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).

Polarized hemichannel opening of pannexin 1/connexin 43 contributes to dysregulation of transport function in blood-brain barrier endothelial cells.

Dysregulation of blood-brain barrier (BBB) transport exacerbates brain damage in acute ischemic stroke. Here, we aimed to investigate the mechanism of this BBB transport dysregulation by studying the localization and function of pannexin (Px) and connexin (Cx) hemichannels in blood-brain barrier endothelial cells of rat (TR-BBB13 cells) and human (hCMEC/D3 cells) under acute ischemic stroke-mimicking oxygen/glucose deprivation (OGD) and extracellular Ca ([Ca])-free conditions. TR-BBB13 cells showed increased uptake of hemichannel-permeable sulforhodamine 101, and this increase was markedly inhibited by carbenoxolone, a hemichannel inhibitor.

Contribution of pannexin 1 and connexin 43 hemichannels to extracellular calcium-dependent transport dynamics in human blood-brain barrier endothelial cells.

Dysregulation of blood-brain barrier (BBB) transport function is thought to exacerbate neuronal damage in acute ischemic stroke. The purpose of this study was to clarify the characteristics of pannexin (Px) and/or connexin (Cx) hemichannel(s)-mediated transport of organic anions and cations in human BBB endothelial cell line hCMEC/D3 and to identify inhibitors of hemichannel opening in hCMEC/D3 cells in the absence of extracellular Ca(2+), a condition mimicking acute ischemic stroke. In the absence of extracellular Ca(2+), the cells showed increased uptake and efflux transport of organic ionic fluorescent dyes.