The Genomic Distribution and Function of Histone Variant HTZ-1 during C. elegans Embryogenesis

In all eukaryotes, histone variants are incorporated into a subset of nucleosomes to create functionally specialized regions of chromatin. One such variant, H2A.Z, replaces histone H2A and is required for development and viability in all animals tested to date. However, the function of H2A.Z in development remains unclear. Here, we use ChIP-chip, genetic mutation, RNAi, and immunofluorescence microscopy to interrogate the function of H2A.Z (HTZ-1) during embryogenesis in Caenorhabditis elegans, a key model of metazoan development. We find that HTZ-1 is expressed in every cell of the developing embryo and is essential for normal development. The sites of HTZ-1 incorporation during embryogenesis reveal a genome wrought by developmental processes. HTZ-1 is incorporated upstream of 23% of C. elegans genes. While these genes tend to be required for development and occupied by RNA polymerase II, HTZ-1 incorporation does not specify a stereotypic transcription program. The data also provide evidence for unexpectedly widespread independent regulation of genes within operons during development; in 37% of operons, HTZ-1 is incorporated upstream of internally encoded genes. Fewer sites of HTZ-1 incorporation occur on the X chromosome relative to autosomes, which our data suggest is due to a paucity of developmentally important genes on X, rather than a direct function for HTZ-1 in dosage compensation. Our experiments indicate that HTZ-1 functions in establishing or maintaining an essential chromatin state at promoters regulated dynamically during C. elegans embryogenesis

Genetics of germ cell development

The germ line represents a continuous cellular link between generations and between species, but the germ cells themselves develop in a specialized, organism-specific context. The model organisms Caenorhabditis elegans, Drosophila melanogaster and the mouse display striking similarities, as well as major differences, in the means by which they control germ cell development. Recent developments in genetic technologies allow a more detailed comparison of the germ cells of these three organisms than has previously been possible, shedding light not only on universal aspects of germline regulation, but also on the control of the pluripotent state in vivo and on the earliest steps of embryogenesis. Here, we highlight themes from the comparison of these three alternative strategies for navigating the fundamental cycle of sexual reproduction.Howard Hughes Medical Institute (Investigator

Decellularized vascular grafts

Cardiovascular disease is one of the main causes of mortality and morbidity worldwide. The “gold standard” for the replacement/repair of diseased blood vessels is substitution with autologous vessels. However, multiple surgical procedures limit the availability of autologous vessels, whereas synthetic grafts have been reported to demonstrate poor patency rates, especially for small-caliber vascular reconstructions. Decellularization of native vascular or non-vascular tissues for vascular scaffold development has gained significant attention in the past 20 years. A variety of decellularization techniques have been described and employed to achieve effective immunogenic agent removal from the developed vascular scaffold. At the same time, the decellularization must not impair the extracellular matrix (ECM) composition, structure, and mechanical properties of the graft in order to ensure long-term functionality in vivo. The aim of this chapter was to review the various decellularization treatments that have been reported in the literature for the development of decellularized vascular scaffolds