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J Cell Sci ; 121(Pt 19): 3261-70, 2008 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-18782864

RESUMO

Macrophages express the P2X(7) receptor and other nucleotide (P2) receptors, and display the phenomenon of extracellular ATP (ATP(e))-induced P2X(7)-dependent membrane permeabilization, which occurs through a poorly understood mechanism. We used patch-clamp recordings, cytoplasmic Ca(2+) measurements and fluorescent dye uptake assays to compare P2X(7)-associated transport phenomena of macrophages and HEK-293 cells transfected with P2X(7) receptors (HEK-P2X(7) cells). Both cell types showed inward currents, increase of free cytoplasmic Ca(2+) concentration and the uptake of cationic dyes upon exposure to ATP(e), as previously described. However, in contrast to the macrophages, HEK-P2X(7) cells did not take up anionic dyes and did not display the 440 pS channels (Z pores) under cell-attached patch-clamping conditions. In addition, the transport mechanism of anionic dyes displayed by macrophages was also able to support dye efflux and, once activated at 37 degrees C, it remained active at 4 degrees C, whereas uptake of cationic dyes was temperature-dependent and unidirectional. Our results indicate that the mechanism of ATP(e)-induced dye uptake, usually called a ;permeabilization phenomenon' and associated with a ;permeabilization pore' can be ascribed to at least two distinct mechanisms in macrophages: a diffusional pathway, possibly associated with the 440 pS Z pores, and a cation uptake mechanism that is not diffusional and should be ascribed to an, as yet, unidentified transport mechanism.


Assuntos
Trifosfato de Adenosina/farmacologia , Ânions/metabolismo , Cátions/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Receptores Purinérgicos P2/metabolismo , Animais , Sinalização do Cálcio/efeitos dos fármacos , Linhagem Celular , Corantes/metabolismo , Difusão/efeitos dos fármacos , Etídio/metabolismo , Espaço Extracelular/efeitos dos fármacos , Espaço Extracelular/metabolismo , Humanos , Ativação do Canal Iônico/efeitos dos fármacos , Transporte de Íons/efeitos dos fármacos , Macrófagos/citologia , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Ratos , Receptores Purinérgicos P2X7 , Rodaminas
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