Characterization of P2Z purinergic receptors on phagocytic cells of the thymic reticulum in culture.
The thymic microenvironment is under intrinsic and extrinsic control circuits by several elements including hormones, neuropeptides, lymphokines, innervation and cell contact. P2 purinergic receptors have been described in a number of cells including macrophages, thymocytes, and other cells of the immune-inflammatory system. Here, we use the whole-cell patch-clamp technique and dye permeabilization assays to investigate the presence of ionic channels and purinergic receptors in one microenvironmental thymic component, namely the phagocytic cell of the thymic reticulum. At holding potentials ranging from -30 to -60 mV, applications of extracellular ATP in the vicinity of the cell membrane induce a transient and fast-activating inward current followed in most cells by an outward current. The whole event lasts 5-20 s. The inward current has a reversal potential close to 0 mV and the outward current can be ascribed to a Ca2+ -dependent K+ conductance. Both currents are inhibited by Mg2+, suggesting that the phenomenon is mediated by ATP4-. ATP-gamma-S can also induce both inward and outward currents. Exposure of phagocytic cells of the thymic reticulum to 5 mM ATP for 10 min induced permeabilization to lucifer yellow but not to the larger dyes trypan blue and rhodamine-dextran, suggesting a molecular weight cut-off smaller than 900. These observations lead us to conclude that phagocytic cells of the thymic reticulum express P2Z purinergic receptors that can mobilize Ca2+, induce the opening of ionic channels and permeabilize the cell membrane.
Coutinho-Silva
R
R
Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil.
Alves
L A
LA
de Carvalho
A C
AC
Savino
W
W
Persechini
P M
PM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D005456:Fluorescent Dyes; D008564:Membrane Potentials; D051379:Mice; D008807:Mice, Inbred BALB C; D010586:Phagocytes / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058486:Receptors, Purinergic P2X7; D015398:Signal Transduction; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism* / Q000502:physiology
D005456:Fluorescent Dyes; C546469:P2rx7 protein, mouse; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7; D000255:Adenosine Triphosphate