Analytical and Bioanalytical Chemistry

, Volume 405, Issue 4, pp 1159–1170 | Cite as

MALDI imaging in human skin tissue sections: focus on various matrices and enzymes

  • Bernd Enthaler
  • Maria Trusch
  • Markus Fischer
  • Claudius Rapp
  • Julia K. Pruns
  • Jens-Peter Vietzke
Original Paper

Abstract

Matrix-assisted laser/desorption ionization (MALDI) mass-spectrometric imaging (MSI), also known as MALDI imaging, is a powerful technique for mapping biological molecules such as endogenous proteins and peptides in human skin tissue sections. A few groups have endeavored to apply MALDI-MSI to the field of skin research; however, a comprehensive article dealing with skin tissue sections and the application of various matrices and enzymes is not available. Our aim is to present a multiplex method, based on MALDI-MSI, to obtain the maximum information from skin tissue sections. Various matrices were applied to skin tissue sections: (1) 9-aminoacridine for imaging metabolites in negative ion mode; (2) sinapinic acid to obtain protein distributions; (3) α-cyano-4-hydroxycinnamic acid subsequent to on-tissue enzymatic digestion by trypsin, elastase, and pepsin, respectively, to localize the resulting peptides. Notably, substantial amounts of data were generated from the distributions retrieved for all matrices applied. Several primary metabolites, e.g. ATP, were localized and subsequently identified by on-tissue postsource decay measurements. Furthermore, maps of proteins and peptides derived from on-tissue digests were generated. Identification of peptides was achieved by elution with different solvents, mixing with α-cyano-4-hydroxycinnamic acid, and subsequent tandem mass spectrometry (MS/MS) measurements, thereby avoiding on-tissue MS/MS measurements. Highly abundant peptides were identified, allowing their use as internal calibrants in future MALDI-MSI analyses of human skin tissue sections. Elastin as an endogenous skin protein was identified only by use of elastase, showing the high potential of alternative enzymes. The results show the versatility of MALDI-MSI in the field of skin research. This article containing a methodological perspective depicts the basics for a comprehensive comparison of various skin states.

Figure

Matrix-assisted laser/desorption ionization (MALDI) mass-spectrometric imaging (MSI), also known as MALDI imaging, is a powerful technique for mapping biological molecules in human skin tissue sections. In this body of work, a multiplex method, based on MALDI-MSI, is presented to obtain maximum information from skin tissue sections. Therefore, various matrices were applied to skin tissue sections: (1) 9-aminoacridine (9-AA) for imaging small molecules in negative ion mode; (2) sinapinic acid (SA) to obtain protein distributions; (3) α-cyano-4-hydroxycinnamic acid (α-HCHA) subsequent to on-tissue enzymatic digestion by trypsin, elastase, and pepsin, respectively, to localize the resulting peptides. Of note, identification of metabolites was achieved by post-source decay (PSD) MALDI, and proteins were identified subsequent to enzymatic digestion via the resulting peptides which were eluted from the skin tissue section and afterwards analyzed with use of a tandem time-of-flight (ToF) mass spectrometer. The application of alternative enzymes, such as pepsin and elastase, is highlighted within this article

Keywords

Matrix-assisted laser/desorption ionization mass-spectrometric imaging Matrix-assisted laser/desorption ionization imaging Matrix-assisted laser/desorption ionization matrices Enzymes Skin tissue 

Notes

Acknowledgment

The authors thank Zorica Jovanovic for carefully reading of the manuscript.

Supplementary material

216_2012_6508_MOESM1_ESM.pdf (479 kb)
ESM 1(PDF 479 kb)

