC12 cells. J Biol Chem. 1985 Nov 15;260(26):14101–14110. [PubMed]
  • Greenberg ME, Ziff EB. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. [PubMed]
  • Greenberg ME, Ziff EB, Greene LA. Stimulation of neuronal acetylcholine receptors induces rapid gene transcription. Science. 1986 Oct 3;234(4772):80–83. [PubMed]
  • Greene LA, Tischler AS. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. [PMC free article] [PubMed]
  • Grima B, Lamouroux A, Blanot F, Biguet NF, Mallet J. Complete coding sequence of rat tyrosine hydroxylase mRNA. Proc Natl Acad Sci U S A. 1985 Jan;82(2):617–621. [PMC free article] [PubMed]
  • Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L. Isolation and characterization of full-length cDNA clones for human alpha-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol. 1983 May;3(5):787–795. [PMC free article] [PubMed]
  • Halegoua S, Patrick J. Nerve growth factor mediates phosphorylation of specific proteins. Cell. 1980 Nov;22(2 Pt 2):571–581. [PubMed]
  • Hannappel E, Leibold W. Biosynthesis rates and content of thymosin beta 4 in cell lines. Arch Biochem Biophys. 1985 Jul;240(1):236–241. [PubMed]
  • Hannappel E, Xu GJ, Morgan J, Hempstead J, Horecker BL. Thymosin beta 4: a ubiquitous peptide in rat and mouse tissues. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2172–2175. [PMC free article] [PubMed]
  • Hedrick SM, Cohen DI, Nielsen EA, Davis MM. Isolation of cDNA clones encoding T cell-specific membrane-associated proteins. Nature. 1984 Mar 8;308(5955):149–153. [PubMed]
  • Huff KR, Guroff G. Nerve growth factor-induced reduction in epidermal growth factor responsiveness and epidermal growth factor receptors in PC12 cells: an aspect of cell differentiation. Biochem Biophys Res Commun. 1979 Jul 12;89(1):175–180. [PubMed]
  • Lamouroux A, Faucon Biguet N, Samolyk D, Privat A, Salomon JC, Pujol JF, Mallet J. Identification of cDNA clones coding for rat tyrosine hydroxylase antigen. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3881–3885. [PMC free article] [PubMed]
  • Levi A, Eldridge JD, Paterson BM. Molecular cloning of a gene sequence regulated by nerve growth factor. Science. 1985 Jul 26;229(4711):393–395. [PubMed]
  • Levi-Montalcini R, Angeletti PU. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. [PubMed]
  • Lewis EJ, Tank AW, Weiner N, Chikaraishi DM. Regulation of tyrosine hydroxylase mRNA by glucocorticoid and cyclic AMP in a rat pheochromocytoma cell line. Isolation of a cDNA clone for tyrosine hydroxylase mRNA. J Biol Chem. 1983 Dec 10;258(23):14632–14637. [PubMed]
  • Linzer DI, Lee SJ, Ogren L, Talamantes F, Nathans D. Identification of proliferin mRNA and protein in mouse placenta. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4356–4359. [PMC free article] [PubMed]
  • Linzer DI, Nathans D. Growth-related changes in specific mRNAs of cultured mouse cells. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4271–4275. [PMC free article] [PubMed]
  • Linzer DI, Nathans D. Nucleotide sequence of a growth-related mRNA encoding a member of the prolactin-growth hormone family. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4255–4259. [PMC free article] [PubMed]
  • Low TL, Hu SK, Goldstein AL. Complete amino acid sequence of bovine thymosin beta 4: a thymic hormone that induces terminal deoxynucleotidyl transferase activity in thymocyte populations. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1162–1166. [PMC free article] [PubMed]
  • McGuire JC, Greene LA. Rapid stimulation by nerve growth factor of amino acid uptake by clonal PC12 pheochromocytoma cells. J Biol Chem. 1979 May 10;254(9):3362–3367. [PubMed]
  • McMahon M, Stark GR, Kerr IM. Interferon-induced gene expression in wild-type and interferon-resistant human lymphoblastoid (Daudi) cells. J Virol. 1986 Jan;57(1):362–366. [PMC free article] [PubMed]
