Skip to main content
NCBI home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1991 Sep;104(1):202–206. doi: 10.1111/j.1476-5381.1991.tb12408.x

Inositol 1,4,5-triphosphate-stimulated calcium release from permeabilized cerebellar granule cells.

E M Whitham 1, R A Challiss 1, S R Nahorski 1
PMCID: PMC1908263  PMID: 1786511

Abstract

1. Muscarinic cholinoceptor stimulation of phosphoinositide hydrolysis in rat cultured cerebellar granule cells results in a rapid, transient accumulation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), which has been implicated in the release of non-mitochondrial intracellular Ca2+ stores. In the present study, the release of Ca2+ from intracellular stores and the Ins(1,4,5)P3 receptor responsible for this process have been investigated. 2. Monolayers of saponin-permeabilized granule cells accumulate 45Ca2+ in an ATP-dependent manner and the sequestered 45Ca2+ can be concentration-dependently released by Ins(1,4,5)P3 by a stereospecific and heparin-sensitive mechanism. The EC50 for Ins(1,4,5)P3-stimulated 45Ca2+ release was 80 +/- 3 nM. 3. Radioligand binding studies performed on a crude granule cell membrane fraction indicated the presence of an apparently homogeneous population of stereo-specific Ins(1,4,5)P3 receptors (KD 54.7 +/- 2.0 nM; Bmax 1.37 +/- 0.29 pmol mg-1 protein). 4. This study provides evidence for Ins(1,4,5)P3-sensitive intracellular Ca2+ stores in primary cultures of cerebellar granule cells and suggest that these cells provide an excellent model neuronal system in which to study the relative functional roles of Ca2+ release from intracellular stores and Ca(2+)-entry in neuronal Ca2+ homeostasis.

