Dynamic expression of homeostatic ion channels in differentiated cortical astrocytes in vitro.

The capacity of astrocytes to adapt their biochemical and functional features upon physiological and pathological stimuli is a fundamental property at the basis of their ability to regulate the homeostasis of the central nervous system (CNS). It is well known that in primary cultured astrocytes, the expression of plasma membrane ion channels and transporters involved in homeostatic tasks does not closely reflect the pattern observed in vivo. The individuation of culture conditions that promote the expression of the ion channel array found in vivo is crucial when aiming at investigating the mechanisms underlying their dynamics upon various physiological and pathological stimuli. A chemically defined medium containing growth factors and hormones (G5) was previously shown to induce the growth, differentiation, and maturation of primary cultured astrocytes. Here we report that under these culture conditions, rat cortical astrocytes undergo robust morphological changes acquiring a multi-branched phenotype, which develops gradually during the 2-week period of culturing. The shape changes were paralleled by variations in passive membrane properties and background conductance owing to the differential temporal development of inwardly rectifying chloride (Cl-) and potassium (K+) currents. Confocal and immunoblot analyses showed that morphologically differentiated astrocytes displayed a large increase in the expression of the inward rectifier Cl- and K+ channels ClC-2 and Kir4.1, respectively, which are relevant ion channels in vivo. Finally, they exhibited a large diminution of the intermediate filaments glial fibrillary acidic protein (GFAP) and vimentin which are upregulated in reactive astrocytes in vivo. Taken together the data indicate that long-term culturing of cortical astrocytes in this chemical-defined medium promotes a quiescent functional phenotype. This culture model could aid to address the regulation of ion channel expression involved in CNS homeostasis in response to physiological and pathological challenges.

A new survival model based on ion channel genes for prognostic prediction in hepatocellular carcinoma.

Accumulating studies revealed the vital role of ion channels in cancers, but the prognosis role of ion channels in hepatocellular carcinoma (HCC) remains limited. Here, we developed and validated an ion channel signature for prognostic prediction of HCC patients. In total, 35 differential expressed ion channel genes (DEChannelGs) were identified in HCC and a novel ion channel risk model was established for HCC prognosis prediction using the TCGA cohort, which was validated using the ICGC cohort. Moreover, this risk model was an independent prognostic factor and was associated with the immune microenvironment in HCC. Finally, the mRNA and protein levels of ANO10 and CLCN2 were prominently up-regulated and were related to the poor prognosis of HCC patients. Taken together, these results indicated a novel ion channel risk model as a prognostic biomarker for HCC patients and provided further insight into its immunoregulatory mechanism in HCC progression.