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
. 1984 Mar;81(6):1867–1870. doi: 10.1073/pnas.81.6.1867

Spiperone: evidence for uptake into secretory granules.

P S Dannies, M S Rudnick, H Fishkes, G Rudnick
PMCID: PMC345023  PMID: 6584920

Abstract

Spiperone, a dopamine antagonist widely used as a specific ligand for dopamine and serotonin receptors, is actively accumulated into the F4C1 strain of rat pituitary tumor cells. The accumulation of 10 nM [3H]spiperone was linear for 3 min and reached a steady state after 10 min. Spiperone accumulation was reduced 50% by preincubation with 5 microM reserpine, an inhibitor of biogenic amine transport into secretory granules, and was also blocked by monensin and ammonium chloride, both of which increase the pH of intracellular storage organelles. Uptake was not affected by replacing sodium in the buffer with lithium at equimolar concentrations. Spiperone at 1 microM inhibited by over 50% serotonin transport into membrane vesicles isolated from platelet dense granules; this concentration inhibited the Na+-dependent plasma membrane transport system less than 10%. The data indicate spiperone specifically interacts with the secretory granule amine transport system and suggest that this transport system is found in the F4C1 pituitary cell strain as well as in platelets and neurons. The data also suggest that experiments utilizing spiperone to measure dopamine and serotonin receptors be interpreted with caution.

Full text

PDF
1867

Selected References

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

  1. Caron M. G., Beaulieu M., Raymond V., Gagné B., Drouin J., Lefkowitz R. J., Labrie F. Dopaminergic receptors in the anterior pituitary gland. Correlation of [3H]dihydroergocryptine binding with the dopaminergic control of prolactin release. J Biol Chem. 1978 Apr 10;253(7):2244–2253. [PubMed] [Google Scholar]
  2. Cronin M. J., Roberts J. M., Weiner R. I. Dopamine and dihydroergocryptine binding to the anterior pituitary and other brain areas of the rat and sheep. Endocrinology. 1978 Jul;103(1):302–309. doi: 10.1210/endo-103-1-302. [DOI] [PubMed] [Google Scholar]
  3. Cronin M. J., Weiner R. I. [3H]Spiroperidol (spiperone) binding to a putative dopamine receptor in sheep and steer pituitary and stalk median eminence. Endocrinology. 1979 Feb;104(2):307–312. doi: 10.1210/endo-104-2-307. [DOI] [PubMed] [Google Scholar]
  4. Fishkes H., Rudnick G. Bioenergetics of serotonin transport by membrane vesicles derived from platelet dense granules. J Biol Chem. 1982 May 25;257(10):5671–5677. [PubMed] [Google Scholar]
  5. Gibbs D. M., Neill J. D. Dopamine levels in hypophysial stalk blood in the rat are sufficient to inhibit prolactin secretion in vivo. Endocrinology. 1978 Jun;102(6):1895–1900. doi: 10.1210/endo-102-6-1895. [DOI] [PubMed] [Google Scholar]
  6. Gudelsky G. A., Nansel D. D., Porter J. C. Uptake and processing of dopamine by cells of the anterior pituitary gland. Endocrinology. 1980 Jul;107(1):30–34. doi: 10.1210/endo-107-1-30. [DOI] [PubMed] [Google Scholar]
  7. Koe B. K. Molecular geometry of inhibitors of the uptake of catecholamines and serotonin in synaptosomal preparations of rat brain. J Pharmacol Exp Ther. 1976 Dec;199(3):649–661. [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. List S. J., Seeman P. Resolution of dopamine and serotonin receptor components of [3H]spiperone binding to rat brain regions. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2620–2624. doi: 10.1073/pnas.78.4.2620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MacLeod R. M., Fontham E. H., Lehmeyer J. E. Prolactin and growth hormone production as influenced by catecholamines and agents that affect brain catecholamines. Neuroendocrinology. 1970;6(5):283–294. doi: 10.1159/000121933. [DOI] [PubMed] [Google Scholar]
  11. Nansel D. D., Gudelsky G. A., Porter J. C. Subcellular localization of dopamine in the anterior pituitary gland of the rat: apparent association of dopamine with prolactin secretory granules. Endocrinology. 1979 Nov;105(5):1073–1077. doi: 10.1210/endo-105-5-1073. [DOI] [PubMed] [Google Scholar]
  12. Richardson U. I. Establishment in culture of a multihormone-secreting cell strain derived from the MtT/F4 rat pituitary tumor. J Cell Physiol. 1976 Jul;88(3):287–296. doi: 10.1002/jcp.1040880304. [DOI] [PubMed] [Google Scholar]
  13. Rudnick G. Active transport of 5-hydroxytryptamine by plasma membrane vesicles isolated from human blood platelets. J Biol Chem. 1977 Apr 10;252(7):2170–2174. [PubMed] [Google Scholar]
  14. Rudnick G., Fishkes H., Nelson P. J., Schuldiner S. Evidence for two distinct serotonin transport systems in platelets. J Biol Chem. 1980 Apr 25;255(8):3638–3641. [PubMed] [Google Scholar]
  15. Shaskan E. G., Snyder S. H. Kinetics of serotonin accumulation into slices from rat brain: relationship to catecholamine uptake. J Pharmacol Exp Ther. 1970 Nov;175(2):404–418. [PubMed] [Google Scholar]
  16. Stitzel R. E. The biological fate of reserpine. Pharmacol Rev. 1976 Sep;28(3):179–208. [PubMed] [Google Scholar]
  17. Tashjian A. H., Jr, Bancroft F. C., Levine L. Production of both prolactin and growth hormone by clonal strains of rat pituitary tumor cells. Differential effects of hydrocortisone and tissue extracts. J Cell Biol. 1970 Oct;47(1):61–70. doi: 10.1083/jcb.47.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Withy R. M., Mayer R. J., Strange P. G. Use of [3H]spiperone for labelling dopaminergic and serotonergic receptors in bovine caudate nucleus. J Neurochem. 1981 Nov;37(5):1144–1154. doi: 10.1111/j.1471-4159.1981.tb04664.x. [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