Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2003 Nov 1;375(Pt 3):705–712. doi: 10.1042/BJ20021907

Free fatty acids induce cell differentiation to infective forms in Trypanosoma cruzi.

Marisa J Wainszelbaum 1, María L Belaunzarán 1, Estela M Lammel 1, Mónica Florin-Christensen 1, Jorge Florin-Christensen 1, Elvira L D Isola 1
PMCID: PMC1223715  PMID: 12887332

Abstract

Intestinal extracts of Triatoma infestans induce cell differentiation of Trypanosoma cruzi epimastigotes into the infective metacyclic form. Part of this effect can be explained by the presence of haemoglobin fragments, which stimulate trypanosomal adenylate cyclase. In this work we examined the metacyclogenic activity of lipids present in this intestinal extract. We found that lipid extracts of the intestinal extract have significant stimulatory effects that reside with the free-fatty-acid fraction, especially oleic acid. These compounds stimulate de novo diacylglycerol formation and protein kinase C activity in the parasite. Moreover, metacyclogenesis is stimulated by phorbol esters and cell-permeant diacylglycerol, while protein kinase C down-regulation or incubation with inhibitors of this kinase abrogates this effect. These results indicate that free fatty acids are a novel signal, inducing metacyclogenesis, acting through a pathway involving diacylglycerol biosynthesis and protein kinase C activation.

Full Text

The Full Text of this article is available as a PDF (206.0 KB).

