Laser Micro‐dissection and Quantitative RT‐PCR as a Tool for Cell Identification

We have applied a laser micro‐dissection microscope followed by RT‐PCR analysis of micro‐dissected tissue to identify gene expression of various immune genes in the Atlantic salmon, Salmo salar. A major problem in teleost anatomy is the lack of specific cell markers to identify different leukocyte p...

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Bibliographic Details
Published in:Anatomia, Histologia, Embryologia
Main Authors: Koppang, E. O., Huang, Q., Hordvik, I., Landsverk, T., Press, C. McL., Bakke‐McKellep, A. M.
Format: Article in Journal/Newspaper
Language:English
Published: Wiley 2005
Subjects:
Atlantic salmon
Salmo salar
Online Access:http://dx.doi.org/10.1111/j.1439-0264.2005.00669_61.x
https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1439-0264.2005.00669_61.x
https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1439-0264.2005.00669_61.x
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Summary:We have applied a laser micro‐dissection microscope followed by RT‐PCR analysis of micro‐dissected tissue to identify gene expression of various immune genes in the Atlantic salmon, Salmo salar. A major problem in teleost anatomy is the lack of specific cell markers to identify different leukocyte populations. For instance, the involvement of T‐cells in inflammatory reactions is unknown. We used frozen sections from the gut of salmon fed to induce localized inflammatory reactions and micro‐dissected regions showing proliferative changes. RNA was extracted from the micro‐dissected specimens and subjected to quantitative RT‐PCR using primers and TaqMan probes for T‐cell receptor, MHC class II and elongation factor, which served as a house‐keeping gene. The results showed expression of T‐cell receptor, MHC class II and elongation factor. The expression of the elongation factor corresponded invariably to the area micro‐dissected tissue. In conclusion, T‐cells are present in teleost inflammatory reactions.

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