Rationale: Cardiac pacemaker cells (PCs) in the sinoatrial node (SAN) have a distinct gene expression program that allows them to fire automatically and initiate the heartbeat. Although critical SAN transcription factors, including Isl1, Tbx3, and Shox2, have been identified, the cis-regulatory architecture that governs PC-specific gene expression is not understood, and discrete enhancers required for gene regulation in the SAN have not been identified. Objective: To define the epigenetic profile of PCs using comparative ATAC-seq and to identify novel enhancers involved in SAN gene regulation, development and function. Methods and Results: We used ATAC-seq on sorted neonatal mouse SAN to compare regions of accessible chromatin in PCs and right atrial cardiomyocytes. PC-enriched ATAC-seq peaks, representing candidate SAN regulatory elements, were located near established SAN genes and were enriched for distinct sets of TF binding sites. Among several novel SAN enhancers that were experimentally validated using transgenic mice, we identified a 2.9-kb regulatory element at the Isl1 locus that was active specifically in the cardiac inflow at E8.5 and throughout later SAN development and maturation. Deletion of this enhancer from the genome of mice resulted in SAN hypoplasia and sinus arrhythmias. The mouse SAN enhancer also directed reporter activity to the inflow tract in developing zebrafish hearts, demonstrating deep conservation of its upstream regulatory network. Finally, single nucleotide polymorphisms in the human genome that occur near the region syntenic to the mouse enhancer exhibit significant associations with resting heart rate in human populations. Conclusions: (1) PCs have distinct regions of accessible chromatin that correlate with their gene expression profile and contain novel SAN enhancers, (2) Cis-regulation of Isl1 specifically in the SAN depends upon a conserved SAN enhancer that regulates PC development and SAN function, and (3) a corresponding human ISL1 enhancer may regulate human SAN function.

Ecologically functional traits are the product of several, at times opposing, selective forces. Thus, ecomorphological patterns can be disrupted locally by biotic interactions, such as competition, and may not be consistent across lineages. Here, we studied the evolution of claws and toepads in relationship to macrohabitat (vegetation), use of structural microhabitat (perch height) and congeneric competition for two distantly related Lesser Antillean anole clades: the Anolis bimaculatus and Anolis roquet series. We collected univariate and geometric morphometric data from 254 individuals across 22 species to test the hypotheses that functional morphology should covary with both vegetation and perch height and that the presence of a competitor may disrupt such covariation. Our data showed predictable associations between morphology and macrohabitat on single-species islands but not when a congeneric competitor was present. The outcomes of competition differed between series, however. In the A. bimaculatus series, species with a sympatric congener diverged in claw and toepad traits consistent with functional predictions, whereas A. roquet series anoles showed either no association between habitat and morphology or the opposite pattern. Our results demonstrated that ecomorphological patterns across macrohabitats can be disrupted by competition-driven microhabitat partitioning and that specific morphological responses to similar ecological pressures can vary between lineages.

Cardiac pacemaker cells differentiate and functionally specialize early in embryonic development through activation of critical gene regulatory networks. In general, cellular specification and differentiation require that combinations of cell type-specific transcriptional regulators activate expression of key effector genes by binding to DNA regulatory elements including enhancers and promoters. However, because genomic DNA is tightly packaged by histones that must be covalently modified in order to render DNA regulatory elements and promoters accessible for transcription, the process of development and differentiation is intimately connected to the epigenetic regulation of chromatin accessibility. Although the difficulty of obtaining sufficient quantities of pure populations of pacemaker cells has limited progress in this field, the advent of low-input genomic technologies has the potential to catalyze a rapid growth of knowledge in this important area. The goal of this review is to outline the key transcriptional networks that control pacemaker cell development, with particular attention to our emerging understanding of how chromatin accessibility is modified and regulated during pacemaker cell differentiation. In addition, we will discuss the relevance of these findings to adult sinus node function, sinus node diseases, and origins of genetic variation in heart rhythm. Lastly, we will outline the current challenges facing this field and promising directions for future investigation.

The clearance of SARS-CoV-2 requires a multi-faceted immune response that is initiated by innate immune cells, with infection ultimately resolved by adaptive immune mechanisms. Induction of adaptive immunity to SARS-CoV-2 is dependent on the presentation of viral antigens on MHC II by professional antigen presenting cells such as dendritic cells and macrophages, to induce robust activation of CD4+ T cells. SARS-CoV-2 interferes with antigen presentation by downregulating MHC II on the antigen presenting cells of COVID-19 patients, but the molecular mechanism mediating this process is unelucidated. In this study, analysis of protein and gene expression in human antigen presenting cells reveals that the expression of MHC II is inhibited by the SARS-CoV-2 main protease, NSP5. Suppression of MHC II expression occurs via downregulation of the transcription factor CIITA, which is required for MHC II expression. This downregulation of CIITA is independent of NSP5's proteolytic activity, and rather, NSP5 delivers HDAC2 to the CIITA promoter via interactions with IRF3, Here, HDAC2 deacetylates and inactivates the CIITA promoter. This loss of CIITA expression prevents further expression of MHC II, with this suppression alleviated by ectopic expression of CIITA or knockdown of HDAC2. These results identify a novel mechanism by which SARS-CoV-2 can limit antigen presentation on MHC II, thereby delaying or weakening the subsequent adaptive immune response.

Color and pattern are often dynamic traits that change throughout an individual's lifetime. Still, long-term shifts in coloration have received limited attention. Dendrobatid poison frogs are a classical system in the study of color and pattern evolution in which both sexual selection and predation avoidance are thought to drive the evolution of color and pattern at the population and species level. Here, we highlight an overlooked axis of pattern diversity, within individual variation, using three species in the genus Dendrobates. We collected longitudinal photographs of individuals at the National Aquarium to test the hypothesis that patterns shift predictably throughout the lifetimes of individual frogs. In all three species, we found a consistent reduction in the relative area of aposematic color as individuals aged and that the rate of pattern shift did not differ between the sexes. Consequently, within individual variation in coloration may confound inferences from ecological studies that inherently assume individual pattern is static. Finally, we note that using simple and noninvasive photography protocols, animals in zoos and aquaria have the potential to deepen our understanding of how color and pattern change throughout the lifetimes of a wide range of species.