Selected References

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

  1. Aihara H., Asaoka Y., Yoshida K., Nishizuka Y. Sustained activation of protein kinase C is essential to HL-60 cell differentiation to macrophage. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11062–11066. doi: 10.1073/pnas.88.24.11062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Augé N., Santanam N., Mori N., Keshava C., Parthasarathy S. Uptake of 13-hydroperoxylinoleic acid by cultured cells. Arterioscler Thromb Vasc Biol. 1999 Apr;19(4):925–931. doi: 10.1161/01.atv.19.4.925. [DOI] [PubMed] [Google Scholar]
  3. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  4. Baron C. B., Cunningham M., Strauss J. F., 3rd, Coburn R. F. Pharmacomechanical coupling in smooth muscle may involve phosphatidylinositol metabolism. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6899–6903. doi: 10.1073/pnas.81.21.6899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brattsand G., Friedrich B., Gullberg M. A rapid separation procedure to analyse phosphorylation of two putative protein kinase C substrates in intact lymphocytes. Scand J Immunol. 1989 Aug;30(2):233–240. doi: 10.1111/j.1365-3083.1989.tb01206.x. [DOI] [PubMed] [Google Scholar]
  6. Brinkmann Joep F. F., Abumrad Nada A., Ibrahimi Azeddine, van der Vusse Ger J., Glatz Jan F. C. New insights into long-chain fatty acid uptake by heart muscle: a crucial role for fatty acid translocase/CD36. Biochem J. 2002 Nov 1;367(Pt 3):561–570. doi: 10.1042/BJ20020747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Contreras V. T., Morel C. M., Goldenberg S. Stage specific gene expression precedes morphological changes during Trypanosoma cruzi metacyclogenesis. Mol Biochem Parasitol. 1985 Jan;14(1):83–96. doi: 10.1016/0166-6851(85)90108-2. [DOI] [PubMed] [Google Scholar]
  8. Crowley M. R., Fineman J. R., Soifer S. J. HA1004, an intracellular calcium antagonist, selectively attenuates pulmonary hypertension in newborn lambs. J Cardiovasc Pharmacol. 1994 May;23(5):806–813. doi: 10.1097/00005344-199405000-00017. [DOI] [PubMed] [Google Scholar]
  9. Denning M. F., Dlugosz A. A., Howett M. K., Yuspa S. H. Expression of an oncogenic rasHa gene in murine keratinocytes induces tyrosine phosphorylation and reduced activity of protein kinase C delta. J Biol Chem. 1993 Dec 15;268(35):26079–26081. [PubMed] [Google Scholar]
  10. Farese R. V., Davis J. S., Barnes D. E., Standaert M. L., Babischkin J. S., Hock R., Rosic N. K., Pollet R. J. The de novo phospholipid effect of insulin is associated with increases in diacylglycerol, but not inositol phosphates or cytosolic Ca2+. Biochem J. 1985 Oct 15;231(2):269–278. doi: 10.1042/bj2310269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farese R. V. Insulin-sensitive phospholipid signaling systems and glucose transport. Update II. Exp Biol Med (Maywood) 2001 Apr;226(4):283–295. doi: 10.1177/153537020122600404. [DOI] [PubMed] [Google Scholar]
  12. Farese R. V., Konda T. S., Davis J. S., Standaert M. L., Pollet R. J., Cooper D. R. Insulin rapidly increases diacylglycerol by activating de novo phosphatidic acid synthesis. Science. 1987 May 1;236(4801):586–589. doi: 10.1126/science.3107122. [DOI] [PubMed] [Google Scholar]
  13. Florin-Christensen J., Florin-Christensen M., Delfino J. M., Rasmussen H. New patterns of diacylglycerol metabolism in intact cells. Biochem J. 1993 Feb 1;289(Pt 3):783–788. doi: 10.1042/bj2890783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Florin-Christensen J., Florin-Christensen M., Delfino J. M., Stegmann T., Rasmussen H. Metabolic fate of plasma membrane diacylglycerols in NIH 3T3 fibroblasts. J Biol Chem. 1992 Jul 25;267(21):14783–14789. [PubMed] [Google Scholar]
  15. Florin-Christensen J., Florin-Christensen M., Meinardi E., Calle R. Diversity of roles of protein kinase C alpha in the proliferation of Swiss 3T3 cells. Biochem J. 1996 Apr 15;315(Pt 2):513–516. doi: 10.1042/bj3150513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Florin-Christensen M., Florin-Christensen J., de Isola E. D., Lammel E., Meinardi E., Brenner R. R., Rasmussen L. Temperature acclimation of Trypanosoma cruzi epimastigote and metacyclic trypomastigote lipids. Mol Biochem Parasitol. 1997 Sep;88(1-2):25–33. doi: 10.1016/s0166-6851(97)00056-x. [DOI] [PubMed] [Google Scholar]
  17. Fraidenraich D., Peña C., Isola E. L., Lammel E. M., Coso O., Añel A. D., Pongor S., Baralle F., Torres H. N., Flawia M. M. Stimulation of Trypanosoma cruzi adenylyl cyclase by an alpha D-globin fragment from Triatoma hindgut: effect on differentiation of epimastigote to trypomastigote forms. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10140–10144. doi: 10.1073/pnas.90.21.10140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Glatz J. F., Storch J. Unravelling the significance of cellular fatty acid-binding proteins. Curr Opin Lipidol. 2001 Jun;12(3):267–274. doi: 10.1097/00041433-200106000-00005. [DOI] [PubMed] [Google Scholar]
