Skip to main content
SpringerLink
Log in

Enhancement of phospholipid hydrolysis in vasopressin-stimulated BHK-21 and H9c2 cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The hydrolysis of phospholipids in vasopressin-stimulated baby hamster kidney (BHK)-21 and H9c2 myoblastic cells was investigated. Phosphatidylcholine and phosphatidylethanolamine in these cells were pulse labelled with [3H]glycerol, [3H]myristate, [3H]choline or [3H]ethanolamine, and chased with the non-labelled precursor until linear turnover rates were obtained. When cells labelled with [3H]glycerol or [3H]myristate were stimulated by vasopressin, no significant decrease in the labelling of phosphatidylcholine was detected, but the labelling of phosphatidic acid was elevated. However, the labellings of phosphatidylethanolamine and its hydrolytic product were not affected by vasopressin stimulation. When the cells were pulse labelled with [3H]-choline, vasopressin stimulation caused a decrease in the labelled phosphatidylcholine with a corresponding increase in the labelled choline. The apparent discrepancy between the two types of labelling might be explained by the recycling of labelled phosphatidic acid back into phosphatidylcholine, thus masking the reduction in the labelled phospholipid during vasopressin stimulation. Alternatively, the labelled choline produced by vasopressin stimulation was released into the medium, thus reducing the recycling of label precursor back into the phospholipid and making the decrease in the labelling of phosphatidylcholine readily detectable. Further studies revealed that vasopressin treatment caused an enhancement of phospholipase D activity in these cells. The presence of substrate-specific phospholipase D isoforms in mammalian tissues led us to postulate that the differential stimulation of phospholipid hydrolysis by vasopressin was caused by the enhancement of a phosphatidylcholine-specific phospholipase D in both BHK-21 and the H9c2 cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BHK-21 cells:

baby hamster kidney-21 cells

References

  1. Dennis EA, Rhee SG, Billah MM, Hannun YA: Role of phospholipases in generating lipid second messengers in signal transduction. FASEB J 5:2068–2077, 1991

    Google Scholar 

  2. Berridge MJ, Irvine RF: Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature 312:315–321, 1984

    Google Scholar 

  3. Nishizuka Y: Studies and perspectives of protein kinase C. Science 233:305–312, 1986

    Google Scholar 

  4. Exton JH: Signalling through phosphatidylcholine breakdown. J Biol Chem 265:1–4, 1990

    Google Scholar 

  5. Billah MM, Anthes JC: The regulation and cellular functions of phosphatidylcholine hydrolysis. Biochem J 269:281–291, 1990

    Google Scholar 

  6. Besterman JM, Duronio V, Cuatrecasas P: Rapid formation of diacylglycerol from phosphatidylcholine. Proc Natl Acad Sci USA 83:6785–6789, 1986

    Google Scholar 

  7. Augert G, Bocckino SB, Blackmore PF, Exton JH: Hormonal stimulation of diacylglycerol formation in hepatocytes. J Biol Chem 264:21689–21698, 1989

    Google Scholar 

  8. Kiss Z, Anderson WB: Phorbol ester stimulates the hydrolysis of phosphatidylethanolamine in leukemic HL-60, NIH 3T3, and baby hamster kidney cells. J Biol Chem 264:1483–1487, 1989

    Google Scholar 

  9. Kiss Z, Anderson WB: ATP stimulates the hydrolysis of phosphatidylethanolamine in NIH 3T3 cells. J Biol Chem 265:7345–7350, 1990

    Google Scholar 

  10. Hii CST, Edwards YS, Murray AW: Phorbol ester-stimulated hydrolysis of phosphatidylcholine and phosphatidylethanolamine by phospholipase D in HeLa cells. J Biol Chem 266:20238–20243, 1991

    Google Scholar 

  11. Rosoff PM, Savage N, Dinarello CA: Interleukin-I stimulates diacylglycerol production in T lymphocytes by a novel mechanism. Cell 54:73–81, 1988

    Google Scholar 

  12. Lacal JC, Moscat J, Aaronson SA: Novel source of 1,2-diacylglycerol elevated in cells transformed by Ha-ras oncogene. Nature 330:269–272, 1987

