Alves L A, Campos de Carvalho A C, Cirne Lima E O, Rocha e Souza C M, Dardenne M, Spray D C, Savino W
Department of Immunology, Institute Oswaldo Cruz, Janeiro, Brazil.
Eur J Immunol. 1995 Feb;25(2):431-7. doi: 10.1002/eji.1830250219.
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.
开展了一项多参数研究,以调查胸腺中尤其是胸腺微环境的主要成分胸腺上皮中通讯连接的存在情况及其可能的作用。通过原位和体外免疫组织化学标记、体外荧光染料注射以及双全细胞膜片钳实验,证实在人和小鼠胸腺上皮细胞(TEC)中存在由缝隙连接介导的直接细胞间通讯。此外,免疫印迹和Northern印迹研究均显示TEC中存在缝隙连接蛋白连接蛋白43及其mRNA。重要的是,我们发现,缝隙连接抑制剂辛醇可在体外特异性下调胸腺生成胸腺素所确定的胸腺内分泌活性。我们还研究了TEC与胸腺细胞之间缝隙连接的存在情况。在胸腺哺育细胞中,我们能够检测到细胞间通讯,尽管在特定时刻只有一小部分上皮/胸腺细胞对发生耦联。相比之下,在胸腺网状组织的培养吞噬细胞与各自形成花结的胸腺细胞之间未检测到细胞间通讯。我们认为,由连接蛋白43形成的缝隙连接可能代表胸腺内细胞通讯的一条新途径(且具有相当的细胞类型特异性),包括TEC/TEC以及可能的TEC/胸腺细胞相互作用。