References

  1. 1.
    Karas M, Bachmann D, Bahr U, Hillenkamp F (1987) Int J Mass Spectrom Ion Process 78:53–68CrossRefGoogle Scholar
  2. 2.
    Karas M, Hillenkamp F (1988) Anal Chem 60:2299–2301CrossRefGoogle Scholar
  3. 3.
    Caprioli RM, Farmer TB, Gile J (1997) Anal Chem 69:4751–4760CrossRefGoogle Scholar
  4. 4.
    Gustafsson JO, Oehler MK, Ruszkiewicz A, McColl SR, Hoffmann P (2011) Int J Mol Sci 12:773–794CrossRefGoogle Scholar
  5. 5.
    Hanrieder J, Ljungdahl A, Andersson M (2012) J Vis Exp. doi:10.3791/3445
  6. 6.
    Wehder L, Ernst G, Crecelius AC, Guntinas-Lichius O, Melle C, Schubert US, von Eggeling F (2010) J Histochem Cytochem 58:929–937CrossRefGoogle Scholar
  7. 7.
    Hankin JA, Farias SE, Barkley RM, Heidenreich K, Frey LC, Hamazaki K, Kim HY, Murphy RC (2011) J Am Soc Mass Spectrom 22:1014–1021CrossRefGoogle Scholar
  8. 8.
    Hart PJ, Francese S, Claude E, Woodroofe MN, Clench MR (2011) Anal Bioanal Chem 401:115–125CrossRefGoogle Scholar
  9. 9.
    Berry KA, Hankin JA, Barkley RM, Spraggins JM, Caprioli RM, Murphy RC (2011) Chem Rev 111:6491–6512CrossRefGoogle Scholar
  10. 10.
    Goto-Inoue N, Hayasaka T, Zaima N, Setou M (2011) Biochim Biophys Acta 1811:961–969CrossRefGoogle Scholar
  11. 11.
    Castellino S, Groseclose MR, Wagner D (2011) Bioanalysis 3:2427–2441CrossRefGoogle Scholar
  12. 12.
    Cornett DS, Frappier SL, Caprioli RM (2008) Anal Chem 80:5648–5653CrossRefGoogle Scholar
  13. 13.
    Drexler DM, Tannehill-Gregg SH, Wang L, Brock BJ (2011) J Pharmacol Toxicol Methods 63:205–208CrossRefGoogle Scholar
  14. 14.
    Fehniger TE, Vegvari A, Rezeli M, Prikk K, Ross P, Dahlback M, Edula G, Sepper R, Marko-Varga G (2011) Anal Chem 83:8329–8336CrossRefGoogle Scholar
  15. 15.
    Avery JL, McEwen A, Flinders B, Francese S, Clench MR (2011) Xenobiotica 41:735–742CrossRefGoogle Scholar
  16. 16.
    Sugiura Y, Setou M (2010) J NeuroImmune Pharm 5:31–43CrossRefGoogle Scholar
  17. 17.
    Stoeckli M, Staab D, Schweitzer A, Gardiner J, Seebach D (2007) J Am Soc Mass Spectrom 18:1921–1924CrossRefGoogle Scholar
  18. 18.
    Lemaire R, Menguellet SA, Stauber J, Marchaudon V, Lucot JP, Collinet P, Farine MO, Vinatier D, Day R, Ducoroy P, Salzet M, Fournier I (2007) J Proteome Res 6:4127–4134CrossRefGoogle Scholar
  19. 19.
    Stauber J, Lemaire R, Franck J, Bonnel D, Croix D, Day R, Wisztorski M, Fournier I, Salzet M (2008) J Proteome Res 7:969–978CrossRefGoogle Scholar
  20. 20.
    Cazares LH, Troyer DA, Wang B, Drake RR, Semmes OJ (2011) Anal Bioanal Chem 401:17–27CrossRefGoogle Scholar
  21. 21.
    Schwamborn K, Krieg RC, Jirak P, Ott G, Knuchel R, Rosenwald A, Wellmann A (2010) J Cancer Res Clin Oncol 136:1651–1655CrossRefGoogle Scholar
  22. 22.
    Meding S, Nitsche U, Balluff B, Elsner M, Rauser S, Schone C, Nipp M, Maak M, Feith M, Ebert MP, Friess H, Langer R, Hofler H, Zitzelsberger H, Rosenberg R, Walch A (2012) J Proteome Res 11:1996–2003CrossRefGoogle Scholar
  23. 23.
    Taverna D, Nanney LB, Pollins AC, Sindona G, Caprioli R (2011) Exp Dermatol 20:642–647CrossRefGoogle Scholar
  24. 24.
    Enthaler B, Pruns JK, Wessel S, Rapp C, Fischer M, Wittern KP (2012) Anal Bioanal Chem 402:1159–1167CrossRefGoogle Scholar
  25. 25.
    Deininger SO, Cornett DS, Paape R, Becker M, Pineau C, Rauser S, Walch A, Wolski E (2011) Anal Bioanal Chem 401:167–181CrossRefGoogle Scholar
  26. 26.
    Cerruti CD, Benabdellah F, Laprevote O, Touboul D, Brunelle A (2012) Anal Chem 84:2164–2171CrossRefGoogle Scholar
  27. 27.
    Rieg S, Seeber S, Steffen H, Humeny A, Kalbacher H, Stevanovic S, Kimura A, Garbe C, Schittek B (2006) J Invest Dermatol 126:354–365CrossRefGoogle Scholar
  28. 28.
    Scott PG, Dodd CM, Tredget EE, Ghahary A, Rahemtulla F (1995) Histopathology 26:423–431CrossRefGoogle Scholar
  29. 29.
    Junqueira LC, Montes GS, Martins JE, Joazeiro PP (1983) Histochemistry 79:397–403CrossRefGoogle Scholar
  30. 30.
    Reed CC, Iozzo RV (2002) Glycoconj J 19:249–255CrossRefGoogle Scholar
  31. 31.
    Wright PA, Wilmouth RC, Clifton IJ, Schofield CJ (2001) Eur J Biochem 268:2969–2974CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Bernd Enthaler
    • 1
    • 2
  • Maria Trusch
    • 3
  • Markus Fischer
    • 2
  • Claudius Rapp
    • 1
  • Julia K. Pruns
    • 1
  • Jens-Peter Vietzke
    • 1
  1. 1.Beiersdorf AGHamburgGermany
  2. 2.Hamburg School of Food Science, Institute of Food ChemistryUniversity of HamburgHamburgGermany
  3. 3.Mass Spectrometry Facility, Institute of Organic ChemistryUniversity of HamburgHamburgGermany

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