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Published on: July 16, 2013
Functional gap junctions in thymic epithelial cells are formed by connexin 43
L A Alves1, A C Campos de Carvalho, E O Cirne Lima
1Department of Immunology, Institute Oswaldo Cruz, Janeiro, Brazil.
Insights
Gap junctions facilitate cell communication in the thymus, particularly between thymic epithelial cells (TEC). Connexin 43 mediates this communication, influencing thymic endocrine activity and potentially TEC-thymocyte interactions.
Area of Science:
- Immunology
- Cell Biology
- Histology
Background:
- The thymus microenvironment is crucial for T-cell development.
- Thymic epithelial cells (TEC) form the primary structure of this microenvironment.
- The role of intercellular communication, specifically gap junctions, in TEC function is not well understood.
Purpose of the Study:
- To investigate the presence and function of gap junctions in the thymus.
- To determine the role of gap junctions in thymic epithelial cell (TEC) communication.
- To explore potential TEC-thymocyte communication via gap junctions.
Main Methods:
- In situ and in vitro immunohistochemical labeling of human and murine thymic tissue.
- In vitro fluorochrome injection and double whole-cell patch clamp experiments on TEC.
- Immuno- and Northern blot analyses for connexin 43 expression.
- In vitro experiments using a gap junction inhibitor (octanol) to assess thymic endocrine activity.
- Analysis of gap junction presence between TEC and thymocytes in thymic nurse cells and phagocytic cells.
Main Results:
- Gap junctions were confirmed in human and murine TEC, indicating direct cell-cell communication.
- Connexin 43 protein and mRNA were detected in TEC.
- Thymic endocrine activity, measured by thymulin production, was modulated by a gap junction inhibitor.
- Minor cell-cell communication was observed between TEC and thymocytes in thymic nurse cells.
- No intercellular communication was detected between phagocytic cells and thymocytes.
Conclusions:
- Gap junctions, mediated by connexin 43, represent a novel pathway for intrathymic cellular communication.
- This communication occurs between TEC and potentially between TEC and thymocytes.
- Gap junction activity influences thymic endocrine function.
Abstract:
A multiparametric study was carried out to investigate the presence and possible role of communicating junctions in the thymus, particularly in the thymic epithelium, the major component of the thymic microenvironment. The presence of direct cell-cell communication mediated by gap junctions was demonstrated in human and murine thymic epithelial cells (TEC) by means of in situ and in vitro immunohistochemical labeling as well as in vitro fluorochrome injection and double whole-cell patch clamp experiments. Moreover, both immuno- and Northern blot studies revealed that the gap junction protein connexin 43 and its mRNA were present in TEC. Importantly, we showed that thymic endocrine activity, as ascertained by thymulin production, could be specifically down-modulated in vitro by a gap junction inhibitor, octanol. We also investigated the existence of gap junctions between TEC and thymocytes. In thymic nurse cells we were able to detect cell-cell communication, although only a minor percentage of epithelial/thymocyte pairs were coupled in a given moment. In contrast, intercellular communication was not detected between cultured phagocytic cells of the thymic reticulum and the respective rosetting thymocytes. We suggest that gap junctions formed by connexin 43 may represent a novel (and rather cell type-specific) pathway for intrathymic cellular communication, including TEC/TEC as well as possible TEC/thymocyte interactions.
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