Skip to main content
SpringerLink
Log in

Analysis of ABCC6 (MRP6) in normal human tissues

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

To determine the tissue distribution of the ABC transporter ABCC6 in normal human tissues, we analyzed tissue arrays for the presence of ABCC6 mRNA by in situ hybridization and ABCC6 protein by immunohistochemistry using the polyclonal antibody HB-6. We detected ABCC6 mRNA and protein in various epithelial cells of exocrine and endocrine tissues, such as acinar cells in the pancreas, mucosal cells of the intestine and follicular epithelial cells of the thyroid. We obtained a very strong immunostaining for enteroendocrine G cells in the stomach. In addition, ABCC6 mRNA and protein were present in most neurons of the brain, in alveolar macrophages in the lungs and lymphocytes in the lymph node. Immunohistochemisty using the monoclonal antibody M6II-31 confirmed the widespread tissue distribution of ABCC6. The physiological substrate(s) of ABCC6 are yet unknown, but we suggest that ABCC6 fulfills multiple functions in different tissues. The strong immunostaining for ABCC6 in G cells suggests that it plays an important role in these endocrine 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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Akabas MH (2000) Cystic fibrosis transmembrane conductance regulator. Structure and function of an epithelial chloride channel. J Biol Chem 275:3729–3732

    CAS  PubMed  Google Scholar 

  • Baschong W, Suetterlin R, Laeng RH (2001) Control of autofluorescence of archival formaldehyde-fixed, paraffin-embedded tissue in confocal laser scanning microscopy (CLSM). J Histochem Cytochem 49:1565–1572

    CAS  PubMed  Google Scholar 

  • Beck K, Hayashi K, Nishiguchi B, Le Saux O, Hayashi M, Boyd CD (2003) The distribution of Abcc6 in normal mouse tissues suggests multiple functions for this ABC transporter. J Histochem Cytochem 51:887–902

    CAS  PubMed  Google Scholar 

  • Belinsky MG, Chen ZS, Shchaveleva I, Zeng H, Kruh GD (2002) Characterization of the drug resistance and transport properties of multidrug resistance protein 6 (MRP6, ABCC6). Cancer Res 62:6172–6177

    CAS  PubMed  Google Scholar 

  • Bergen AA, Plomp AS, Schuurman EJ, Terry S, Breuning M, Dauwerse H, Swart J, Kool M, van Soest S, Baas F, ten Brink JB, de Jong PT (2000) Mutations in ABCC6 cause pseudoxanthoma elasticum. Nat Genet 25:228–231

    CAS  PubMed  Google Scholar 

  • Bordi C, D’Adda T, Azzoni C, Ferraro G (2000) Classification of gastric endocrine cells at the light and electron microscopical levels. Microsc Res Tech 48:258–271

    Article  CAS  PubMed  Google Scholar 

  • Borst P, Evers R, Kool M, Wijnholds J (1999) The multidrug resistance protein family. Biochim Biophys Acta 1461:347–357

    CAS  PubMed  Google Scholar 

  • Darby M, Kuzmiski JB, Panenka W, Feighan D, MacVicar BA (2003) ATP released from astrocytes during swelling activates chloride channels. J Neurophysiol 89:1870–1877

    CAS  PubMed  Google Scholar 

  • De Cory HH, Piech-Dumas KM, Sheu SS, Federoff HJ, Anders MW (2001) Efflux of glutathione conjugate of monochlorobimane from striatal and cortical neurons. Drug Metab Dispos 29:1256–1262

    PubMed  Google Scholar 

  • Dockray GJ, Varro A, Dimaline R (1996) Gastric endocrine cells: gene expression, processing, and targeting of active products. Physiol Rev 76:767–798

    CAS  PubMed  Google Scholar 

  • Flens MJ, Zaman GJ, van der Valk P, Izquierdo MA, Schroeijers AB, Scheffer GL, van der Groep P, de Haas M, Meijer CJ, Scheper RJ (1996) Tissue distribution of the multidrug resistance protein. Am J Pathol 148:1237–1247

    CAS  PubMed  Google Scholar 

  • Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58

    CAS  PubMed  Google Scholar 

  • Hayashi K, Hayashi M, Jalkanen M, Firestone JH, Trelstad RL, Bernfield M (1987) Immunocytochemistry of cell surface heparan sulfate proteoglycan in mouse tissues. A light and electron microscopic study. J Histochem Cytochem 35:1079–1088

    CAS  PubMed  Google Scholar 

  • Hirrlinger J, Schulz JB, Dringen R (2002) Glutathione release from cultured brain cells: multidrug resistance protein 1 mediates the release of GSH from rat astroglial cells. J Neurosci Res 69:318–326

    Article  CAS  PubMed  Google Scholar 

  • Hu X, Plomp A, Wijnholds J, Ten Brink J, Van Soest S, Van Den Born LI, Leys A, Peek R, De Jong PT, Bergen AA (2003) ABCC6/MRP6 mutations: further insight into the molecular pathology of pseudoxanthoma elasticum. Eur J Hum Genet 11:215–224

