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PMID: 3670309 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 7 ·No. 9 ·1987-09-00 ·Pages 3156-67

Leonard DG, Ziff EB, Greene LA

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.

MeSH Terms
Animals Cell Differentiation Cell Line Chromaffin System/cytology,physiology DNA/genetics Gene Expression Regulation Nerve Growth Factors/physiology Pheochromocytoma RNA, Messenger/genetics Rats Sympathetic Nervous System/cytology,physiology Time Factors Tissue Distribution
Chemicals
Nerve Growth Factors RNA, Messenger DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Leonard D G
Department of Biochemistry, New York University School of Medicine, New York 10016.
Ziff E B
Greene L A
References (66)
66 references, click to expand
  1. Nerve growth factor activates Thy-1 and neurofilament gene transcription in rat PC12 cells.
    EMBO J. 1986 Dec 20;5(13):3449-53 PMID: 16453742
  2. 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):2143-60 PMID: 4020433
  3. High-efficiency cloning of full-length cDNA.
    Mol Cell Biol. 1982 Feb;2(2):161-70 PMID: 6287227
  4. Molecular cloning of gene sequences regulated by platelet-derived growth factor.
    Cell. 1983 Jul;33(3):939-47 PMID: 6872001
  5. Fibroblast growth factor promotes survival of dissociated hippocampal neurons and enhances neurite extension.
    Proc Natl Acad Sci U S A. 1986 May;83(9):3012-6 PMID: 3458259
  6. Isolation of cDNA clones encoding T cell-specific membrane-associated proteins.
    Nature. 1984 Mar 8-14;308(5955):149-53 PMID: 6199676
  7. The neurite-promoting effect of fibroblast growth factor on PC12 cells.
    Biochem Biophys Res Commun. 1983 Aug 12;114(3):1189-93 PMID: 6225431
  8. Identification of cDNA clones coding for rat tyrosine hydroxylase antigen.
    Proc Natl Acad Sci U S A. 1982 Jun;79(12):3881-5 PMID: 6179090
  9. The modulation of neurotransmitter synthesis by steroid hormones and insulin.
    Brain Res. 1980 May 19;190(1):67-79 PMID: 6103732
  10. Complete coding sequence of rat tyrosine hydroxylase mRNA.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):617-21 PMID: 2857492
  11. 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 PMID: 376855
  12. Interferon-induced gene expression in wild-type and interferon-resistant human lymphoblastoid (Daudi) cells.
    J Virol. 1986 Jan;57(1):362-6 PMID: 2934555
  13. 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):175-80 PMID: 314289
  14. 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):140-61 PMID: 4723316
  15. Rapid stimulation by nerve growth factor of amino acid uptake by clonal PC12 pheochromocytoma cells.
    J Biol Chem. 1979 May 10;254(9):3362-7 PMID: 429355
  16. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum.
    Anal Biochem. 1984 Feb;137(1):266-7 PMID: 6329026
  17. Cloning and sequence analysis of cDNA for rat spleen thymosin beta 4.
    Proc Natl Acad Sci U S A. 1984 Apr;81(8):2295-7 PMID: 6201851
  18. Transcriptional and posttranscriptional regulation of interferon-induced gene expression in human cells.
    Cell. 1984 Oct;38(3):745-55 PMID: 6548414
  19. Nerve growth factor rapidly induces ornithine decarboxylase mRNA in PC12 rat pheochromocytoma cells.
    Proc Natl Acad Sci U S A. 1985 Sep;82(17):5761-5 PMID: 3862093
  20. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.
    Gene. 1982 Oct;19(3):269-76 PMID: 6295880
  21. Nerve growth factor-induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line.
    Nature. 1977 Aug 11;268(5620):501-4 PMID: 329148
  22. 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):304-8 PMID: 6103743
  23. Synthesis of thymosin beta 4 by peritoneal macrophages and adherent spleen cells.
    Proc Natl Acad Sci U S A. 1982 Jul;79(13):4006-9 PMID: 6955787
  24. Molecular probes for the development and plasticity of neural crest derivatives.
    Cell. 1985 Sep;42(2):649-62 PMID: 3839717
  25. FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA.
    J Virol. 1982 Nov;44(2):674-82 PMID: 6292525
  26. Identification of proliferin mRNA and protein in mouse placenta.
    Proc Natl Acad Sci U S A. 1985 Jul;82(13):4356-9 PMID: 3859868
  27. 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):14632-7 PMID: 6139379
  28. Biosynthesis rates and content of thymosin beta 4 in cell lines.
    Arch Biochem Biophys. 1985 Jul;240(1):236-41 PMID: 2990345
  29. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.
    Nature. 1984 Oct 4-10;311(5985):433-8 PMID: 6090941
