Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Mar;84(5):1309–1313. doi: 10.1073/pnas.84.5.1309

Cell-type-specific responses of RT4 neural cell lines to dibutyryl-cAMP: branch determination versus maturation.

K Droms, N Sueoka
PMCID: PMC304417  PMID: 3029777

Abstract

This report describes the induction of cell-type-specific maturation, by dibutyryl-cAMP and testololactone, of neuronal and glial properties in a family of cell lines derived from a rat peripheral neurotumor, RT4. This maturation allows further understanding of the process of determination because of the close lineage relationship between the cell types of the RT4 family. The RT4 family is characterized by the spontaneous conversion of one of the cell types, RT4-AC (stem-cell type), to any of three derivative cell types, RT4-B, RT4-D, or RT4-E, with a frequency of about 10(-5). The RT4-AC cells express some properties characteristic of both neuronal and glial cells. Of these neural properties expressed by RT4-AC cells, only the neuronal properties are expressed by the RT4-B and RT4-E cells, and only the glial properties are expressed by the RT4-D cells. This in vitro cell-type conversion of RT4-AC to three derivative cell types is a branch point for the coordinate regulation of several properties and seems to resemble determination in vivo. In our standard culture conditions, several other neuronal and glial properties are not expressed by these cell types. However, addition of dibutyryl-cAMP induces expression of additional properties, in a cell-type-specific manner: formation of long cellular processes in the RT4-B8 and RT4-E5 cell lines and expression of high-affinity uptake of gamma-aminobutyric acid, by a glial-cell-specific mechanism, in the RT4-D6-2 cell line. These new properties are maximally expressed 2-3 days after addition of dibutyryl-cAMP. This indicates that conversion of RT4-AC to the derivative cell types is also a branch point for the regulation of cell-type-specific properties whose expression is responsive to cAMP. Thus, the potential for maturation in response to increased cAMP is a property that segregates in a cell-type-specific manner and is activated at the determinational level in this system.