  • Messing J, Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. [PubMed]
  • Mobley WC, Rutkowski JL, Tennekoon GI, Buchanan K, Johnston MV. Choline acetyltransferase activity in striatum of neonatal rats increased by nerve growth factor. Science. 1985 Jul 19;229(4710):284–287. [PubMed]
  • Mobley WC, Schenker A, Shooter EM. Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry. 1976 Dec 14;15(25):5543–5552. [PubMed]
  • Morrison RS, Sharma A, de Vellis J, Bradshaw RA. Basic fibroblast growth factor supports the survival of cerebral cortical neurons in primary culture. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7537–7541. [PMC free article] [PubMed]
  • Moscatelli D, Presta M, Rifkin DB. Purification of a factor from human placenta that stimulates capillary endothelial cell protease production, DNA synthesis, and migration. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2091–2095. [PMC free article] [PubMed]
  • Okayama H, Berg P. High-efficiency cloning of full-length cDNA. Mol Cell Biol. 1982 Feb;2(2):161–170. [PMC free article] [PubMed]
  • Otten U, Towbin M. Permissive action of glucocorticoids in induction of tyrosine hydroxylase by nerve growth factor in a pheochromocytoma cell line. Brain Res. 1980 Jul 7;193(1):304–308. [PubMed]
  • Richter-Landsberg C, Greene LA, Shelanski ML. Cell surface Thy-1-cross-reactive glycoprotein in cultured PC12 cells: modulation by nerve growth factor and association with the cytoskeleton. J Neurosci. 1985 Feb;5(2):468–476. [PubMed]
  • Rowekamp W, Firtel RA. Isolation of developmentally regulated genes from Dictyostelium. Dev Biol. 1980 Oct;79(2):409–418. [PubMed]
  • Sanger F, Coulson AR. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. [PubMed]
  • Sanger F, Coulson AR, Barrell BG, Smith AJ, Roe BA. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Schubert D, LaCorbiere M, Klier FG, Steinbach JH. The modulation of neurotransmitter synthesis by steroid hormones and insulin. Brain Res. 1980 May 19;190(1):67–79. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Togari A, Baker D, Dickens G, Guroff G. The neurite-promoting effect of fibroblast growth factor on PC12 cells. Biochem Biophys Res Commun. 1983 Aug 12;114(3):1189–1193. [PubMed]
  • Togari A, Dickens G, Kuzuya H, Guroff G. The effect of fibroblast growth factor on PC12 cells. J Neurosci. 1985 Feb;5(2):307–316. [PubMed]
  • Unsicker K, Krisch B, Otten U, Thoenen H. Nerve growth factor-induced fiber outgrowth from isolated rat adrenal chromaffin cells: impairment by glucocorticoids. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3498–3502. [PMC free article] [PubMed]
  • Walicke P, Cowan WM, Ueno N, Baird A, Guillemin R. Fibroblast growth factor promotes survival of dissociated hippocampal neurons and enhances neurite extension. Proc Natl Acad Sci U S A. 1986 May;83(9):3012–3016. [PMC free article] [PubMed]
  • Williams JG, Lloyd MM. Changes in the abundance of polyadenylated RNA during slime mould development measured using cloned molecular hybridization probes. J Mol Biol. 1979 Mar 25;129(1):19–35. [PubMed]
  • Wodnar-Filipowicz A, Gubler U, Furuichi Y, Richardson M, Nowoswiat EF, Poonian MS, Horecker BL. Cloning and sequence analysis of cDNA for rat spleen thymosin beta 4. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2295–2297. [PMC free article] [PubMed]
  • Xu GJ, Hannappel E, Morgan J, Hempstead J, Horecker BL. Synthesis of thymosin beta 4 by peritoneal macrophages and adherent spleen cells. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4006–4009. [PMC free article] [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • Gene
    Gene
    Gene links
  • Gene (nucleotide)
    Gene (nucleotide)
    Records in Gene identified from shared sequence links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...