Distinct Transport Properties of Human Pannexin 1 and Connexin 32 Hemichannels.

Pannexin (Px) and connexin (Cx) hemichannels mediate bidirectional membrane transport in response to various stimuli and are involved in drug efficacy and toxicity. The purpose of the present study was to clarify in detail the transport characteristics of Px1 and Cx32 hemichannels by establishing transport assay systems using human Px1- and P2RX7 receptor-overexpressing HEK293 cells (Px1/P2RX7/HEK293) and Cx32-overexpressing HEK293 cells (Cx32/HEK293), in which P2RX7 and an extracellular Ca-depleted condition serve as the opening trigger, respectively. Uptake of the cationic fluorescent dye propidium iodide (PI) was significantly increased in Px1/P2RX7/HEK293 cells compared to that in mock cells, whereas there was no significant uptake of the anionic fluorescent dye sulforhodamine 101 (SR101). Uptake of [H]cholesterol by Px1/P2RX7/HEK293 cells was significantly decreased, whereas that of [H]taurine was not, compared to mock cells. On the other hand, uptakes of PI and SR-101 by Cx32/HEK293 cells were both significantly increased compared to mock cells. The PI uptake by Cx32/HEK293 cells was significantly inhibited by thioacetamide, acetaminophen, and N-acetyl-p-benzoquinoneimine. Cellular uptake of [H]cholesterol was significantly increased in Cx32/HEK293 cells and that of [H]taurine was significantly decreased. These results support the idea that Px1 and Cx32 hemichannels have distinct substrate recognition specificities and transport directions.