    Google Scholar 

  13. Price BD, Morris JDH, Marshall CJ, Hall A: Stimulation of phosphatidylcholine hydrolysis, diacylglycerol release, and arachidonic acid production by oncogenic Ras is a consequence of protein kinase C activation. J Biol Chem 264:16638–16643, 1989

    Google Scholar 

  14. Slivka RS, Meier KE, Insel PA: a1-Adrenergic receptors promote phosphatidylcholine hydrolysis in MDCK-D1 cells. J Biol Chem 263:12242–12246, 1988

    Google Scholar 

  15. Muir JG, Murray AW: Bombesin and phorbol ester stimulate phosphatidylcholine hydrolysis by phospholipase C. J Cell Physiol 130:382–391, 1987

    Google Scholar 

  16. Paddon HB, Vance DE: Tetradecanoyl-phorbol acetate stimulates phosphatidylcholine biosynthesis in Hela cells by an increase in the rate of the reaction catalysed by CTP:phosphocholine cytidylyltransferase. Biochim Biophys Acta 620:636–640, 1980

    Google Scholar 

  17. Daniel LW, Waite M, Wykle RL: A novel mechanism of diglyceride formation. J Biol Chem 261:9128–9132, 1986

    Google Scholar 

  18. Liscovitch M, Slack MB, Blusztajn JK, Wurtman RJ: Differential regulation of phosphatidylcholine biosynthesis by 12-O-tetradecanoylphorbol-13-acetate and diacylglycerol in NG108-15 neuroblastoma x glioma hybrid cells. J Biol Chem 262:17487–17491, 1987

    Google Scholar 

  19. Takuwa N, Takuwa Y, Rasmussen H: A tumour promoter, 12-O-tetradecanoylphorbol 13-acetate, increases cellular 1,2-diacylglycerol content through a mechanism other than phosphoinositide hydrolysis in swiss-mouse 3T3 fibroblasts. Biochem J 243:647–653, 1987

    Google Scholar 

  20. Cabot MC, Welsh CJ, Zhang Z-C, Cao HT, Chabbott H, Lebowitz H: Vasopressin, phorboldiesters and serum elicit choline glycerophospholipid hydrolysis and diacylglycerol formation in nontransformed cells: transformed derivatives do not respond. Biochim Biophys Acta 959:46–57, 1988

    Google Scholar 

  21. Martin TW, Feldman DR, Michaelis K: Phosphatidylcholine hydrolysis stimulated by phorbol myristate acetate is mediated principally by phospholipase D in endothelial cells. Biochim Biophys Acta 1053:162–172, 1990

    Google Scholar 

  22. Man AS, Lee E, Choy PC: Biphasic modulation of choline uptake and phosphatidylcholine biosynthesis by vasopressin in rat cardiac myocytes. Lipids 29:15–19, 1994

    Google Scholar 

  23. Tijburg LBM, Schuurmans EAJM, Geelen MJH, van Golde LMG: Effects of vasopressin on the synthesis of phosphatidylethanolamines and phosphatidylcholines by isolated rat hepatocytes. Biochim Biophys Acta 919:49–57, 1987

    Google Scholar 

  24. Troyer DA, Gonzalez OF, Padilla RM, Kreisberg JI: Vasopressin and phorbol ester-stimulated phosphatidylcholine metabolism in mesangial cells. Am J Physiol 262:F185-F191, 1992

    Google Scholar 

  25. Huang C, Cabot MC: Vasopressin-induced polyphosphoinositide and phosphotidylcholine degradation in fibroblasts. J Biol Chem 265:17468–17473, 1990

    Google Scholar 

  26. Welsh CJ, Schmeichel K, Cao H-T, Chabbott H: Vasopressin stimulates phospholipase D activity against phosphatidylcholine in vascular smooth muscle cells. Lipids 25:675–684, 1990