Full text

PDF
202

Selected References

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

  1. Ambler S. K., Taylor P. Release of nonmitochondrial sequestered Ca2+ from permeabilized muscle cells in culture. Mol Pharmacol. 1989 Mar;35(3):369–374. [PubMed] [Google Scholar]
  2. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  3. Bouchelouche P., Belhage B., Frandsen A., Drejer J., Schousboe A. Glutamate receptor activation in cultured cerebellar granule cells increases cytosolic free Ca2+ by mobilization of cellular Ca2+ and activation of Ca2+ influx. Exp Brain Res. 1989;76(2):281–291. doi: 10.1007/BF00247888. [DOI] [PubMed] [Google Scholar]
  4. Burgoyne R. D., Cambray-Deakin M. A. The cellular neurobiology of neuronal development: the cerebellar granule cell. Brain Res. 1988 Jan-Mar;472(1):77–101. doi: 10.1016/0165-0173(88)90006-9. [DOI] [PubMed] [Google Scholar]
  5. Burgoyne R. D., Pearce I. A., Cambray-Deakin M. N-methyl-D-aspartate raises cytosolic calcium concentration in rat cerebellar granule cells in culture. Neurosci Lett. 1988 Aug 15;91(1):47–52. doi: 10.1016/0304-3940(88)90247-9. [DOI] [PubMed] [Google Scholar]
  6. Carboni E., Wojcik W. J. Dihydropyridine binding sites regulate calcium influx through specific voltage-sensitive calcium channels in cerebellar granule cells. J Neurochem. 1988 Apr;50(4):1279–1286. doi: 10.1111/j.1471-4159.1988.tb10605.x. [DOI] [PubMed] [Google Scholar]
  7. Challiss R. A., Chilvers E. R., Willcocks A. L., Nahorski S. R. Heterogeneity of [3H]inositol 1,4,5-trisphosphate binding sites in adrenal-cortical membranes. Characterization and validation of a radioreceptor assay. Biochem J. 1990 Jan 15;265(2):421–427. doi: 10.1042/bj2650421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheek T. R., O'Sullivan A. J., Moreton R. B., Berridge M. J., Burgoyne R. D. Spatial localization of the stimulus-induced rise in cytosolic Ca2+ in bovine adrenal chromaffin cells. Distinct nicotinic and muscarinic patterns. FEBS Lett. 1989 Apr 24;247(2):429–434. doi: 10.1016/0014-5793(89)81385-7. [DOI] [PubMed] [Google Scholar]
  9. Chuang D. M. Neurotransmitter receptors and phosphoinositide turnover. Annu Rev Pharmacol Toxicol. 1989;29:71–110. doi: 10.1146/annurev.pa.29.040189.000443. [DOI] [PubMed] [Google Scholar]
  10. Ciardo A., Meldolesi J. Regulation of intracellular calcium in cerebellar granule neurons: effects of depolarization and of glutamatergic and cholinergic stimulation. J Neurochem. 1991 Jan;56(1):184–191. doi: 10.1111/j.1471-4159.1991.tb02579.x. [DOI] [PubMed] [Google Scholar]
  11. Courtney M. J., Lambert J. J., Nicholls D. G. The interactions between plasma membrane depolarization and glutamate receptor activation in the regulation of cytoplasmic free calcium in cultured cerebellar granule cells. J Neurosci. 1990 Dec;10(12):3873–3879. doi: 10.1523/JNEUROSCI.10-12-03873.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DeLean A., Munson P. J., Rodbard D. Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Am J Physiol. 1978 Aug;235(2):E97–102. doi: 10.1152/ajpendo.1978.235.2.E97. [DOI] [PubMed] [Google Scholar]
  13. Furuichi T., Shiota C., Mikoshiba K. Distribution of inositol 1,4,5-trisphosphate receptor mRNA in mouse tissues. FEBS Lett. 1990 Jul 2;267(1):85–88. doi: 10.1016/0014-5793(90)80294-s. [DOI] [PubMed] [Google Scholar]
  14. Furuichi T., Yoshikawa S., Miyawaki A., Wada K., Maeda N., Mikoshiba K. Primary structure and functional expression of the inositol 1,4,5-trisphosphate-binding protein P400. Nature. 1989 Nov 2;342(6245):32–38. doi: 10.1038/342032a0. [DOI] [PubMed] [Google Scholar]
  15. Gallo V., Ciotti M. T., Coletti A., Aloisi F., Levi G. Selective release of glutamate from cerebellar granule cells differentiating in culture. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7919–7923. doi: 10.1073/pnas.79.24.7919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gallo V., Giovannini C., Levi G. Modulation of non-N-methyl-D-aspartate receptors in cultured cerebellar granule cells. J Neurochem. 1990 May;54(5):1619–1625. doi: 10.1111/j.1471-4159.1990.tb01213.x. [DOI] [PubMed] [Google Scholar]
