Skip to main content
Log in

CD36 antisense expression in 3T3-F442A preadipocytes

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

Abstract

An adipocyte membrane glycoprotein, FAT, homologous to CD36, has been implicated in the binding/transport of long-chain fatty acids. FAT/CD36 was identified by reaction with reactive long chain fatty acids derivatives under conditions where they inhibited FA uptake. Expression of CD36 in fibroblasts lacking the protein led to induction of a saturable high affinity, phloretinsensitive component of oleate uptake. In this report, we have examined the effects of FAT/CD36 antisense expression in 3T3-F442A preadipocyte cells, on FA uptake and cell differentiation. Cells were transfected with pSG5-TAF vector obtained by insertion of antisense coding sequence of FAT/CD36 into the BamH 1 site of pSG5. Four clones were selected based on expression of antisense CD36 mRNA. Levels of CD36 protein were determined by flow cytometry and correlated with rates of oleate uptake. Three clones, TAF13, TAF25, and TAF38 exhibited low CD36 expression and one clone TAF 18 had expression comparable to that of F442A control cells. FA uptake rates in clones TAF13, TAF25 and TAF3 8 were lower than those observed in TAF18. At confluence, adipocyte differentiation could be promoted by addition of insulin and triiodothyronine only in TAF18 cells but not in TAF13, TAF25 or TAF38. Addition of fatty acids to clones TAF13, TAF25 and TAF38 lead to an induction of CD36 expression, an enhancement of FA uptake and better cell differentiation. The data support a role of CD36 in the membrane uptake of long chain FA. CD36 expression and FA uptake appear to be closely linked to preadipocyte differentiation.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Aburnrad NA, Perkins RC, Park M, Park CR: Mechanism of long chain fatty acid permeation in the isolated adipocyte. J Biol Chem 256: 9183–9191, 1981

    Google Scholar 

  2. Abumrad NA, Park JH, Park CR: Permeation of long chain fatty acids into adipocytes. J Biol Chem 259: 8945–8953, 1984

    Google Scholar 

  3. Schwieterman W, Sorrentino D, Potter BJ, Rand J, Kiang CL, Stump D, Berk PD: Uptake oleate by isolated rat adipocytes is mediated by a 40–kDa plasma membrane fatty acid binding protein closely related to that in liver and gut. Proc Natl Acad Sci USA 85: 359–363, 1988

    Google Scholar 

  4. Harmon CM, Luce P, Beth A, Abumrad NA: Labeling of adipocyte membranes by, Sulfo-N-Succinixndyl derivatives of long chain fatty acids: Inhibition of fatty acid transport. J Mem Biol 124: 261–268, 1991

    Google Scholar 

  5. Harmon CM, Aburnrad NA: Binding of sulfosuccinimidyl fatty acids to adipocyte membrane proteins: Isolation and amino-terminal sequence of an 88 kDa protein implicated in transport of fatty acids. J Mem Biol 131: 431–449, 1993

    Google Scholar 

  6. Oquendo P, Hundt E, Lawler J, Seed B: CD36 directly mediates cytoadherence of Plasmodium falcipamm parasitized erythrocytes. Cell 58(l): 95–101, 1989

    Google Scholar 

  7. Greenwalt DE, Lipsky RH, Ockenhouse CF, Ikeda H, Tandon NN, Jamieson GA: Membrane glycoprotein CD36: A review of its role in adherence, signal transduction, and transfusion medicine. Blood 80: 1105–1115, 1992

    Google Scholar 

  8. Aburnrad NA, El Maghrabi R, Amri EZ, Lopez E, Grimaldi P: Cloning of a rat adipocyte membrane protein implicated in binding or transport of long-chain fatty acids that is induced during adipocyte differentiation. Homology with human CD36. J Biol Chem 268: 17665–17668, 1993

    Google Scholar 

  9. Baillic AGS, Cobum CT, Aburnrad NA: Reversible binding of long-chain fatty acids to purified FAT, the adipose CD356 homolog. J Mem Biol 153: 75–81, 1996

    Google Scholar 

  10. Van Nieuwenhoven FA, Verstijnen CPW, Aburnrad NA, Willemsen PHM, Van Eys GJJM, Van Der Vusse GJ, Glatz MC: Putative membrane fatty acid translocase and cytoplasmic fatty acid binding protein are co-expressed in rat heail and skeletal muscles. Biochem Biophys Res Commun 207: 747–752, 1995

    Google Scholar 

  11. Sfeir Z, Ibrahimi A, Amri E, Grimaldi P, Abumrad NA: Regulation of FAT/CD36 gene expression: Further evidence in support of a role of the protein in fatty acid binding/transport. Prostaglandins, Leukotrienes and essential Fatty Acids 57: 17–21, 1997

