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The discovery of stannin in rat dorsal root ganglia using an integrated proteohistological approach
Clinical Proteomics volume 1, pages249–258(2004)
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.
Merchant, M. and Weinberger, S.R. (2000). Recent advancements in surface-enhanced laser desorption/ionization-time of flight-mass spectrometry. Electrophoresis 21:1164–1177.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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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) doi:10.1385/CP:1:3-4:249
- Ciphergen ProteinChip®
- dorsal root ganglion
- in situ hybridization