  17. Guillemette G., Lamontagne S., Boulay G., Mouillac B. Differential effects of heparin on inositol 1,4,5-trisphosphate binding, metabolism, and calcium release activity in the bovine adrenal cortex. Mol Pharmacol. 1989 Mar;35(3):339–344. [PubMed] [Google Scholar]
  18. Holopainen I., Enkvist M. O., Akerman K. E. Glutamate receptor agonists increase intracellular Ca2+ independently of voltage-gated Ca2+ channels in rat cerebellar granule cells. Neurosci Lett. 1989 Mar 13;98(1):57–62. doi: 10.1016/0304-3940(89)90373-x. [DOI] [PubMed] [Google Scholar]
  19. Hynie S., Wroblewski J. T., Costa E. Profile of phosphatidylinositol metabolism stimulated by carbachol and glutamate in primary cultures of rat cerebellar neurons. Neuropharmacology. 1989 Dec;28(12):1309–1315. doi: 10.1016/0028-3908(89)90004-x. [DOI] [PubMed] [Google Scholar]
  20. Irving A. J., Schofield J. G., Watkins J. C., Sunter D. C., Collingridge G. L. 1S,3R-ACPD stimulates and L-AP3 blocks Ca2+ mobilization in rat cerebellar neurons. Eur J Pharmacol. 1990 Sep 21;186(2-3):363–365. doi: 10.1016/0014-2999(90)90462-f. [DOI] [PubMed] [Google Scholar]
  21. Joseph S. K., Rice H. L. The relationship between inositol trisphosphate receptor density and calcium release in brain microsomes. Mol Pharmacol. 1989 Mar;35(3):355–359. [PubMed] [Google Scholar]
  22. Joseph S. K., Rice H. L., Williamson J. R. The effect of external calcium and pH on inositol trisphosphate-mediated calcium release from cerebellum microsomal fractions. Biochem J. 1989 Feb 15;258(1):261–265. doi: 10.1042/bj2580261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kingsbury A. E., Gallo V., Woodhams P. L., Balazs R. Survival, morphology and adhesion properties of cerebellar interneurones cultured in chemically defined and serum-supplemented medium. Brain Res. 1985 Jan;349(1-2):17–25. doi: 10.1016/0165-3806(85)90128-2. [DOI] [PubMed] [Google Scholar]
  24. Kingsbury A., Balazs R. Effect of calcium agonists and antagonists on cerebellar granule cells. Eur J Pharmacol. 1987 Aug 21;140(3):275–283. doi: 10.1016/0014-2999(87)90284-6. [DOI] [PubMed] [Google Scholar]
  25. Levi G., Patrizio M., Gallo V. Release of endogenous and newly synthesized glutamate and of other amino acids induced by non-N-methyl-D-aspartate receptor activation in cerebellar granule cell cultures. J Neurochem. 1991 Jan;56(1):199–206. doi: 10.1111/j.1471-4159.1991.tb02581.x. [DOI] [PubMed] [Google Scholar]
  26. Lin W. W., Lee C. Y., Chuang D. M. Comparative studies of phosphoinositide hydrolysis induced by endothelin-related peptides in cultured cerebellar astrocytes, C6-glioma and cerebellar granule cells. Biochem Biophys Res Commun. 1990 Apr 30;168(2):512–519. doi: 10.1016/0006-291x(90)92351-y. [DOI] [PubMed] [Google Scholar]
  27. Lin W. W., Lee C. Y., Chuang D. M. Endothelin-1 stimulates the release of preloaded [3H]D-aspartate from cultured cerebellar granule cells. Biochem Biophys Res Commun. 1990 Mar 16;167(2):593–599. doi: 10.1016/0006-291x(90)92066-9. [DOI] [PubMed] [Google Scholar]
  28. Maeda N., Niinobe M., Mikoshiba K. A cerebellar Purkinje cell marker P400 protein is an inositol 1,4,5-trisphosphate (InsP3) receptor protein. Purification and characterization of InsP3 receptor complex. EMBO J. 1990 Jan;9(1):61–67. doi: 10.1002/j.1460-2075.1990.tb08080.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mignery G. A., Südhof T. C., Takei K., De Camilli P. Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor. Nature. 1989 Nov 9;342(6246):192–195. doi: 10.1038/342192a0. [DOI] [PubMed] [Google Scholar]
  30. Nahorski S. R. Inositol polyphosphates and neuronal calcium homeostasis. Trends Neurosci. 1988 Oct;11(10):444–448. doi: 10.1016/0166-2236(88)90196-8. [DOI] [PubMed] [Google Scholar]
  31. Nahorski S. R., Potter B. V. Molecular recognition of inositol polyphosphates by intracellular receptors and metabolic enzymes. Trends Pharmacol Sci. 1989 Apr;10(4):139–144. doi: 10.1016/0165-6147(89)90165-x. [DOI] [PubMed] [Google Scholar]
