Sepsis induced channelopathy in skeletal muscles is associated with expression of non selective channels

Skeletal muscles (similar to 50% of the body weight) are affected during acute and late sepsis and represent one sepsis associate organ dysfunction. Cell membrane changes have been proposed to result from a channelopathy of yet unknown cause associated with mitochondrial dysfunction and muscle atrophy. We hypothesize that the channelopathy might be explained at least in part by the expression of non-selective channels. Here, this possibility was studied in a characterized mice model of late sepsis with evident skeletal muscle atrophy induced by cecal ligation and puncture (CLP). At day seven after CLP, skeletal myofibers were found to present de novo expression (immunofluorescence) of connexins 39, 43, and 45 and P2X7 receptor whereas pannexin1 did not show significant changes. These changes were associated with increased sarcolemma permeability (similar to 4 fold higher dye uptake assay), similar to 25% elevated in intracellular free-Ca2+ thorn concentration (FURA-2), activation of protein degradation via ubiquitin proteasome pathway (Murf and Atrogin 1 reactivity), moderate reduction in oxygen consumption not explained by changes in levels of relevant respiratory proteins, similar to 3 fold decreased mitochondrial membrane potential (MitoTracker Red CMXRos) and similar to 4 fold increased mitochondrial superoxide production (MitoSox). Since connexin hemichannels and P2X(7) receptors are permeable to ions and small molecules, it is likely that they are main protagonists in the channelopathy by reducing the electrochemical gradient across the cell membrane resulting in detrimental metabolic changes and muscular atrophy.Fondo Nacional de Ciencia y Tecnologia (FONDECYT) 1141092 1150291 FONDECYT 3160594 11160739 CONICYT/PAI 79140023 ICM-Economia Centro Interdisciplinario de Neurociencias de Valparaiso P09-022-

On biophysical properties and sensitivity to gap junction blockers of connexin 39 hemichannels expressed in hela cells

© 2017 Vargas, Cisterna, Saavedra-Leiva, Urrutia, Cea, Vielma, Gutierrez-Maldonado, Martin, Pareja-Barrueto, Escalona, Schmachtenberg, Lagos, Perez-Acle and Sáez. Although connexins (Cxs) are broadly expressed by cells of mammalian organisms, Cx39 has a very restricted pattern of expression and the biophysical properties of Cx39-based channels [hemichannels (HCs) and gap junction channels (GJCs)] remain largely unknown. Here, we used HeLa cells transfected with Cx39 (HeLa-Cx39 cells) in which intercellular electrical coupling was not detected, indicating the absence of GJCs. However, functional HCs were found on the surface of cells exposed to conditions known to increase the open probability of other Cx HCs (e.g., extracellular divalent cationic-free solution (DCFS), extracellular alkaline pH, mechanical stimulus and depolarization to positive membrane potentials). Cx39 HCs were blocked by some traditional Cx HC blockers, but not by others or a pannexin1 channel blocker. HeLa-Cx39 ce