    Article  CAS  PubMed  Google Scholar 

  • Ilias A, Urban Z, Seidl TL, Le Saux O, Sinko E, Boyd CD, Sarkadi B, Varadi A (2002) Loss of ATP-dependent transport activity in pseudoxanthoma elasticum- associated mutants of human ABCC6 (MRP6). J Biol Chem 277:16860–16867

    Article  CAS  PubMed  Google Scholar 

  • Jedlitschky G, Leier I, Buchholz U, Center M, Keppler D (1994) ATP-dependent transport of glutathione S-conjugates by the multidrug resistance-associated protein. Cancer Res 54:4833–4836

    CAS  PubMed  Google Scholar 

  • Ko YH, Pedersen PL (2001) Cystic fibrosis: a brief look at some highlights of a decade of research focused on elucidating and correcting the molecular basis of the disease. J Bioenerg Biomembr 33:513–521

    Article  CAS  PubMed  Google Scholar 

  • Komminoth P (1992) Digoxigenin as an alternative probe labeling for in situ hybridization. Diagn Mol Pathol 1:142–150

    CAS  PubMed  Google Scholar 

  • Kool M, van der Linden M, de Haas M, Baas F, Borst P (1999) Expression of human MRP6, a homologue of the multidrug resistance protein gene MRP1, in tissues and cancer cells. Cancer Res 59:175–182

    CAS  PubMed  Google Scholar 

  • Kruh GD, Zeng H, Rea PA, Liu G, Chen ZS, Lee K, Belinsky MG (2001) MRP subfamily transporters and resistance to anticancer agents. J Bioenerg Biomembr 33:493–501

    Article  CAS  PubMed  Google Scholar 

  • Larsson LI (1981) Adrenocorticotropin-like and alpha-melanotropin-like peptides in a subpopulation of human gastrin cell granules: bioassay, immunoassay, and immunocytochemical evidence. Proc Natl Acad Sci USA 78:2990–2994

    CAS  PubMed  Google Scholar 

  • Le Saux O, Beck K, Sachsinger C, Silvestri C, Treiber C, Goring HH, Johnson EW, De Paepe A, Pope FM, Pasquali-Ronchetti I, Bercovitch L, Marais AS, Viljoen DL, Terry SF, Boyd CD (2001) A spectrum of ABCC6 mutations is responsible for pseudoxanthoma elasticum. Am J Hum Genet 69:749–764

    Article  CAS  PubMed  Google Scholar 

  • Le Saux O, Urban Z, Tschuch C, Csiszar K, Bacchelli B, Quaglino D, Pasquali-Ronchetti I, Pope FM, Richards A, Terry S, Bercovitch L, de Paepe A, Boyd CD (2000) Mutations in a gene encoding an ABC transporter cause pseudoxanthoma elasticum. Nat Genet 25:223–227

    Article  CAS  PubMed  Google Scholar 

  • Lebwohl M, Schwartz E, Lemlich G, Lovelace O, Shaikh-Bahai F, Fleischmajer R (1993) Abnormalities of connective tissue components in lesional and non-lesional tissue of patients with pseudoxanthoma elasticum. Arch Dermatol Res 285:121–126

    Article  CAS  PubMed  Google Scholar 

  • Leier I, Jedlitschky G, Buchler M, Buchholz U, Brom M, Keppler D (1996) Identification of the biosynthetic leukotriene C4 export pump in murine mastocytoma cells as a homolog of the multidrug-resistance protein. Eur J Biochem 242:201–205

    CAS  PubMed  Google Scholar 

  • Loe DW, Almquist KC, Cole SP, Deeley RG (1996) ATP-dependent 17 beta-estradiol 17-(beta-D-glucuronide) transport by multidrug resistance protein (MRP). Inhibition by cholestatic steroids. J Biol Chem 271:9683–9689

    Article  CAS  PubMed  Google Scholar 

  • Madon J, Hagenbuch B, Landmann L, Meier PJ, Stieger B (2000) Transport function and hepatocellular localization of mrp6 in rat liver. Mol Pharmacol 57:634–641

    CAS  PubMed  Google Scholar 

  • Meloni I, Rubegni P, De Aloe G, Bruttini M, Pianigiani E, Cusano R, Seri M, Mondillo S, Federico A, Bardelli AM, Andreassi L, Fimiani M, Renieri A (2001) Pseudoxanthoma elasticum: point mutations in the ABCC6 gene and a large deletion including also ABCC1 and MYH11. Hum Mutat 18:85

    Article  CAS  PubMed  Google Scholar 

  • Mendelsohn G, Bulkley BH, Hutchins GM (1978) Cardiovascular manifestations of pseudoxanthoma elasticum. Arch Pathol Lab Med 102:298–302

    CAS  PubMed  Google Scholar 

  • Muller M, Meijer C, Zaman GJ, Borst P, Scheper RJ, Mulder NH, de Vries EG, Jansen PL (1994) Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport. Proc Natl Acad Sci USA 91:13033–13037

