Skip to main content
SpringerLink
Log in

Single eye analysis and contralateral eye comparison of tear proteins in normal and dry eye model rabbits by MALDI-ToF mass spectrometry using wax-coated target plates

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A study of rabbit tear protein expression in a dry eye rabbit model was performed to determine if a pattern in expressed proteins could be identified. The uniqueness of the model allows the comparison of normal (control) eye tear protein expression with surgically induced dry eye tear protein expression in individual animals. The sensitivity of the method allows for single eye analysis. One-dimensional mini-gel electrophoresis of the tear proteins did not show substantial differences between band patterns of the normal versus the dry eye, but was used to assess the molecular weight ranges of the major proteins. Specific assignments of some of the predominant proteins were obtained by tandem mass spectrometry (MS) which showed that the lower molecular weight lipid-binding proteins (approximately 10 kDa to 36 kDa) constitute a considerable amount of the observed protein, followed in lesser quantities by the transferrins which have higher molecular weights ranging from 70 kDa to 85 kDa. Enhancement of matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) MS linear mode analysis of intact proteins in tear fluid was demonstrated through the use of wax-coated MALDI plates and spot washing. MALDI-ToF MS analysis of the expressed tear proteins illustrates that differences between normal eye tear and dry eye tear protein content are manifested in changes in the lower molecular weight lipid-binding proteins such as lipophilin which exhibits an increase in concentration in the dry eye, and β-2 microglobulin which undergoes a decrease.

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

  1. Holly FJ, Lemp MA (1973) Int Ophthalmol Clin 13:29

    Google Scholar 

  2. Ellison SA, Jacobson M, Levine MJ (1981) Lacrimal and salivary proteins. Immunol eye, workshop 3: immunological aspects of ocular disease: infection, inflammation, and allergy. Meeting Date 1980, 205

  3. Frey WH, DeSota-Johnson D, Hoffman C, McCall JT (1981) Am J Ophthalmol 92:559

    CAS  Google Scholar 

  4. Holly FJ, Hong BS (1982) Am J Optom Physiol Opt 59:43

    CAS  Google Scholar 

  5. Gachon AM, Richard J, Dastugue B (1982) Curr Eye Res 2:301

    Google Scholar 

  6. Suarez JC (1986) Ophthalmologica (Basel) 193:75

    CAS  Google Scholar 

  7. Kuizenga A, Stolwijk TR, van Agtmaal EJ, van Haeringen NJ, Kijlstra A (1990) Curr Eye Res 9:997

    CAS  Google Scholar 

  8. Gachon AMF (1994) Human lacrimal gland secretes proteins belonging to the group of hydrophobic molecule transporters. In: Sullivan DA (ed) Lacrimal gland, tear film, and dry eye syndromes. Plenum, New York, p 205

    Google Scholar 

  9. Baguet J, Claudon-Eyl V, Sommer F, Chevallier P (1995) CLAO J 21:114

    CAS  Google Scholar 

  10. Asrani AC, Lumsden AJ, Kumar R, Laurie GW (1998) Gene cloning of BM180, a lacrimal gland enriched basement membrane protein with a role in stimulated secretion. In: Sullivan DA, Dartt DA, Meneray MA (eds) Lacrimal gland, tear film, and dry eye syndromes 2. Plenum, New York, p 49

    Google Scholar 

  11. Lehrer RI, Xu G, Abduragimov A, Dinh NN, Qu XD, Martin D, Glasgow BJ (1998) FEBS Lett 432:163

    Article  CAS  Google Scholar 

  12. Schoenwald RD, Vidvauns S, Wurster DE, Barfknecht CF (1998) The role of tear proteins in tear film stability in the dry eye patient and in the rabbit. In: Sullivan DA, Dartt DA, Meneray MA (eds) Lacrimal gland, tear film, and dry eye syndromes 2. Plenum, New York, p 391

    Google Scholar 

  13. Boptom VE, Willcox MDP, Millar TJ (2000) Clin Exp Ophthalmol 28:208

    Article  Google Scholar 

  14. Moore JC, Tiffany JM (1981) Exp Eye Res 33:203

    Article  Google Scholar 

  15. Carrington SD, Hicks SJ, Corfield AP, Kaswan RL, Packer N, Bolis S, Morris CA (1998) Structural analysis of secreted ocular mucins in canine dry eye. In: Sullivan DA, Dartt DA, Meneray MA (eds) Lacrimal gland, tear film, and dry eye syndromes 2. Plenum, New York, p 253

    Google Scholar 

  16. McCulley JP, Shine WE (2001) Biosci Rep 21:407

    Article  CAS  Google Scholar 

  17. Salvatore MF, Pedroza L, Beuerman RW (1999) Curr Eye Res 18:455

    Article  CAS  Google Scholar 

  18. Barrera R, Jimenez A, Lopez R, Mane MC, Rodriguez JF, Molleda JM (1992) Am J Vet Res 53:454

    CAS  Google Scholar 

  19. Kondo Y (1994) Exp Anim 43:1

    CAS  Google Scholar 

  20. Berta A (1982) Graefe Arch Clin Exp Ophthalmol 219:95

    Article  CAS  Google Scholar 

  21. Joo W, Lee D, Kim C (2003) Biosci Biotechnol Biochem 67:1574

    Article  CAS  Google Scholar 

  22. Varnell RJ, Maitchouk DY, Beuerman RW, Salvatore MF, Carlton JE, Haag AM (1997) J Cap Elec 1:1

    Google Scholar 

  23. Boonstra A, Kijlstra A (1984) Curr Eye Res 3:1461

    CAS  Google Scholar 

  24. Wollensak G, Mur E, Mayr A, Baier G, Gottinger W, Stoffler G (1990) Graefe Arch Clin Exp Ophthalmol 228:78–82

    CAS  Google Scholar 

  25. Gachon AM, Verrelle P, Betail G, Dastugue B (1979) Exp Eye Res 29:539

    Article  CAS  Google Scholar 

  26. Boonstra A, Kijlstra A (1984) Exp Eye Res 38:561

    Article  CAS  Google Scholar 

  27. Inada K, Baba H, Okamura R (1982) Jpn J Ophthalmol 26:314

    CAS  Google Scholar 

  28. Schmut O, Horwath-Winter J, Zenker A, Trummer G (2002) Graefe Arch Clin Exp Ophthalmol 240:900

    CAS  Google Scholar 

  29. Glasgow BJ, Abduragimov AR, Farahbakhsh ZT, Faull KF, Hubbell WL (1995) Curr Eye Res 14:363

    CAS  Google Scholar 

  30. Zhou L, Beuerman RW, Barathi A, Tan D (2003) Rapid Commun Mass Spectrum 17:401

    Article  CAS  Google Scholar 

  31. Mulvenna I, Stapleton F, Hains PG, Cengiz A, Tan M, Walsh B, Holden B (2000) Clin Exp Ophthalmol 28:205

    Article  CAS  Google Scholar 

  32. Fung K, Morris C, Duncan M (2002) Mass spectrometric techniques applied to the analysis of human tears: a focus on the peptide and protein constituents. In: Sullivan DA, Calmenson SA, Stern ME, Dartt DA (eds) Lacrimal gland, tear film, and dry eye syndromes 3. Kluwer Academic/Plenum, New York, p 601

    Google Scholar 

  33. Cao P, Moini M (1998) J Am Soc Mass Spectrom 9:1081

    Article  CAS  Google Scholar 

  34. Bonavida B, Sapse AT, Sercarz EE (1968) Invest Ophthalmol 7:435

    CAS  Google Scholar 

  35. Dartt LA, Botelho SY (1979) Invest Ophthalmol Vis Sci 18:1207

    CAS  Google Scholar 

  36. Dohlman CH, Friend J, Kalevar V, Yogoda D, Balazs E (1976) Exp Eye Res 22:359

    Article  CAS  Google Scholar 

  37. Ng V, Cho P, To C (2000) Graefe Arch Clin Exp Ophthalmol 238:738

    Article  CAS  Google Scholar 

  38. Lemp MA (1995) CLAO J 21:221

    CAS  Google Scholar 

  39. Pflugfelder SC, Solomon A, Stern ME (2000) Cornea 19:644

    Article  CAS  Google Scholar 

  40. Schaumberg DA, Buring JE, Sullivan DA, Dana MR (2001) JAMA 286:2114

    Article  CAS  Google Scholar 

  41. Ohashi Y, Ishida R, Kojima T, Goto E, Matsumoto Y, Watanabe K, Ishida N, Nakat K, Takeuchi T, Tsubota K (2003) Am J Ophthalmol 136:291

    Article  CAS  Google Scholar 

  42. Nguyen DH, Beuerman RW, Meneray MA, Maitchouk D (1998) Sensory denervation leads to deregulated protein synthesis in the lacrimal gland. In: Sullivan DA, Dartt DA, Meneray MA (eds) Lacrimal gland, tear film, and dry eye syndromes 2. Plenum, New York, p 55

    Google Scholar 

  43. Varnell RJ, Maitchouk DM, Beuerman RW, Carlton JE, Haag A (1998) Small-volume analysis of rabbit tears and effects of a corneal wound on tear protein spectra. In: Sullivan DA, Dartt DA, Meneray MA (eds) Lacrimal gland, tear film, and dry eye syndromes 2. Plenum, New York, p 659

    Google Scholar 

  44. Folch J, Lees M, Stanly GHS (1957) J Biol Chem 226:497

    CAS  Google Scholar 

  45. Bligh EG, Dyer WJ (1959) Can J Biochem Physiol 37:911

    CAS  Google Scholar 

  46. Shevchenko A, Wilm M, Vorm O, Mann M (1996) Anal Chem 68:850–858

    Article  CAS  Google Scholar 

  47. Smirnov IP, Zhu X, Taylor T, Huang Y, Ross P, Papayanopoulos IA, Martin SA, Pappin DJ (2004) Anal Chem 76:2958

    Article  CAS  Google Scholar 

  48. Terry DE, Umstot E, Desiderio DM (2004) J Am Soc Mass Spectrom 15:784

    Article  CAS  Google Scholar 

  49. Lehrer RI, Xu G, Abduragimov A, Dinh NN, Qu XD, Martin D, Glasgow BJ (1998) FEBS Lett 432:163

    Article  CAS  Google Scholar 

  50. Lehrer RI, Nguyen T, Zhao C, Ha CX, Glasgow BJ (2000) Ann N Y Acad Sci 923:59

    Article  CAS  Google Scholar 

  51. Ham BM, Jacob JT, Keese MM, Cole RB (2004) J Mass Spectrom 39:1321

    Article  CAS  Google Scholar 

  52. Holzfeind P, Merschak P, Dieplinger H, Bernhard R (1995) Exp Eye Res 61:495

    Article  CAS  Google Scholar 

  53. Azen EA (1976) Biochem Genet 14:225

    Article  CAS  Google Scholar 

  54. Azzarolo AM, Brew K, Kota S, Ponomareva O, Schwartz J, Zylberberg C (2004) Comp Biochem Physiol, B 138:111

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Financial support was provided by the Louisiana Board of Regents through grants HEF(2001-06)-08 and LEQSF(2001-04)-RD-B-11, and from the National Eye Institute through NEI/R03 EY014021, and an unrestricted grant from Research to Prevent Blindness, New York, NY.

Author information

Authors and Affiliations

  1. Department of Chemistry, University of New Orleans, 2000 Lakeshore Drive, New Orleans, LA, 70148, USA

    Bryan M. Ham & Richard B. Cole

  2. Department of Ophthalmology, Louisiana State University Health Sciences Center, 2020 Gravier Street, Suite B, New Orleans, LA, 70112, USA

    Jean T. Jacob

Authors
  1. Bryan M. Ham
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jean T. Jacob
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard B. Cole
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Richard B. Cole.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ham, B.M., Jacob, J.T. & Cole, R.B. Single eye analysis and contralateral eye comparison of tear proteins in normal and dry eye model rabbits by MALDI-ToF mass spectrometry using wax-coated target plates. Anal Bioanal Chem 387, 889–900 (2007). https://doi.org/10.1007/s00216-006-1018-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-006-1018-9

Keywords

Search

Navigation