Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1993 Feb 1;289(Pt 3):675–679. doi: 10.1042/bj2890675

Generation of inositol phosphates, diacylglycerol and calcium fluxes in myoblasts treated with 1,25-dihydroxyvitamin D3.

S Morelli 1, A R de Boland 1, R L Boland 1
PMCID: PMC1132228  PMID: 8382046

Abstract

We have examined the effects of the seco-steroid hormone 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on membrane phosphoinositide metabolism, protein kinase C (PKC) activation and influx of extracellular Ca2+ in chick-embryo muscle-cell (myoblast) cultures. At physiological concentrations, the hormone induces a rapid (15 s) and transient release of inositol triphosphate (InsP3) and diacylglycerol (DAG). InsP3 release is maximal at 60 s (80% above controls) and then declines. The effects of 1,25(OH)2D3 on InsP3 production exhibited specificity, as 25-hydroxy-vitamin D3 and 24,25-dihydroxy-vitamin D3 did not alter myoblast InsP3 levels. The stimulation of DAG is biphasic, with peaks at 60 s (+105%) and 5 min (+700%). The second phase of DAG release is not associated with changes in InsP3. 1,25(OH)2D3 induces a rapid (within 60 s) accumulation of InsP2, and its effect on InsP is delayed (120 s). The hormone rapidly activates myoblast PKC, with maximal translocation of activity from the cytosol to the cell membrane occurring at 60 s. Myoblast 45Ca uptake significantly increases within 30 s of exposure to 1,25(OH)2D3. The response is time- (0.5-10 min) and dose- (1 pM-10 nM) dependent. The effects of the hormone are mimicked by the Ca(2+)-channel agonist Bay K 8644 and are effectively suppressed by nifedipine and extracellular EGTA. The results suggest that the rapid non-genomic actions of 1,25(OH)2D3 in myoblasts involve second-messenger systems associated with the generation of InsP3 and DAG and regulation of Ca2+ fluxes through voltage-operated channels.