  19. González Cappa S. M., Bijovsky A. T., Freilij H., Muller L., Katzin A. M. Aislamiento de una cepa de trypanosoma cruzi a predominio de formas delgadas en la Argentina. Medicina (B Aires) 1981;41(1):119–120. [PubMed] [Google Scholar]
  20. Goodall H., Johnson M. H. Use of carboxyfluorescein diacetate to study formation of permeable channels between mouse blastomeres. Nature. 1982 Feb 11;295(5849):524–526. doi: 10.1038/295524a0. [DOI] [PubMed] [Google Scholar]
  21. Gómez M. L., Ochatt C. M., Kazanietz M. G., Torres H. N., Téllez-Iñn M. T. Biochemical and immunological studies of protein kinase C from Trypanosoma cruzi. Int J Parasitol. 1999 Jul;29(7):981–989. doi: 10.1016/s0020-7519(99)00041-7. [DOI] [PubMed] [Google Scholar]
  22. Hidaka H., Kobayashi R. Pharmacology of protein kinase inhibitors. Annu Rev Pharmacol Toxicol. 1992;32:377–397. doi: 10.1146/annurev.pa.32.040192.002113. [DOI] [PubMed] [Google Scholar]
  23. Isola E. L., Lammel E. M., Giovanniello O., Katzin A. M., González Cappa S. M. Trypanosoma cruzi morphogenesis: preliminary purification of an active fraction from hemolymph and intestinal homogenate of Triatoma infestans. J Parasitol. 1986 Jun;72(3):467–469. [PubMed] [Google Scholar]
  24. Isola E. L., Lammel E. M., González Cappa S. M. Influencia de la hemolinfa de Triatoma infestans en la morfogénesis del Trypanosoma cruzi. Rev Argent Microbiol. 1983;15(3):181–185. [PubMed] [Google Scholar]
  25. Isola E. L., Lammel E. M., González Cappa S. M. Trypanosoma cruzi: differentiation after interaction of epimastigotes and Triatoma infestans intestinal homogenate. Exp Parasitol. 1986 Dec;62(3):329–335. doi: 10.1016/0014-4894(86)90039-1. [DOI] [PubMed] [Google Scholar]
  26. Isola E. L., Lammel E. M., Redruello M. Trypanosoma cruzi: conditions required to improve metacyclic differentiation in axenic culture. Rev Argent Microbiol. 1989 Jan-Mar;21(1):9–14. [PubMed] [Google Scholar]
  27. Kawamoto S., Hidaka H. 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets. Biochem Biophys Res Commun. 1984 Nov 30;125(1):258–264. doi: 10.1016/s0006-291x(84)80362-9. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Lammel E. M., Turyn D., Isola E. L. Trypanosoma cruzi: incorporation of macromolecule precursors during the morphogenic process. Rev Argent Microbiol. 1992 Apr-Jun;24(2):81–85. [PubMed] [Google Scholar]
  31. Leguizamón M. S., Campetella O. E., González Cappa S. M., Frasch A. C. Mice infected with Trypanosoma cruzi produce antibodies against the enzymatic domain of trans-sialidase that inhibit its activity. Infect Immun. 1994 Aug;62(8):3441–3446. doi: 10.1128/iai.62.8.3441-3446.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Newton A. C. Regulation of protein kinase C. Curr Opin Cell Biol. 1997 Apr;9(2):161–167. doi: 10.1016/s0955-0674(97)80058-0. [DOI] [PubMed] [Google Scholar]
  33. Nishizuka Y. Protein kinase C and lipid signaling for sustained cellular responses. FASEB J. 1995 Apr;9(7):484–496. [PubMed] [Google Scholar]
  34. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  35. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  37. Rodriguez-Pena A., Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. doi: 10.1016/s0006-291x(84)80213-2. [DOI] [PubMed] [Google Scholar]
  38. Takuwa N., Takuwa Y., Bollag W. E., Rasmussen H. The effects of bombesin on polyphosphoinositide and calcium metabolism in Swiss 3T3 cells. J Biol Chem. 1987 Jan 5;262(1):182–188. [PubMed] [Google Scholar]
  39. Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
  40. Way K. J., Chou E., King G. L. Identification of PKC-isoform-specific biological actions using pharmacological approaches. Trends Pharmacol Sci. 2000 May;21(5):181–187. doi: 10.1016/s0165-6147(00)01468-1. [DOI] [PubMed] [Google Scholar]
  41. Yunker C. E., Vaughn J. L., Cory J. Adaptation of an insect cell line (Grace's Antheraea cells) to medium free of insect hemolymph. Science. 1967 Mar 24;155(3769):1565–1566. doi: 10.1126/science.155.3769.1565. [DOI] [PubMed] [Google Scholar]
  42. de Isola E. L., Lammel E. M., Katzin V. J., Gonzalez Cappa S. M. Influence of organ extracts of Triatoma infestans on differentiation of Trypanosoma cruzi. J Parasitol. 1981 Feb;67(1):53–58. [PubMed] [Google Scholar]
  43. de Lema M. G., Aeberhard E. E. Desaturation of fatty acids in Trypanosoma cruzi. Lipids. 1986 Nov;21(11):718–720. doi: 10.1007/BF02537247. [DOI] [PubMed] [Google Scholar]

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

RESOURCES