Connexin-43 (Cx43) is the most abundant protein forming gap junction channels (GJCs) in cardiac ventricles. In multiple cardiac pathologies, including hypertrophy and heart failure, Cx43 is found remodeled at the lateral side of the intercalated discs of ventricular cardiomyocytes. Remodeling of Cx43 has been long linked to spontaneous ventricular arrhythmia, yet the mechanisms by which arrhythmias develop are still debated. Using a model of dystrophic cardiomyopathy, we previously showed that remodeled Cx43 function as aberrant hemichannels (non-forming GJCs) that alter cardiomyocyte excitability and, consequently, promote arrhythmias. Here, we aim to evaluate if opening of remodeled Cx43 can serve as a general mechanism to alter cardiac excitability independent of cellular dysfunction associated with a particular cardiomyopathy. To address this issue, we used a genetically modified Cx43 knock-in mouse (S3A) that promotes cardiac remodeling of Cx43 protein without apparent cardiac dysfunction. Importantly, when S3A mice were subjected to cardiac stress using the β-adrenergic agonist isoproterenol (Iso), they displayed acute and severe arrhythmias, which were not observed in WT mice. Pretreatment of S3A mice with the Cx43 hemichannel blocker, Gap19, prevented Iso-induced abnormal electrocardiographic behavior. At the cellular level, when compared with WT, Iso-treated S3A cardiomyocytes showed increased membrane permeability, greater plasma membrane depolarization, and Ca2+ overload, which likely caused prolonged action potentials, delayed after depolarizations, and triggered activity. All these cellular dysfunctions were also prevented by Cx43 hemichannel blockers. Our results support the notion that opening of remodeled Cx43 hemichannels, regardless of the type of cardiomyopathy, is sufficient to mediate cardiac-stress-induced arrhythmogenicity.

In arterial myocytes, the canonical function of voltage-gated CaV1.2 and KV2.1 channels is to induce myocyte contraction and relaxation through their responses to membrane depolarization, respectively. Paradoxically, KV2.1 also plays a sex-specific role by promoting the clustering and activity of CaV1.2 channels. However, the impact of KV2.1 protein organization on CaV1.2 function remains poorly understood. We discovered that KV2.1 forms micro-clusters, which can transform into large macro-clusters when a critical clustering site (S590) in the channel is phosphorylated in arterial myocytes. Notably, female myocytes exhibit greater phosphorylation of S590, and macro-cluster formation compared to males. Contrary to current models, the activity of KV2.1 channels seems unrelated to density or macro-clustering in arterial myocytes. Disrupting the KV2.1 clustering site (KV2.1S590A) eliminated KV2.1 macro-clustering and sex-specific differences in CaV1.2 cluster size and activity. We propose that the degree of KV2.1 clustering tunes CaV1.2 channel function in a sex-specific manner in arterial myocytes.

The expression of T-type voltage-dependent Ca 2+ channels (Cav3) has been previously observed in breast cancer, but their expression and subcellular localization were not evaluated in pre-neoplastic lesions. Therefore, this work aimed to evaluate protein expression and subcellular localization of T-type channel isoforms in human breast tissue samples. Protein expressions of Ca V 3.1, Ca V 3.2, and Ca V 3.3 were evaluated by immunohistochemistry in breast without alteration, in proliferative non-neoplastic lesions, and in neoplastic ductal epithelial lesions of the human breast. Ca V 3.1, Ca V 3.2, and Ca V 3.3 nuclear expressions were decreased in advanced stages of neoplastic transformation, whereas Ca V 3.1 and Ca V 3.2 cytoplasmic expression increased. Also, the decrease in nuclear expression was correlated with an increase in cytoplasmic expression for Ca V 3.1 isoform. The change in Ca V 3 protein expression and subcellular localization are consistent with the neoplastic transformation stages of mammary epithelial cells, evident in early neoplastic lesions, such as ductal carcinomas in situ . These results suggest a possible involvement of Ca V 3 in the carcinogenic processes and could be considered as a potential pharmacological target in new therapies for breast cancer treatment.

Background Intracellular Ca2+ levels can modulate several cellular functions, including proliferation and other processes found altered in neoplastic cells. Helping to maintain Ca2+ homeostasis, L-type voltage-dependent Ca2+ channels had its expression identified in neoplasias, including breast cancer. Invasive breast carcinoma of no special type, the most common classification of breast cancer, has ductal hyperplasia and ductal carcinoma in situ as its possible non-obligate precursors. This channel’s role in breast cancer development from these precursors has not been investigated. Evaluate protein expression and subcellular localization of CaV1.1, CaV1.2, and CaV1.3 in mammary epithelium without alteration and neoplastic and non-neoplastic ductal proliferative lesions through immunohistochemistry was the aim of this investigation. Methods In the present study, CaV1.1, CaV1.2, and CaV1.3 protein expression was evaluated by immunohistochemistry in breast without alteration and in proliferative non-neoplastic and neoplastic ductal epithelial lesions of the human breast. Results It was observed that CaV1.3 presented a reduction in nuclear expression at neoplastic lesions, in addition to an increase in cytoplasmic CaV1.1 expression. The analyses of membrane immunostaining showed that CaV1.2 and CaV1.3 had an increase of expression as the lesions progressed in the stages leading to invasive carcinomas. Conclusions Changes in protein expression and subcellular localization of these channels during the progression stages indicate that they may be involved in neoplastic transformation.