Full text

PDF
675

Selected References

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

  1. Anderson W. B., Estival A., Tapiovaara H., Gopalakrishna R. Altered subcellular distribution of protein kinase C (a phorbol ester receptor). Possible role in tumor promotion and the regulation of cell growth: relationship to changes in adenylate cyclase activity. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1985;19:287–306. [PubMed] [Google Scholar]
  2. Baran D. T., Kelly A. M. Lysophosphatidylinositol: a potential mediator of 1,25-dihydroxyvitamin D-induced increments in hepatocyte cytosolic calcium. Endocrinology. 1988 Mar;122(3):930–934. doi: 10.1210/endo-122-3-930. [DOI] [PubMed] [Google Scholar]
  3. Baran D. T., Sorensen A. M., Honeyman T. W., Ray R., Holick M. F. 1 alpha,25-dihydroxyvitamin D3-induced increments in hepatocyte cytosolic calcium and lysophosphatidylinositol: inhibition by pertussis toxin and 1 beta,25-dihydroxyvitamin D3. J Bone Miner Res. 1990 May;5(5):517–524. doi: 10.1002/jbmr.5650050514. [DOI] [PubMed] [Google Scholar]
  4. Barsony J., Pike J. W., DeLuca H. F., Marx S. J. Immunocytology with microwave-fixed fibroblasts shows 1 alpha,25-dihydroxyvitamin D3-dependent rapid and estrogen-dependent slow reorganization of vitamin D receptors. J Cell Biol. 1990 Dec;111(6 Pt 1):2385–2395. doi: 10.1083/jcb.111.6.2385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benham C. D., Tsien R. W. A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle. Nature. 1987 Jul 16;328(6127):275–278. doi: 10.1038/328275a0. [DOI] [PubMed] [Google Scholar]
  6. Berridge M. J., Dawson R. M., Downes C. P., Heslop J. P., Irvine R. F. Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides. Biochem J. 1983 May 15;212(2):473–482. doi: 10.1042/bj2120473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Berridge M. J. Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem. 1987;56:159–193. doi: 10.1146/annurev.bi.56.070187.001111. [DOI] [PubMed] [Google Scholar]
  8. Besterman J. M., Duronio V., Cuatrecasas P. Rapid formation of diacylglycerol from phosphatidylcholine: a pathway for generation of a second messenger. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6785–6789. doi: 10.1073/pnas.83.18.6785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bourdeau A., Atmani F., Grosse B., Lieberherr M. Rapid effects of 1,25-dihydroxyvitamin D3 and extracellular Ca2+ on phospholipid metabolism in dispersed porcine parathyroid cells. Endocrinology. 1990 Dec;127(6):2738–2743. doi: 10.1210/endo-127-6-2738. [DOI] [PubMed] [Google Scholar]
  10. Caffrey J. M., Farach-Carson M. C. Vitamin D3 metabolites modulate dihydropyridine-sensitive calcium currents in clonal rat osteosarcoma cells. J Biol Chem. 1989 Dec 5;264(34):20265–20274. [PubMed] [Google Scholar]
  11. Chiu R., Imagawa M., Imbra R. J., Bockoven J. R., Karin M. Multiple cis- and trans-acting elements mediate the transcriptional response to phorbol esters. Nature. 1987 Oct 15;329(6140):648–651. doi: 10.1038/329648a0. [DOI] [PubMed] [Google Scholar]
  12. Civitelli R., Kim Y. S., Gunsten S. L., Fujimori A., Huskey M., Avioli L. V., Hruska K. A. Nongenomic activation of the calcium message system by vitamin D metabolites in osteoblast-like cells. Endocrinology. 1990 Nov;127(5):2253–2262. doi: 10.1210/endo-127-5-2253. [DOI] [PubMed] [Google Scholar]
  13. Drittanti L. N., Boland R. L., de Boland A. R. Induction of specific proteins in cultured skeletal muscle cells by 1,25-dihydroxyvitamin D-3. Biochim Biophys Acta. 1989 Jun 15;1012(1):16–23. doi: 10.1016/0167-4889(89)90005-0. [DOI] [PubMed] [Google Scholar]
  14. Drittanti L., de Boland A. R., Boland R. Stimulation of calmodulin synthesis in proliferating myoblasts by 1,25-dihydroxy-vitamin D3. Mol Cell Endocrinol. 1990 Dec 3;74(2):143–153. doi: 10.1016/0303-7207(90)90116-p. [DOI] [PubMed] [Google Scholar]
  15. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  16. Fernandez L. M., Massheimer V., de Boland A. R. Cyclic AMP-dependent membrane protein phosphorylation and calmodulin binding are involved in the rapid stimulation of muscle calcium uptake by 1,25-dihydroxyvitamin D3. Calcif Tissue Int. 1990 Nov;47(5):314–319. doi: 10.1007/BF02555915. [DOI] [PubMed] [Google Scholar]
  17. Giuliani D. L., Boland R. L. Effects of vitamin D3 metabolites on calcium fluxes in intact chicken skeletal muscle and myoblasts cultured in vitro. Calcif Tissue Int. 1984 Mar;36(2):200–205. doi: 10.1007/BF02405318. [DOI] [PubMed] [Google Scholar]
  18. Kano M., Wakuta K., Satoh R. Two components of calcium channel current in embryonic chick skeletal muscle cells developing in culture. Brain Res Dev Brain Res. 1989 May 1;47(1):101–112. doi: 10.1016/0165-3806(89)90112-0. [DOI] [PubMed] [Google Scholar]
  19. Kikkawa U., Minakuchi R., Takai Y., Nishizuka Y. Calcium-activated, phospholipid-dependent protein kinase (protein kinase C) from rat brain. Methods Enzymol. 1983;99:288–298. doi: 10.1016/0076-6879(83)99064-x. [DOI] [PubMed] [Google Scholar]
