Connexin expression and gap-junction-mediated cell interactions in an in vitro model of haemopoietic stroma

Abstract

In addition to the steady-state production of all blood cells, bone marrow can respond to an increased requirement for one or several cell lineages. The hormonal controls involved may act directly on blood cell progenitors or indirectly through modification of the haemopoietic environment. Intercellular gap junctions formed by connexins (Cx) provide direct communication among adjacent cells and the functional integration of multicellular systems. Since haemopoietic stroma is determinant for blood cell production, we have questioned whether gap-junction-dependent controls of haemopoiesis are sensitive to hormones and vitamins. We have analysed the expression, synthesis, cell distribution and formation of functional gap junctions in the murine bone-marrow stroma cell line S-17, and between stromal cells and blood cell progenitors. Nine Cxs were identified by reverse transcription/polymerase chain reaction, and only Cx43 by Western blot and immunofluorescence. All of the studied parameters were sensitive to intrinsic controls dependent upon the pattern of cell growth and modulated by exogenous controls mediated by retinol and steroids. Positive or negative modulation was specific for different Cxs. FACS analysis showed communication among the stromal cells and between stromal cells and myeloid (Mac1+) but not lymphoid (B220+) progenitors. Calcein transfer modulation did not correspond to the modulation of Cx43 expression and formation of connexons, suggesting the participation of other Cxs. Thus, functional gap junctions among haemopoietic stroma cells and between stroma and haematopoietic cells in the bone marrow may be modulated in response to hormonal stimuli, potentially controlling overall blood cell production.