Phosphoproteomic Analysis of Neurotrophin Receptor TrkB Signaling Pathways in Mouse Brain
- 170 Downloads
1. The signaling pathways activated by trkB neurotrophin receptor have been studied in detail in cultured neurons, but little is known about the pathways activated by trkB in intact brain. TrkB is a tyrosine kinase and protein phosphorylation is a key regulatory process in the neuronal signal transduction pathways.
2. We have investigated trkB signaling in the transgenic mice overexpressing trkB in postnatal neurons (trkB.TK) using phosphoproteomics.
3. We found that several proteins are overphosphorylated on tyrosine residues in the brain of trkB.TK mice and identified some of these proteins.
4. We demonstrate that the well characterized signaling molecules mitogen-activated protein kinase (MAPK) and cyclic AMP responsive element binding protein (CREB) were phosphorylated at a higher level in the brain of trkB.TK mice when compared to the wild type littermates. Furthermore, we found that β-actin was tyrosine phosphorylated in the brain of the transgenic mice.
5. Our results demonstrate that phosphoproteomics is a sensitive approach to investigate signaling pathways activated in mouse brain.
KEY WORDS:BDNF phosphorylation MAPK CREB actin proteomics
Mass spectrometric protein identifications were performed at the Protein Chemistry Research Group and Core Facility, Institute of Biotechnology,University of Helsinki. We would like to thank MSc. Saara Ihalainen, Dr. Nisse Kalkkinen and Dr. Gunilla Rönnholm for their help in the analysis and Dr. Moshe Finel for his help with electrophoresis.
- Baba, T., Fusaki, N., Shinya, N., Iwamatsu, A., and Hozumi, N. (2003). Actin tyrosine dephosphorylation by the Src homology 1-containing protein tyrosine phosphatase is essential for actin depolymerization after membrane IgM cross-linking. J. Immunol. 170:3762–3768.Google Scholar
- Koponen, E., Voikar, V., Riekki, R., Saarelainen, T., Rauramaa, T., Rauvala, H., Taira, T., and Castrén, E. (2004b). Transgenic mice overexpressing the full-length neurotrophin receptor TrkB exhibit increased activation of the TrkB-PLCgamma pathway, reduced anxiety, and facilitated learning. Mol. Cell Neurosci. 26:166–181.CrossRefPubMedGoogle Scholar
- Lim, Y. P., Wong, C. Y., Ooi, L. L., Druker, B. J., and Epstein, R. J. (2004). Selective tyrosine hyperphosphorylation of cytoskeletal and stress proteins in primary human breast cancers: implications for adjuvant use of kinase-inhibitory drugs. Clin. Cancer Res. 10:3980–3987.CrossRefPubMedGoogle Scholar
- Saarelainen, T., Hendolin, P., Lucas, G., Koponen, E., Sairanen, M., MacDonald, E., Agerman, K., Haapasalo, A., Nawa, H., Aloyz, R., Ernfors, P., and Castrén, E. (2003). Activation of the TrkB neurotrophin receptor is induced by antidepressant drugs and is required for antidepressant-induced behavioral effects. J. Neurosci. 23:349–357.PubMedGoogle Scholar