  32. Nicoletti F., Wroblewski J. T., Novelli A., Alho H., Guidotti A., Costa E. The activation of inositol phospholipid metabolism as a signal-transducing system for excitatory amino acids in primary cultures of cerebellar granule cells. J Neurosci. 1986 Jul;6(7):1905–1911. doi: 10.1523/JNEUROSCI.06-07-01905.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. O'Sullivan A. J., Cheek T. R., Moreton R. B., Berridge M. J., Burgoyne R. D. Localization and heterogeneity of agonist-induced changes in cytosolic calcium concentration in single bovine adrenal chromaffin cells from video imaging of fura-2. EMBO J. 1989 Feb;8(2):401–411. doi: 10.1002/j.1460-2075.1989.tb03391.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ross C. A., Meldolesi J., Milner T. A., Satoh T., Supattapone S., Snyder S. H. Inositol 1,4,5-trisphosphate receptor localized to endoplasmic reticulum in cerebellar Purkinje neurons. Nature. 1989 Jun 8;339(6224):468–470. doi: 10.1038/339468a0. [DOI] [PubMed] [Google Scholar]
  35. Stauderman K. A., Harris G. D., Lovenberg W. Characterization of inositol 1,4,5-trisphosphate-stimulated calcium release from rat cerebellar microsomal fractions. Comparison with [3H]inositol 1,4,5-trisphosphate binding. Biochem J. 1988 Oct 15;255(2):677–683. [PMC free article] [PubMed] [Google Scholar]
  36. Strupish J., Cooke A. M., Potter B. V., Gigg R., Nahorski S. R. Stereospecific mobilization of intracellular Ca2+ by inositol 1,4,5-triphosphate. Comparison with inositol 1,4,5-trisphosphorothioate and inositol 1,3,4-trisphosphate. Biochem J. 1988 Aug 1;253(3):901–905. doi: 10.1042/bj2530901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Supattapone S., Worley P. F., Baraban J. M., Snyder S. H. Solubilization, purification, and characterization of an inositol trisphosphate receptor. J Biol Chem. 1988 Jan 25;263(3):1530–1534. [PubMed] [Google Scholar]
  38. Thayer S. A., Perney T. M., Miller R. J. Regulation of calcium homeostasis in sensory neurons by bradykinin. J Neurosci. 1988 Nov;8(11):4089–4097. doi: 10.1523/JNEUROSCI.08-11-04089.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Whitham E. M., Challiss R. A., Nahorski S. R. M3 muscarinic cholinoceptors are linked to phosphoinositide metabolism in rat cerebellar granule cells. Eur J Pharmacol. 1991 Mar 25;206(3):181–189. doi: 10.1016/s0922-4106(05)80017-3. [DOI] [PubMed] [Google Scholar]
  40. Willcocks A. L., Cooke A. M., Potter B. V., Nahorski S. R. Stereospecific recognition sites for [3H]inositol(1,4,5)-triphosphate in particulate preparations of rat cerebellum. Biochem Biophys Res Commun. 1987 Aug 14;146(3):1071–1078. doi: 10.1016/0006-291x(87)90756-x. [DOI] [PubMed] [Google Scholar]
  41. Worley P. F., Baraban J. M., Colvin J. S., Snyder S. H. Inositol trisphosphate receptor localization in brain: variable stoichiometry with protein kinase C. Nature. 1987 Jan 8;325(7000):159–161. doi: 10.1038/325159a0. [DOI] [PubMed] [Google Scholar]
  42. Worley P. F., Baraban J. M., Snyder S. H. Inositol 1,4,5-trisphosphate receptor binding: autoradiographic localization in rat brain. J Neurosci. 1989 Jan;9(1):339–346. doi: 10.1523/JNEUROSCI.09-01-00339.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Worley P. F., Baraban J. M., Supattapone S., Wilson V. S., Snyder S. H. Characterization of inositol trisphosphate receptor binding in brain. Regulation by pH and calcium. J Biol Chem. 1987 Sep 5;262(25):12132–12136. [PubMed] [Google Scholar]
  44. Wroblewski J. T., Nicoletti F., Costa E. Different coupling of excitatory amino acid receptors with Ca2+ channels in primary cultures of cerebellar granule cells. Neuropharmacology. 1985 Sep;24(9):919–921. doi: 10.1016/0028-3908(85)90046-2. [DOI] [PubMed] [Google Scholar]
  45. Xu J., Chuang D. M. Serotonergic, adrenergic and histaminergic receptors coupled to phospholipase C in cultured cerebellar granule cells of rats. Biochem Pharmacol. 1987 Jul 15;36(14):2353–2358. doi: 10.1016/0006-2952(87)90603-4. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES