Tugging on the heartstrings: determining the function of Tbx5 in early cardiac development

2012 Spring.Includes bibliographical references.During cardiac morphogenesis, the vertebrate heart acquires a characteristic three dimensional shape well-suited for efficient function. The morphology of the developing cardiac organ reflects a series of changes in the cardiomyocytes themselves, which must become specified, migrate, proliferate, grow in size, alter their shape and adhesive properties, and develop ultrastructure, among other differentiated characteristics. Mutation of the T-box transcription factor tbx5 leads to embryonic lethal cardiac phenotypes and forelimb malformations in vertebrate models. Haploinsufficiency of Tbx5 results in Holt-Oram Syndrome (HOS), a human congenital disease characterized by cardiac and forelimb defects. Homozygous mutation of zebrafish tbx5a in heartstrings (hst) embryos also leads to lethal defects in cardiac looping morphogenesis and prevents initiation of pectoral fin formation. Here I describe a new hst mutant allele (tbx5as296) which encodes a premature stop codon within the tbx5a T-box region, a location likely to generate a full loss-of-function allele. Data from comparative genetics and immunoblot analyses indicate that both alleles are null. I find that mutants completely lacking Tbx5a generated normal cardiomyocyte numbers in early chamber morphogenesis stages. Moreover, in situ hybridization data and functional assays support the idea that venous differentiation is not seriously impaired in zebrafish mutants, in contrast to mouse. However, cardiac cell size was significantly smaller in both chambers of tbx5a mutants. Hearts stalled early in the process of cardiac looping, but cell shape changes associated with chamber ballooning surprisingly still occurred. These studies point to a critical role for Tbx5a in growth-related aspects of cardiac differentiation, and suggest that morphologic events of cardiac looping morphogenesis and chamber ballooning are genetically separable. A second zebrafish tbx5 paralog was recently described, termed tbx5b, which showed a lower amount of sequence conservation than is typical for a T-box gene. Based on overlapping expression patterns within the embryonic heart, I hypothesized that functional redundancy between tbx5a and tbx5b might reduce the severity of cardiac phenotypes for tbx5a mutant embryos. I here report that the cardiac phenotypes in tbx5b-depleted fish were similar, but not identical, to those of homozygous tbx5a mutants. In addition, tbx5b-depletion led to defects in the timing and morphogenesis of pectoral fin outgrowth. Somewhat surprisingly, simultaneous depletion of both Tbx5 gene paralogs did not lead to more severe cardiac phenotypes, and injection of wild-type mRNA was not sufficient to cross-rescue the phenotypes of the paralogous gene. In the heart, tbx5a and tbx5b appear to have related essential functions that are nevertheless independently required. In the fin, tbx5a alone was required for fin bud initiation, but both genes are independently required for patterning and morphogenesis. Therefore, this work demonstrates a functional divergence between the two zebrafish tbx5 paralogs

Oligodendrocytes Are a Major Target of the Toxicity of Spongiogenic Murine Retroviruses

The neurovirulent retroviruses FrCasE and Moloney MLV-ts1 cause noninflammatory spongiform neurodegeneration in mice, manifested clinically by progressive spasticity and paralysis. Neurons have been thought to be the primary target of toxicity of these viruses. However the neurons themselves appear not to be infected, and the possible indirect mechanisms driving the neuronal toxicity have remained enigmatic. Here we have re-examined the cells that are damaged by these viruses, using lineage-specific markers. Surprisingly, these cells expressed the basic helix-loop-helix transcription factor Olig2, placing them in the oligodendrocyte lineage. Olig2(+) cells were found to be infected, and many of these cells exhibited focal cytoplasmic vacuolation, suggesting that infection by spongiogenic retroviruses is directly toxic to these cells. As cytoplasmic vacuolation progressed, however, signs of viral protein expression appeared to wane, although residual viral RNA was detectable by in situ hybridization. Cells with the most advanced cytoplasmic effacement expressed the C/EBP-homologous protein (CHOP). This protein is up-regulated as a late event in a cellular response termed the integrated stress response. This observation may link the cellular pathology observed in the brain with cellular stress responses known to be induced by these viruses. The relevance of these observations to oligodendropathy in humans is discussed

Multimodal surveillance of SARS-CoV-2 at a university enables development of a robust outbreak response framework

AbstractUniversities are particularly vulnerable to infectious disease outbreaks and are also ideal environments to study transmission dynamics and evaluate mitigation and surveillance measures when outbreaks occur. Here, we introduce a SARS-CoV-2 surveillance and response framework based on high-resolution, multimodal data collected during the 2020-2021 academic year at Colorado Mesa University. We analyzed epidemiological and sociobehavioral data (demographics, contact tracing, and wifi-based co-location data) alongside pathogen surveillance data (wastewater, random, and reflexive diagnostic testing; and viral genomic sequencing of wastewater and clinical specimens) to characterize outbreak dynamics and inform policy decisions. We quantified group attributes that increased disease risk, and highlighted parallels between traditional and wifi-based contact tracing. We additionally used clinical and environmental viral sequencing to identify cryptic transmission, cluster overdispersion, and novel lineages or mutations. Ultimately, we used distinct data types to identify information that may help shape institutional policy and to develop a model of pathogen surveillance suitable for the future of outbreak preparedness.</jats:p

Multimodal surveillance of SARS-CoV-2 at a university enables development of a robust outbreak response framework

Background: Universities are vulnerable to infectious disease outbreaks, making them ideal environments to study transmission dynamics and evaluate mitigation and surveillance measures. Here, we analyze multimodal COVID-19-associated data collected during the 2020-2021 academic year at Colorado Mesa University and introduce a SARS-CoV-2 surveillance and response framework. Methods: We analyzed epidemiological and sociobehavioral data (demographics, contact tracing, and WiFi-based co-location data) alongside pathogen surveillance data (wastewater and diagnostic testing, and viral genomic sequencing of wastewater and clinical specimens) to characterize outbreak dynamics and inform policy. We applied relative risk, multiple linear regression, and social network assortativity to identify attributes or behaviors associated with contracting SARS-CoV-2. To characterize SARS-CoV-2 transmission, we used viral sequencing, phylogenomic tools, and functional assays. Findings: Athletes, particularly those on high-contact teams, had the highest risk of testing positive. On average, individuals who tested positive had more contacts and longer interaction durations than individuals who never tested positive. The distribution of contacts per individual was overdispersed, although not as overdispersed as the distribution of phylogenomic descendants. Corroboration via technical replicates was essential for identification of wastewater mutations. Conclusions: Based on our findings, we formulate a framework that combines tools into an integrated disease surveillance program that can be implemented in other congregate settings with limited resources. Funding: This work was supported by the National Science Foundation, the Hertz Foundation, the National Institutes of Health, the Centers for Disease Control and Prevention, the Massachusetts Consortium on Pathogen Readiness, the Howard Hughes Medical Institute, the Flu Lab, and the Audacious Project