    Google Scholar 

  27. Hui R, Robillard M, Falardeau P: Inhibition of vasopressin-induced formation of diradylglycerols in vascular smooth muscle cells by incorporation of eicosapentaenoic acid in membrane phospholipids. J Hypertens 10:1145–1153, 1992

    Google Scholar 

  28. Zha X, Jay FT, Choy PC: Effects of amino acids and ethanolamine on choline uptake and phosphatidylcholine biosynthesis in baby hamster kidney-21 cells. Biochem Cell Biol 70:1319–1324, 1992

    Google Scholar 

  29. Bligh EG, Dyer WJ: A rapid method of total lipid extraction and purification. Can J Biochem 37:911–917, 1959

    Google Scholar 

  30. Bocokino SB, Blackmore PF, Wilson PB, Exton JH: Phosphatidate accumulation in hormone-treated hepatocytes via a phospholipase D metabolism. J Biol Chem 262:15309–15315, 1987

    Google Scholar 

  31. Billah MM, Eckel S, Mullmann TJ, Egan RW, Siegel MI: Phosphatidylcholine hydrolysis of phospholipase D determines phosphatidate and diglyceride levels in chemotactic peptidestimulated human neutrophils. J Biol Chem 264:17069–17077, 1989

    Google Scholar 

  32. Martin TW, Michaelis K: Bradykinin stimulates phosphodiesteratic cleavage of phosphatidylcholine in cultured endothelial cells. Biochem Biophys Res Commun 157:1271–1279, 1988

    Google Scholar 

  33. Tran K, Proulx PP, Chan AC: Vitamin E suppresses diacylglycerol (DAG) level in thrombin-stimulated endothelial cells through an increase of DAG kinase activity. Biochim Biophys Acta 1212:193–202, 1994

    Google Scholar 

  34. Creba JA, Downes CP, Hawkins PT, Brewster G, Michell RH, Kirk CJ: Rapid breakdown of phosphoinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in rat hepatocytes stimulated by vasopressin and other Ca2+-mobilizing hormones. Biochem J 212:733–747, 1983

    Google Scholar 

  35. Xuan Y-T, Su Y-F, Chang K-J, Watkins WD: A pertussis/cholera toxin sensitive G-protein may mediate vasopressin induced inositol phosphate formation in smooth muscle cell. Biochem Biophys Res Commun 146:898–906, 1987

    Google Scholar 

  36. Catt KJ, Balla T: Phosphoinositide metabolism and hormone action. Annu Rev Med 40:487–509, 1989

    Google Scholar 

  37. Martin TW: Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells. Biochim Biophys Acta 962:282–296, 1988

    Google Scholar 

  38. Martin TW, Mikaelis K: P2-purinergic agonists stimulate phosphodiesteratic cleavage of phosphatidylcholine in endothelial cells. J Biol Chem 264:8847–8856, 1989

    Google Scholar 

  39. Horwitz J: Bradykinin activates a phospholipase D that hydrolyses phosphatidylcholine in PC12 cells. J Neurochem 56:509–517, 1991

    Google Scholar 

  40. Holbrook PG, Pannell LK, Murata Y, Daly JW: Molecular species analysis of a product of phospholipase D activation. J Biol Chem 267:16834–16840, 1992

    Google Scholar 

  41. Taki T, Kanfer JN: Partial purification and properties of a rat brain phospholipase D. J Biol Chem 254:9761–9765, 1979

    Google Scholar 

  42. Wang P, Anthes JC, Siegel MI, Egan RW, Billah MM: Existence of cytosolic phospholipase D. J Biol Chem 266:14877–14880, 1991

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Manitoba, 770 Bannatyne Avenue, R3E OW3, Winnipeg, Manitoba, Canada

    Khai Tran, Xiliang Zha, Monroe Chan & Patrick C. Choy

Authors
  1. Khai Tran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiliang Zha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Monroe Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Patrick C. Choy
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tran, K., Zha, X., Chan, M. et al. Enhancement of phospholipid hydrolysis in vasopressin-stimulated BHK-21 and H9c2 cells. Mol Cell Biochem 151, 69–76 (1995). https://doi.org/10.1007/BF01076898

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01076898

Key words

Search

Navigation