Full text

PDF
1309

Images in this article

1310Image
on p.1310

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett D. C., Peachey L. A., Durbin H., Rudland P. S. A possible mammary stem cell line. Cell. 1978 Sep;15(1):283–298. doi: 10.1016/0092-8674(78)90104-6. [DOI] [PubMed] [Google Scholar]
  3. Bevan S., Raff M. Voltage-dependent potassium currents in cultured astrocytes. Nature. 1985 May 16;315(6016):229–232. doi: 10.1038/315229a0. [DOI] [PubMed] [Google Scholar]
  4. Bowery N. G., Brown D. A. -Aminobutyric acid uptake by sympathetic ganglia. Nat New Biol. 1972 Jul 19;238(81):89–91. doi: 10.1038/newbio238089a0. [DOI] [PubMed] [Google Scholar]
  5. Chiu S. Y., Schrager P., Ritchie J. M. Neuronal-type Na+ and K+ channels in rabbit cultured Schwann cells. Nature. 1984 Sep 13;311(5982):156–157. doi: 10.1038/311156a0. [DOI] [PubMed] [Google Scholar]
  6. De Vitry F., Picart R., Jacque C., Legault L., Dupouey P., Tixier-Vidal A. Presumptive common precursor for neuronal and glial cell lineages in mouse hypothalamus. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4165–4169. doi: 10.1073/pnas.77.7.4165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dichter M. A., Tischler A. S., Greene L. A. 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. doi: 10.1038/268501a0. [DOI] [PubMed] [Google Scholar]
  8. Frame M. C., Freshney R. I., Vaughan P. F., Graham D. I., Shaw R. Interrelationship between differentiation and malignancy-associated properties in glioma. Br J Cancer. 1984 Mar;49(3):269–280. doi: 10.1038/bjc.1984.44. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Furmanski P., Silverman D. J., Lubin M. Expression of differentiated functions in mouse neuroblastoma mediated by dibutyryl-cyclic adenosine monophosphate. Nature. 1971 Oct 8;233(5319):413–415. doi: 10.1038/233413a0. [DOI] [PubMed] [Google Scholar]
  10. Gardner-Medwin A. R. Neurophysiology. Potential challenge from glia. Nature. 1985 May 16;315(6016):181–181. doi: 10.1038/315181a0. [DOI] [PubMed] [Google Scholar]
  11. Green H., Meuth M. An established pre-adipose cell line and its differentiation in culture. Cell. 1974 Oct;3(2):127–133. doi: 10.1016/0092-8674(74)90116-0. [DOI] [PubMed] [Google Scholar]
  12. Greene L. A., Rein G. Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheo-chromocytoma cells. Brain Res. 1977 Jul 1;129(2):247–263. doi: 10.1016/0006-8993(77)90005-1. [DOI] [PubMed] [Google Scholar]
  13. Imada M., Sueoka N. Clonal sublines of rat neurotumor RT4 and cell differentiation. I. Isolation and characterization of cell lines and cell type conversion. Dev Biol. 1978 Sep;66(1):97–108. doi: 10.1016/0012-1606(78)90276-2. [DOI] [PubMed] [Google Scholar]
  14. Kao F. T., Faik P., Puck T. T. Extension of branching processes from hybrids of brain and Chinese hamster ovary cells. Exp Cell Res. 1979 Aug;122(1):83–91. doi: 10.1016/0014-4827(79)90563-9. [DOI] [PubMed] [Google Scholar]
  15. Kuwano R., Usui H., Maeda T., Fukui T., Yamanari N., Ohtsuka E., Ikehara M., Takahashi Y. Molecular cloning and the complete nucleotide sequence of cDNA to mRNA for S-100 protein of rat brain. Nucleic Acids Res. 1984 Oct 11;12(19):7455–7465. doi: 10.1093/nar/12.19.7455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kyritsis A. P., Tsokos M., Triche T. J., Chader G. J. Retinoblastoma--origin from a primitive neuroectodermal cell? Nature. 1984 Feb 2;307(5950):471–473. doi: 10.1038/307471a0. [DOI] [PubMed] [Google Scholar]
  17. Lewis S. A., Balcarek J. M., Krek V., Shelanski M., Cowan N. J. Sequence of a cDNA clone encoding mouse glial fibrillary acidic protein: structural conservation of intermediate filaments. Proc Natl Acad Sci U S A. 1984 May;81(9):2743–2746. doi: 10.1073/pnas.81.9.2743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. MacVicar B. A. Voltage-dependent calcium channels in glial cells. Science. 1984 Dec 14;226(4680):1345–1347. doi: 10.1126/science.6095454. [DOI] [PubMed] [Google Scholar]
  19. Nirenberg M., Wilson S., Higashida H., Rotter A., Krueger K., Busis N., Ray R., Kenimer J. G., Adler M. Modulation of synapse formation by cyclic adenosine monophosphate. Science. 1983 Nov 18;222(4625):794–799. doi: 10.1126/science.6314503. [DOI] [PubMed] [Google Scholar]
  20. Prasad K. N., Hsie A. W. Morphologic differentiation of mouse neuroblastoma cells induced in vitro by dibutyryl adenosine 3':5'-cyclic monophosphate. Nat New Biol. 1971 Sep 29;233(39):141–142. doi: 10.1038/newbio233141a0. [DOI] [PubMed] [Google Scholar]
  21. Rohrer H., Sommer I. Simultaneous expression of neuronal and glial properties by chick ciliary ganglion cells during development. J Neurosci. 1983 Aug;3(8):1683–1693. doi: 10.1523/JNEUROSCI.03-08-01683.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roisen F. J., Murphy R. A., Braden W. G. Neurite development in vitro. I. The effects of adenosine 3'5'-cyclic monophosphate (cyclic AMP). J Neurobiol. 1972;3(4):347–368. doi: 10.1002/neu.480030408. [DOI] [PubMed] [Google Scholar]
  23. Roisen F. J., Murphy R. A., Pichichero M. E., Braden W. G. Cyclic adenosine monophosphate stimulation of axonal elongation. Science. 1972 Jan 7;175(4017):73–74. doi: 10.1126/science.175.4017.73. [DOI] [PubMed] [Google Scholar]
  24. Schrier B. K., Thompson E. J. On the role of glial cells in the mammalian nervous system. Uptake, excretion, and metabolism of putative neurotransmitters by cultured glial tumor cells. J Biol Chem. 1974 Mar 25;249(6):1769–1780. [PubMed] [Google Scholar]
  25. Schubert D., Heinemann S., Carlisle W., Tarikas H., Kimes B., Patrick J., Steinbach J. H., Culp W., Brandt B. L. Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224–227. doi: 10.1038/249224a0. [DOI] [PubMed] [Google Scholar]
  26. Sobue G., Pleasure D. Schwann cell galactocerebroside induced by derivatives of adenosine 3',5'-monophosphate. Science. 1984 Apr 6;224(4644):72–74. doi: 10.1126/science.6322307. [DOI] [PubMed] [Google Scholar]
  27. Stallcup W. B. The NG2 antigen, a putative lineage marker: immunofluorescent localization in primary cultures of rat brain. Dev Biol. 1981 Apr 15;83(1):154–165. doi: 10.1016/s0012-1606(81)80018-8. [DOI] [PubMed] [Google Scholar]
  28. Tabuchi K., Imada M., Nishimoto A. Effects of cyclic AMP on S-100 protein level in C-6 glioma cells. J Neurol Sci. 1982 Oct;56(1):57–63. doi: 10.1016/0022-510x(82)90060-0. [DOI] [PubMed] [Google Scholar]
  29. Tomozawa Y., Sueoka N. In vitro segregation of different cell lines with neuronal and glial properties from a stem cell line of rat neurotumor RT4. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6305–6309. doi: 10.1073/pnas.75.12.6305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tomozawa Y., Sueoka N., Miyake M. Clonal sublines of rat neurotumor RT4 and cell differentiation. V. Comparison of Na+ influx, Rb+ efflux, and action potential among stem-cell, neuronal, and glial cell types. Dev Biol. 1985 Apr;108(2):503–512. doi: 10.1016/0012-1606(85)90053-3. [DOI] [PubMed] [Google Scholar]
  31. Wilson S. S., Baetge E. E., Stallcup W. B. Antisera specific for cell lines with mixed neuronal and glial properties. Dev Biol. 1981 Apr 15;83(1):146–153. doi: 10.1016/s0012-1606(81)80017-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES