Skip to main content
  • Original Article
  • Open access
  • Published:

The discovery of stannin in rat dorsal root ganglia using an integrated proteohistological approach

Potential role in inflammatory hyperalgesia

Abstract

The use of proteomic analysis to discover proteins (previously identified or unknown) in a tissue sample is a valuable tool. However, there is a limit to the extent one can validate a discovery with any single technology. In an effort to obviate this inherent constraint and to add value and dimension to protein profiling, we have coupled the information obtained through proteomic techniques with the validation provided by in situ hybridization and immunohistochemistry techniques. This approach can be illustrated by our efforts in the discovery of stannin in rat dorsal root ganglia (DRG). In this study, we initially used the Ciphergen ProteinChip® to perform protein profiling on the DRG of rats in a carrageenan-induced paw inflammation study. In an effort to discover new potential targets in inflammatory pain models, we profiled many potential peaks unique to the ipsilateral DRG of interest. One protein, found to bind to a hydrophobic chip at a molecular mass of 9500 Dalton, was preliminarily identified as stannin. To confirm its identification, we performed in situ hybridization and immunohistochemistry on the source DRG tissue to investigate the presence of stannin mRNA and protein expression, respectively. In addition to confirming the presence of stannin in these DRGs, we observed the upregulation of stannin in the DRGs over the course of carrageenan-induced inflammation, suggesting a possible role of stannin in inflammatory hyperalgesia. Taken together, these results illustrate the synergistic benefits of coupling 0 proteomic and histochemical techniques in identifying and validating targets and biomarkers for drug discovery.

References

  1. Merchant, M. and Weinberger, S.R. (2000). Recent advancements in surface-enhanced laser desorption/ionization-time of flight-mass spectrometry. Electrophoresis 21:1164–1177.

    Article  PubMed  CAS  Google Scholar 

  2. Rohde, E., Tomlinson, A.J., Johnson, D.H., and Naylor, S. (1998). Comparison of protein mixtures in aqueous humor by membrane preconcentration—capillary electrophoresis—mass spectrometry. Electrophoresis 19:2361–2370.

    Article  PubMed  CAS  Google Scholar 

  3. Nantel F., Denis, D., Gordon, R., Northey, A., Cirino, M., Metters, K.M., et al. (1999). Distribution and regulation of cyclooxygenase-2 in carrageenan-induced inflammation. Br. J. Pharmacol. 128:853–859.

    Article  PubMed  CAS  Google Scholar 

  4. Black, J.A., Liu, S., Tanaka, M., Cummins, T.R., and Waxman, S.G. (2004) Changes in the expression of tetrodotoxin-sensitive sodium channels within dorsal root ganglia neurons in inflammatory pain. Pain 108:237–247.

    Article  PubMed  CAS  Google Scholar 

  5. Ji, R.R., Zhang, Q., Law, P.Y., Low, H.H., Elde, R., and Hokfelt, T. (1995). Expression of mu-, delta-, and kappa-opioid receptor-like immunoreactivities in rat dorsal root ganglia after carrageenan-induced inflammation. J. Neurosci. 15:8156–8166.

    PubMed  CAS  Google Scholar 

  6. Sweitzer, S.M., Colburn, R.W., Rutkowski, M., and DeLeo, J.A. (1999). Acute peripheral inflammation induces moderate glial activation and spinal IL-1beta expression that correlates with pain behavior in the rat. Brain Res. 829:209–221.

    Article  PubMed  CAS  Google Scholar 

  7. Toggas, S.M., Krady, J.K., and Billingsley, M.L. (1992). Molecular neurotoxicology of trimethyltin: identification of stannin, a novel protein expressed in trimethyltin-sensitive cells. Mol. Pharmacol. 42:44–56.

    PubMed  CAS  Google Scholar 

  8. Toggas, S.M., Krady, J.K., Thompson, T.A., and Billingsley, M.L. (1993). Molecular mechanisms of selective neurotoxicants: studies on organotin compounds. Ann. NY Acad. Sci. 679:157–177.

    Article  PubMed  CAS  Google Scholar 

  9. Thompson, T.A., Lewis, J.M., Dejneka, N.S., Severs, W.B., Polavarapu, R., and Billingsley, M.L. (1996). Induction of apoptosis by organotin compounds in vitro: neuronal protection with antisense oligonucleotides directed against stannin. J. Pharmacol. Exp. Ther. 276:1201–1216.

    PubMed  CAS  Google Scholar 

  10. Hargreaves, K., Dubner, R., Brown, F., Flores, C., and Joris, J. (1988). A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain 32:77–88.

    Article  PubMed  CAS  Google Scholar 

  11. Dirig, D.M., Salami, A., Rathbun, M.L., Ozaki, G.T., and Yaksh, T.L. (1997). Characterization of variables defining hindpaw withdrawal latency evoked by radiant thermal stimuli. J. Neurosci. Methods 76:183–191.

    Article  PubMed  CAS  Google Scholar 

  12. Wilkins, M.R., Gasteiger, E., Bairoch, A., Sanchez, J.-C., Williams, K.L., Appel, R.D., et al. (1999). Protein identification and analysis tools in the ExPASy server, in Methods in Molecular Biology, vol; 112 (Link, A.J., ed.), Humana Press, Totowa, NJ, pp. 531–552.

    Google Scholar 

  13. Dejneka, N.S., Patanow, C.M., Polavarapu, R., Toggas, S.M., Krady, J.K., and Billingsley, M.L. (1997). Localization and characterization of stannin: relationship to cellular sensitivity to organotin compounds. Neurochem. Int. 31: 801–815.

    Article  PubMed  CAS  Google Scholar 

  14. Cunha, F.Q., Poole, S., Lorenzetti, B.B., and Ferreira, S.H. (1992). The pivotal role of tumour necrosis factor alpha in the development of inflammatory hyperalgesia. Br. J. Pharmacol. 107:660–664.

    PubMed  CAS  Google Scholar 

  15. Horrevoets, A.J., Fontijn, R.D., van Zonneveld, A.J., de Vries, C.J., ten Cate, J.W., and Pannekoek, H. (1999). Vascular endothelial genes that are responsive to tumor necrosis factor-alpha in vitro are expressed in atherosclerotic lesions, including inhibitor of apoptosis protein-1, stannin, and two novel genes. Blood 93:3418–3431.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Stanley M. Belkowski.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Belkowski, S.M., Colburn, R.W., Stone, D.J. et al. The discovery of stannin in rat dorsal root ganglia using an integrated proteohistological approach. Clin Proteom 1, 249–258 (2004). https://doi.org/10.1385/CP:1:3-4:249

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1385/CP:1:3-4:249

Key Words