    CAS  PubMed  Google Scholar 

  • Neldner KH (1988) Pseudoxanthoma elasticum. Int J Dermatol 27:98–100

    CAS  PubMed  Google Scholar 

  • Nishida H, Endo M, Koyanagi H, Ichihara T, Takao A, Maruyama M (1990) Coronary artery bypass in a 15-year-old girl with pseudoxanthoma elasticum. Ann Thorac Surg 49:483–485

    CAS  PubMed  Google Scholar 

  • Pulaski L, Jedlitschky G, Leier I, Buchholz U, Keppler D (1996) Identification of the multidrug-resistance protein (MRP) as the glutathione-S-conjugate export pump of erythrocytes. Eur J Biochem 241:644–648

    Article  CAS  PubMed  Google Scholar 

  • Ringpfeil F, Lebwohl MG, Christiano AM, Uitto J (2000) Pseudoxanthoma elasticum: mutations in the MRP6 gene encoding a transmembrane ATP-binding cassette (ABC) transporter. Proc Natl Acad Sci USA 97:6001–6006

    Article  CAS  PubMed  Google Scholar 

  • Sachs G, Zeng N, Prinz C (1997) Physiology of isolated gastric endocrine cells. Annu Rev Physiol 59:243–256

    CAS  PubMed  Google Scholar 

  • Scheffer GL, Hu X, Pijnenborg AC, Wijnholds J, Bergen AA, Scheper RJ (2002) MRP6 (ABCC6) detection in normal human tissues and tumors. Lab Invest 82:515–518

    CAS  PubMed  Google Scholar 

  • Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333

    CAS  PubMed  Google Scholar 

  • Struk B, Cai L, Zach S, Ji W, Chung J, Lumsden A, Stumm M, Huber M, Schaen L, Kim CA, Goldsmith LA, Viljoen D, Figuera LE, Fuchs W, Munier F, Ramesar R, Hohl D, Richards R, Neldner KH, Lindpaintner K (2000) Mutations of the gene encoding the transmembrane transporter protein ABC-C6 cause pseudoxanthoma elasticum. J Mol Med 78:282–286

    Article  CAS  PubMed  Google Scholar 

  • Taipalensuu J, Tornblom H, Lindberg G, Einarsson C, Sjoqvist F, Melhus H, Garberg P, Sjostrom B, Lundgren B, Artursson P (2001) Correlation of gene expression of ten drug efflux proteins of the ATP- binding cassette transporter family in normal human jejunum and in human intestinal epithelial Caco-2 cell monolayers. J Pharmacol Exp Ther 299:164–170

    CAS  PubMed  Google Scholar 

  • Uitto J, Boyd CD, Lebwohl MG, Moshell AN, Rosenbloom J, Terry S (1998) International Centennial Meeting on Pseudoxanthoma Elasticum: progress in PXE research. J Invest Dermatol 110:840–842

    Article  CAS  PubMed  Google Scholar 

  • Uitto J, Shamban A (1987) Heritable skin diseases with molecular defects in collagen or elastin. Dermatol Clin 5:63–84

    CAS  PubMed  Google Scholar 

  • Weenink AC, Dijkman G, de Meijer PH (1996) Pseudoxanthoma elasticum and its complications: two case reports. Neth J Med 49:24–29

    Article  CAS  PubMed  Google Scholar 

  • Wijnholds J, Evers R, van Leusden MR, Mol CA, Zaman GJ, Mayer U, Beijnen JH, van der Valk M, Krimpenfort P, Borst P (1997) Increased sensitivity to anticancer drugs and decreased inflammatory response in mice lacking the multidrug resistance-associated protein. Nat Med 3:1275–1279

    CAS  PubMed  Google Scholar 

  • Zhang Y, Han H, Elmquist WF, Miller DW (2000) Expression of various multidrug resistance-associated protein (MRP) homologues in brain microvessel endothelial cells. Brain Res 876:148–153

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by NIH grants EY13019 and RR16453 to Charles D. Boyd. We thank Dr. Andras Varadi (Institute of Enzymology, Hungarian Academy of Sciences) for many valuable discussions regarding the possible functions of ABCC6.

Author information

Authors and Affiliations

  1. Laboratory of Matrix Pathobiology, The Pacific Biomedical Research Center, University of Hawai’i, 1993 East-West Road, Honolulu, HI, 96822, USA

    Konstanze Beck, Kimiko Hayashi, Ka’ohimanu Dang, Masando Hayashi & Charles D. Boyd

  2. Centre for Vascular Research, School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia

    Konstanze Beck

Authors
  1. Konstanze Beck
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kimiko Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ka’ohimanu Dang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Masando Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Charles D. Boyd
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Konstanze Beck.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Beck, K., Hayashi, K., Dang, K. et al. Analysis of ABCC6 (MRP6) in normal human tissues. Histochem Cell Biol 123, 517–528 (2005). https://doi.org/10.1007/s00418-004-0744-3

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-004-0744-3

Keywords

Search

Navigation