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Missense variants of hyperpolarization‐activated, cyclic nucleotide‐gated (HCN) ion channels cause variable phenotypes, ranging from mild generalized epilepsy to developmental and epileptic encephalopathy (DEE). Although variants of HCN1 are an established cause of DEE, those of HCN2 have been reported in generalized epilepsies. Here we describe the first case of DEE caused by the novel de novo heterozygous missense variant c.1379G>A (p.G460D) of HCN2. Functional characterization in transfected HEK293 cells and neonatal rat cortical neurons revealed that HCN2 p.G460D currents were strongly reduced compared to wild‐type, consistent with a dominant negative loss‐of‐function effect. Immunofluorescence staining showed that mutant channels are retained within the cell and do not reach the membrane. Moreover, mutant HCN2 also affect HCN1 channels, by reducing the Ih current expressed by the HCN1‐HCN2 heteromers. Due to the persistence of frequent seizures despite pharmacological polytherapy, the patient was treated with a ketogenic diet, with a significant and long‐lasting reduction of episodes. In vitro experiments conducted in a ketogenic environment demonstrated that the clinical improvement observed with this dietary regimen was not mediated by a direct action on HCN2 activity. These results expand the clinical spectrum related to HCN2 channelopathies, further broadening our understanding of the pathogenesis of DEE. [ABSTRACT FROM AUTHOR]

Aims: Nfix is a transcription factor belonging to the Nuclear Factor I (NFI) family comprising four members (Nfia, b, c, x). Nfix plays important roles in the development and function of several organs. In muscle development, Nfix controls the switch from embryonic to fetal myogenesis by promoting fast twitching fibres. In the adult muscle, following injury, lack of Nfix impairs regeneration, inducing higher content of slow‐twitching fibres. Nfix is expressed also in the heart, but its function has been never investigated before. We studied Nfix role in this organ. Methods: Using Nfix‐null and wild type (WT) mice we analyzed: (1) the expression pattern of Nfix during development by qPCR and (2) the functional alterations caused by its absence, by in vivo telemetry and in vitro patch clamp analysis. Results and Conclusions: Nfix expression start in the heart from E12.5. Adult hearts of Nfix‐null mice show a hearts morphology and sarcomeric proteins expression similar to WT. However, Nfix‐null animals show tachycardia that derives form an intrinsic higher beating rate of the sinus node (SAN). Molecular and functional analysis revealed that sinoatrial cells of Nfix‐null mice express a significantly larger L‐type calcium current (Cacna1d + Cacna1c). Interestingly, downregulation of Nfix by sh‐RNA in primary cultures of neonatal rat ventricular cardiomyocytes induced a similar increase in their spontaneous beating rate and in ICaL current. In conclusion, our data provide the first demonstration of a role of Nfix that, increasing the L‐type calcium current, modulates heart rate. [ABSTRACT FROM AUTHOR]

Ten years ago Yamanaka's lab identified a way to reprogram terminally differentiated cells to a pluripotent state, similar to that of embryonic stem cell. This procedure opened the road for the generation of postmitotic human cells, that have completely lost the replication potential. The initial excitement waned when it was observed that the cells produced by this method are somehow immature and do not resemble the adult phenotype. In the absence of cellular markers that recognize the various maturation steps of induced pluripotent stem cell-derived human cardiomyocytes, we propose to follow their maturation looking at their electrophysiological profile. For this reason, we are first reviewing the most common methods of differentiation, from the preliminary complex procedures to the newly-identified two-step protocols and, second, we report the electrical characteristics of the cells, through electrophysiological analysis of ionic currents that give rise to the action potential. We are aware that each protocol leads to the generation of different cardiomyocyte precursors, thus suggesting the need for a wider standardization. The identification of the electrophysiological characteristics of the cells could help in identifying the type and the maturation stage of the obtained cardiomyocyte, thus compensating for the lack of specific markers. Developmental Dynamics 245:1145-1158, 2016. © 2016 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Activation of the “funny” (pacemaker, If) current during the diastolic depolarization phase of an action potential is the main mechanism underlying spontaneous, rhythmic activity of cardiac pacemaker cells. In the past three decades, a wealth of evidence elucidating the function of the funny current in the generation and modulation of cardiac pacemaker activity has been gathered. The slope of early diastolic depolarization, and thus the heart rate, is controlled precisely by the degree of If activation during diastole. If is also accurately and rapidly modulated by changes of the cytosolic concentration of the second messenger cAMP, operated by the autonomous nervous system through β-adrenergic, mainly β2, and in the opposite way by muscarinic receptor, stimulation. Recently, novel in vivo data, both in animal models and humans, have been collected that confirm the key role of If in pacemaking. In particular, an inheritable point mutation in the cyclic nucleotide-binding domain of human HCN4, the main hyperpolarization-activated cyclic nucleotide (HCN) isoform contributing to native funny channels of the sinoatrial node, was shown to be associated with sinus bradycardia in a large family. Because of their properties, funny channels have long been a major target of classical pharmacological research and are now target of innovative gene/cell-based therapeutic approaches aimed to exploit their function in cardiac rate control. [ABSTRACT FROM AUTHOR]

Activation of the pacemaker (“funny,” If) current during diastole is the main process underlying generation of the diastolic depolarization and spontaneous activity of cardiac pacemaker cells. If modulation by autonomic transmitters is responsible for the chronotropic regulation of heart rate. Given its role in pacemaking, If has been a major target of investigation aimed to exploit its rate-controlling function in a clinical perspective. In this short review, we describe some of the most recent clinically relevant applications of the concept of If-based pacemaking. [ABSTRACT FROM AUTHOR]

The article discusses how the lipopolysaccharides (LPS) released from Gram-negative bacteria during sepsis can change the properties of the funny current . Topics covered include the heart rate (HR) modulation by the autonomic nervous system, the effects of LPS on HCN genes currents, and the important indicators of cardiovascular conditions.