  30. Nerve growth factor mediates phosphorylation of specific proteins.
    Cell. 1980 Nov;22(2 Pt 2):571-81 PMID: 6256087
  31. The use of thin acrylamide gels for DNA sequencing.
    FEBS Lett. 1978 Mar 1;87(1):107-10 PMID: 631324
  32. 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):468-76 PMID: 2857777
  33. Nerve growth factor action is mediated by cyclic AMP- and Ca+2/phospholipid-dependent protein kinases.
    J Cell Biol. 1986 Sep;103(3):887-93 PMID: 2875079
  34. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  35. Isolation of developmentally regulated genes from Dictyostelium.
    Dev Biol. 1980 Oct;79(2):409-18 PMID: 7429011
  36. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  37. Superinduction of c-fos by nerve growth factor in the presence of peripherally active benzodiazepines.
    Science. 1985 Sep 20;229(4719):1265-8 PMID: 4035354
  38. 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):2091-5 PMID: 3008151
  39. Choline acetyltransferase activity in striatum of neonatal rats increased by nerve growth factor.
    Science. 1985 Jul 19;229(4710):284-7 PMID: 2861660
  40. 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):1269-73 PMID: 6112744
  41. Molecular cloning of a gene sequence regulated by nerve growth factor.
    Science. 1985 Jul 26;229(4711):393-5 PMID: 3839317
  42. The effect of fibroblast growth factor on PC12 cells.
    J Neurosci. 1985 Feb;5(2):307-16 PMID: 2983039
  43. Sequence of a cloned 523-bp cDNA for thymosin beta 4.
    Arch Biochem Biophys. 1985 Jan;236(1):445-7 PMID: 3838131
  44. 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):2424-8 PMID: 1065897
  45. 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):6059-63 PMID: 310552
  46. 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):787-95 PMID: 6865942
  47. Characterization and isolation of proteolytically modified nerve growth factor.
    Biochemistry. 1976 Dec 14;15(25):5543-52 PMID: 999827
  48. Use of a cloned library for the study of abundant poly(A)+RNA during Xenopus laevis development.
    Dev Biol. 1980 May;76(2):449-64 PMID: 7390013
  49. 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):7537-41 PMID: 3463983
  50. 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):457-65 PMID: 6141171
  51. 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):1162-6 PMID: 6940133
  52. Environmental influences in the development of neural crest derivatives: glucocorticoids, growth factors, and chromaffin cell plasticity.
    J Neurosci. 1985 Aug;5(8):2119-42 PMID: 4020432
  53. 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):1580-4 PMID: 6608728
  54. Stimulation of neuronal acetylcholine receptors induces rapid gene transcription.
    Science. 1986 Oct 3;234(4772):80-3 PMID: 3749894
  55. Trans-synaptic increase in RNA coding for tyrosine hydroxylase in a rat sympathetic ganglion.
    Brain Res. 1985 Jul 22;339(1):151-3 PMID: 2862954
  56. Growth-related changes in specific mRNAs of cultured mouse cells.
    Proc Natl Acad Sci U S A. 1983 Jul;80(14):4271-5 PMID: 6576337
  57. 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):3963-5 PMID: 6575390
  58. The GenBank genetic sequence databank.
    Nucleic Acids Res. 1986 Jan 10;14(1):1-4 PMID: 3945546
  59. 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):4255-9 PMID: 6087314
  60. Selective enzyme induction in a nerve growth factor-responsive pheochromocytoma cell line (PC 12).
    Brain Res. 1978 Oct 6;154(1):186-90 PMID: 81091
  61. 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):3498-502 PMID: 28526
  62. 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):14101-10 PMID: 3877054
  63. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
    Biochemistry. 1979 Nov 27;18(24):5294-9 PMID: 518835
  64. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing.
    J Mol Biol. 1980 Oct 25;143(2):161-78 PMID: 6260957
  65. Thymosin beta 4: a ubiquitous peptide in rat and mouse tissues.
    Proc Natl Acad Sci U S A. 1982 Apr;79(7):2172-5 PMID: 6954532
  66. Nerve growth factor.
    Physiol Rev. 1968 Jul;48(3):534-69 PMID: 4875350
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.7.9.3156-3167.1987
Published
1987-09-00
Pages
3156-67
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367950
Subset
IM
Grants
NIGMS NIH HHS · 5 T32 GM 07308 · United States
NIMH NIH HHS · MH 35976 · United States
NINDS NIH HHS · NS 16036 · United States
External Links
PubMed source DOI: 10.1128/mcb.7.9.3156-3167.1987 PMC Full Text View journal details
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