  20. Kuno M., Gardner P. Ion channels activated by inositol 1,4,5-trisphosphate in plasma membrane of human T-lymphocytes. Nature. 1987 Mar 19;326(6110):301–304. doi: 10.1038/326301a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Lacal J. C., Moscat J., Aaronson S. A. Novel source of 1,2-diacylglycerol elevated in cells transformed by Ha-ras oncogene. Nature. 1987 Nov 19;330(6145):269–272. doi: 10.1038/330269a0. [DOI] [PubMed] [Google Scholar]
  23. Lieberherr M. Effects of vitamin D3 metabolites on cytosolic free calcium in confluent mouse osteoblasts. J Biol Chem. 1987 Sep 25;262(27):13168–13173. doi: 10.1515/9783110846713.769. [DOI] [PubMed] [Google Scholar]
  24. Lieberherr M., Grosse B., Duchambon P., Drüeke T. A functional cell surface type receptor is required for the early action of 1,25-dihydroxyvitamin D3 on the phosphoinositide metabolism in rat enterocytes. J Biol Chem. 1989 Dec 5;264(34):20403–20406. [PubMed] [Google Scholar]
  25. Lückhoff A., Clapham D. E. Inositol 1,3,4,5-tetrakisphosphate activates an endothelial Ca(2+)-permeable channel. Nature. 1992 Jan 23;355(6358):356–358. doi: 10.1038/355356a0. [DOI] [PubMed] [Google Scholar]
  26. Massheimer V., de Boland A. R. Modulation of 1,25-dihydroxyvitamin D3-dependent Ca2+ uptake in skeletal muscle by protein kinase C. Biochem J. 1992 Jan 15;281(Pt 2):349–352. doi: 10.1042/bj2810349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Neill M. C., Stockdale F. E. A kinetic analysis of myogenesis in vitro. J Cell Biol. 1972 Jan;52(1):52–65. doi: 10.1083/jcb.52.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Oshima J., Watanabe M., Hirosumi J., Orimo H. 1,25(OH)2D3 increases cytosolic Ca++ concentration of osteoblastic cells, clone MC3T3-E1. Biochem Biophys Res Commun. 1987 Jun 15;145(2):956–960. doi: 10.1016/0006-291x(87)91058-8. [DOI] [PubMed] [Google Scholar]
  29. Selles J., Boland R. Evidence on the participation of the 3',5'-cyclic AMP pathway in the non-genomic action of 1,25-dihydroxy-vitamin D3 in cardiac muscle. Mol Cell Endocrinol. 1991 Dec;82(2-3):229–235. doi: 10.1016/0303-7207(91)90036-r. [DOI] [PubMed] [Google Scholar]
  30. Selles J., Boland R. Rapid stimulation of calcium uptake and protein phosphorylation in isolated cardiac muscle by 1,25-dihydroxyvitamin D3. Mol Cell Endocrinol. 1991 May;77(1-3):67–73. doi: 10.1016/0303-7207(91)90059-2. [DOI] [PubMed] [Google Scholar]
  31. Tang W., Ziboh V. A., Isseroff R. R., Martinez D. Novel regulatory actions of 1 alpha,25-dihydroxyvitamin D3 on the metabolism of polyphosphoinositides in murine epidermal keratinocytes. J Cell Physiol. 1987 Jul;132(1):131–136. doi: 10.1002/jcp.1041320118. [DOI] [PubMed] [Google Scholar]
  32. Tornquist K., Tashjian A. H., Jr Dual actions of 1,25-dihydroxycholecalciferol on intracellular Ca2+ in GH4C1 cells: evidence for effects on voltage-operated Ca2+ channels and Na+/Ca2+ exchange. Endocrinology. 1989 Jun;124(6):2765–2776. doi: 10.1210/endo-124-6-2765. [DOI] [PubMed] [Google Scholar]
  33. Vesely D. L., Juan D. Cation-dependent vitamin D activation of human renal cortical guanylate cyclase. Am J Physiol. 1984 Jan;246(1 Pt 1):E115–E120. doi: 10.1152/ajpendo.1984.246.1.E115. [DOI] [PubMed] [Google Scholar]
  34. Wali R. K., Baum C. L., Sitrin M. D., Brasitus T. A. 1,25(OH)2 vitamin D3 stimulates membrane phosphoinositide turnover, activates protein kinase C, and increases cytosolic calcium in rat colonic epithelium. J Clin Invest. 1990 Apr;85(4):1296–1303. doi: 10.1172/JCI114567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ways D. K., Dodd R. C., Bennett T. E., Gray T. K., Earp H. S. 1,25-Dihydroxyvitamin D3 enhances phorbol ester-stimulated differentiation and protein kinase C-dependent substrate phosphorylation activity in the U937 human monoblastoid cell. Endocrinology. 1987 Nov;121(5):1654–1661. doi: 10.1210/endo-121-5-1654. [DOI] [PubMed] [Google Scholar]
  36. de Boland A. R., Boland R. L. Rapid changes in skeletal muscle calcium uptake induced in vitro by 1,25-dihydroxyvitamin D3 are suppressed by calcium channel blockers. Endocrinology. 1987 May;120(5):1858–1864. doi: 10.1210/endo-120-5-1858. [DOI] [PubMed] [Google Scholar]
  37. de Boland A. R., Nemere I., Norman A. W. Ca2(+)-channel agonist BAY K8644 mimics 1,25(OH)2-vitamin D3 rapid enhancement of Ca2+ transport in chick perfused duodenum. Biochem Biophys Res Commun. 1990 Jan 15;166(1):217–222. doi: 10.1016/0006-291x(90)91933-j. [DOI] [PubMed] [Google Scholar]
  38. de Boland A. R., Norman A. W. Influx of extracellular calcium mediates 1,25-dihydroxyvitamin D3-dependent transcaltachia (the rapid stimulation of duodenal Ca2+ transport). Endocrinology. 1990 Nov;127(5):2475–2480. doi: 10.1210/endo-127-5-2475. [DOI] [PubMed] [Google Scholar]
  39. de Boland A. R., Norman A. Evidence for involvement of protein kinase C and cyclic adenosine 3',5' monophosphate-dependent protein kinase in the 1,25-dihydroxy-vitamin D3-mediated rapid stimulation of intestinal calcium transport, (transcaltachia). Endocrinology. 1990 Jul;127(1):39–45. doi: 10.1210/endo-127-1-39. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES