Spleen tyrosine kinase (Syk) is expressed widely in hematopoietic and non-hematopoietic cells. The widespread distribution of Syk and its involvement in host defense and allergic reactions, prompted us analyze the influence of microbial exposure on Syk expression. We compared the distribution of Syk in various tissues of germ-free and conventional mice using immunohistochemistry, Western blot analysis and real time RT-PCR. Total Syk expression was similar between germ-free and conventional mice. Since it has been claimed that Syk isoforms are differentially expressed, we studied the distribution and abundance of Syk (L) and Syk (S) isoforms in tissues from these mice. In contrast to previous reports, we found broad tissue expression of Syk (S). Interestingly, in germ-free mice the amount of Syk (S) but not Syk L protein was selectively increased in lung and spleen. In summary, our study reveals new and broad tissue expression of both Syk isoforms and demonstrates that lack of microbial flora results in selectively increased expression of Syk (S) isoform in lung and spleen.
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We thank Karl Fisher (University of Alberta) for helping us with the Western blot protocol, Thomas Turner (Laboratory Medicine and Pathology, University of Alberta) for technical support with pictures and also Bilan Li (Lovelace Respiratory Research Institute, Albuquerque) for technical assistance with cell transfection experiments. This work was supported by NIH 5 R 01 HL 069498 to Drs. A. D. Schreiber and A. D. Befus; by Sonderforschungsbereich SFB 633 to Dr. U. Steinhoff; by HL 071547 to Dr. K. Harrod.
Florentina Duta, Marina Ulanova and Daniel Seidel are joint first authors. Ulrich Steinhoff and A. Dean Befus are joint last authors.
M. Ulanova received a postdoctoral fellowship from the Canadian Society of Allergy and Clinical Immunology/Merck Frosst and from the Alberta Heritage Foundation for Medical Research.
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Duta, F., Ulanova, M., Seidel, D. et al. Differential expression of spleen tyrosine kinase Syk isoforms in tissues: effects of the microbial flora. Histochem Cell Biol 126, 495–505 (2006) doi:10.1007/s00418-006-0188-z
- Syk isoforms
- Epithelial cells