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 Jul;84(14):5078–5081. doi: 10.1073/pnas.84.14.5078

Depolarizing influences regulate preprotachykinin mRNA in sympathetic neurons.

A Roach, J E Adler, I B Black
PMCID: PMC305250  PMID: 3474639

Abstract

We have been studying mechanisms regulating neurotransmitter plasticity in sympathetic neurons. Neurons of the rat superior cervical ganglion (SCG) synthesize multiple putative transmitters, including the peptide substance P (SP). We have now examined steady-state levels of the mRNA encoding preprotachykinin (PPT), the SP precursor. A cloned cDNA probe was used to examine regulation mRNA levels in culture and in vivo. In RNA gel blot experiments, a single band (1.1 kilobases long) was observed in all cases in which an RNA was detected. A low level of PPT mRNA was detected by RNase protection assay in uncultured ganglia, suggesting that the low levels of SP previously observed in the normal ganglion in vivo are synthesized locally. When ganglia were maintained in culture, with consequent denervation, the steady-state level of PPT mRNA increased by 25-fold over the first 24 hr, and the high level was maintained for at least 7 days. RNase protection experiments indicated that the major message in the SCG is the beta-PPT mRNA, encoding both SP and neurokinin A peptide regions. Accumulation of the PPT mRNA in cultured ganglia was sharply inhibited by the depolarizing agent veratridine, and this effect was blocked by tetrodotoxin. Therefore, one form of neuronal plasticity, change in neurotransmitter metabolism, is regulated at least in part by altering steady-state levels of specific mRNA. More generally, extracellular signals may contribute to neuronal plasticity through changes in gene expression.

Full text

PDF
5078

Images in this article

5079Image
on p.5079
5079Image
on p.5079
5080Image
on p.5080

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BORNSTEIN M. B. Reconstituted rattail collagen used as substrate for tissue cultures on coverslips in Maximow slides and roller tubes. Lab Invest. 1958 Mar-Apr;7(2):134–137. [PubMed] [Google Scholar]
  3. Bohn M. C., Kessler J. A., Adler J. E., Markey K., Goldstein M., Black I. B. Simultaneous expression of the SP-peptidergic and noradrenergic phenotypes in rat sympathetic neurons. Brain Res. 1984 Apr 30;298(2):378–381. doi: 10.1016/0006-8993(84)91442-2. [DOI] [PubMed] [Google Scholar]
  4. Bottenstein J. E., Sato G. H. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A. 1979 Jan;76(1):514–517. doi: 10.1073/pnas.76.1.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. HAM R. G. CLONAL GROWTH OF MAMMALIAN CELLS IN A CHEMICALLY DEFINED, SYNTHETIC MEDIUM. Proc Natl Acad Sci U S A. 1965 Feb;53:288–293. doi: 10.1073/pnas.53.2.288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hefti F., Gnahn H., Schwab M. E., Thoenen H. Induction of tyrosine hydroxylase by nerve growth factor and by elevated K+ concentrations in cultures of dissociated sympathetic neurons. J Neurosci. 1982 Nov;2(11):1554–1566. doi: 10.1523/JNEUROSCI.02-11-01554.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Joh T. H., Geghman C., Reis D. Immunochemical demonstration of increased accumulation of tyrosine hydroxylase protein in sympathetic ganglia and adrenal medulla elicited by reserpine. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2767–2771. doi: 10.1073/pnas.70.10.2767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kao C. Y. Tetrodotoxin, saxitoxin and their significance in the study of excitation phenomena. Pharmacol Rev. 1966 Jun;18(2):997–1049. [PubMed] [Google Scholar]
  10. Kessler J. A., Adler J. E., Bell W. O., Black I. B. Substance P and somatostatin metabolism in sympathetic and special sensory ganglia in vitro. Neuroscience. 1983 Jun;9(2):309–318. doi: 10.1016/0306-4522(83)90296-8. [DOI] [PubMed] [Google Scholar]
  11. Kessler J. A., Adler J. E., Bohn M. C., Black I. B. Substance P in principal sympathetic neurons: regulation by impulse activity. Science. 1981 Oct 16;214(4518):335–336. doi: 10.1126/science.6169153. [DOI] [PubMed] [Google Scholar]
  12. Kessler J. A., Black I. B. Regulation of substance P in adult rat sympathetic ganglia. Brain Res. 1982 Feb 18;234(1):182–187. doi: 10.1016/0006-8993(82)90485-1. [DOI] [PubMed] [Google Scholar]
  13. Klarsfeld A., Changeux J. P. Activity regulates the levels of acetylcholine receptor alpha-subunit mRNA in cultured chicken myotubes. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4558–4562. doi: 10.1073/pnas.82.13.4558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Krause J. E., Chirgwin J. M., Carter M. S., Xu Z. S., Hershey A. D. Three rat preprotachykinin mRNAs encode the neuropeptides substance P and neurokinin A. Proc Natl Acad Sci U S A. 1987 Feb;84(3):881–885. doi: 10.1073/pnas.84.3.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LaGamma E. F., White J. D., Adler J. E., Krause J. E., McKelvy J. F., Black I. B. Depolarization regulates adrenal preproenkephalin mRNA. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8252–8255. doi: 10.1073/pnas.82.23.8252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lynch G., Baudry M. The biochemistry of memory: a new and specific hypothesis. Science. 1984 Jun 8;224(4653):1057–1063. doi: 10.1126/science.6144182. [DOI] [PubMed] [Google Scholar]
  17. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nawa H., Hirose T., Takashima H., Inayama S., Nakanishi S. Nucleotide sequences of cloned cDNAs for two types of bovine brain substance P precursor. Nature. 1983 Nov 3;306(5938):32–36. doi: 10.1038/306032a0. [DOI] [PubMed] [Google Scholar]
  19. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wiesel T. N. Postnatal development of the visual cortex and the influence of environment. Nature. 1982 Oct 14;299(5884):583–591. doi: 10.1038/299583a0. [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