    Google Scholar 

  12. Ibrahimi A, Sfeir Z, Magharaic H, Amri EZ, Grimaldi P, Abumrad NA: Expression of the CD36 homolog (FAT) in fibroblast cells: Effects on fatty acid transport. Proc Natl Acad Sci USA 93: 2646–2651, 1996

    Google Scholar 

  13. Chomezynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162(1): 156–159, 1987

    Google Scholar 

  14. Kashiwagi H, Tomiyama Y, Kosugi S, Shiraga M, Lipsky RH, Kanayarna Y, Kurata Y, Matsuzawa Y: Identification of molecular defects in a subject with type I CD36 deficiency. Blood 83(12): 3545–3552, 1994

    Google Scholar 

  15. Kashiwagi H, Tomlyama Y, Nozaki S, Honda S, Kosugi S, Shiraga M, Nak-agawa T, Nagao N, Kanakura Y, Kurata Y, Matsuzawa Y: A single nucleotide insertion in codon 317 of the CD36 gene leads to CD36 deficiency. Arterioscler Throm Vasc Biol 16(8): 1026–1032, 1996

    Google Scholar 

  16. Kashiwagi H, Tomiyama Y, Honda S, Kosuel S, Shiraga M, Nagao N, Sekiguchi S, Kanayarna Y, Kurata Y, Matsuzawa Y: Molecular basis of CD36 deficiency. Evidence that a 47SC—>T substitution (proline90—>serine) in CD36 cDNA accounts for CD36 deficiency. J Clin Inv (3): 1040–1046, 1995

  17. Tanaka T, Sohmiya K, Kawamura K: Is CD36 deficiency an etiology of hereditary hypertrophic cardiomyopathy? J Mol Cell Cardiol 29: 121–127, 1997

    Google Scholar 

  18. Tanaka T, Okamoto F, Sohmiya K, Kawamura K: Lack of myocardial iodine-123 15–(p-iodiphenyl)-3–R,S-methylpentadecanoic acid (BMIPP) uptake and CD36 abnormality—CD36 deficiency and hypertrophic cardiomyopathy. Japanese Circ J 61(8): 724–725, 1997

    Google Scholar 

  19. Inoue F, Hashimoto T, Nishida Y, Dohi K, Matsushima A, Sakakibara H, Ishida Y: Absence of myocardial 123I-BMIPP uptake in the presence of a normal coronary angiogram and normokinetics on a left ventriculogram. Japanese Circ J 61(3): 263–267, 1997

    Google Scholar 

  20. Endemann G, Stanton LW, Madden KS, Bryant CM, White RT, Protter AA: CD36 is receptor for oxidized low density lipoprotein. J Biol Chem 268: 11811–11816, 1993

    Google Scholar 

  21. Nozaki S, Kashiwagi H, Yamashita S, Nakagawa T, Kostner B, Tomiyama Y, Nakata A, Ishigarni M, Miyagawa J, Kameda-Takemura K, Kurata Y, Matsuzawa Y: Reduced uptake of oxidized low density lipoproteins in monocyte-derived macrophages from CD36–deficient subjects. J Clin Inv 96(4): 1859–1865, 1995

    Google Scholar 

  22. Stump DD, Zliou SL, Berk PD: Comparison of plasma membrane FABP and mitochondrial isoform of aspartate aminotransferase from rat liver. Am J Physiol 265(5 Pt 1): G894–G902, 1993

    Google Scholar 

  23. Shaffer JE, Lodish HF: Expression cloning and characterization of a novel adipocyte long chain fatty acid transport protein. Cell 79: 427–436, 1994

    Google Scholar 

  24. Berk PD, Zhou SL, Kiang CL, Stump D, Bradbury M, Isola LM: Uptake of long-chain fatty acid is selectively up-regulated in adipocytes of Zucker rats with genetic adiposity and noninsulin-dependent diabetes mellitus. J Biol Chem 2772: 8830–8835, 1997

    Google Scholar 

  25. Man MZ, Hui TY, Schaffer JE, Lodish HF, Bernlohr DA: Regulation of the murine adipocyte fatty acid transporter gene by insulin. Mol Endocrinol 10(8): 1021–1028, 1996

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sfeir, Z., Ibrahimi, A., Amri, Ez. et al. CD36 antisense expression in 3T3-F442A preadipocytes. Mol Cell Biochem 192, 3–8 (1999). https://doi.org/10.1023/A:1006811300125

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006811300125

Navigation