In vivo detection of intracellular signalling pathways in developing thymocytes

Michie, A.M. and Zúñiga-Pflücker, J.C. (2000) In vivo detection of intracellular signalling pathways in developing thymocytes. Developmental Immunology, 8(1), pp. 31-45.

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Information regarding the intracellular signaling processes that occur during the development of T cells has largely been obtained with the use of transgenic mouse models, which although providing invaluable information are time consuming and costly. To this end, we have developed a novel system that facilitates the in vivo analysis of signal transduction pathways during T-lymphocyte development. This approach uses reporter-plasmids for the detection of intracellular signals mediated by the mitogen-activated protein kinase or cyclic AMP-dependent protein kinase. Reporter-plasmids are transfected into thymocytes in fetal thymic organ culture by accelerated DNA/particle bombardment (gene gun), and the activation of a signaling pathway is determined in the form of a standard luciferase assay. Importantly, this powerful technique preserves the structural integrity of the thymus, and will provide an invaluable tool to study how thymocytes respond to normal environmental stimuli encountered during differentiation within the thymic milieu. Thus, this method allows for the monitoring of signals that occur in a biological time frame, such as during differentiation, and within the natural environment of differentiating cells.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Michie, Professor Alison
Authors: Michie, A.M., and Zúñiga-Pflücker, J.C.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:Developmental Immunology
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