Project description:This project aims to establish the embryonic anole lung as a model for investigating lung development and more general epithelial morphogenesis by generating a set of single-cell RNA-seq (scRNA-seq) data from late-stage embryonic lungs taken from brown anoles (Anolis sagrei). The anole lung, with its simple architecture, provides a novel tool for investigating signaling in a less complex respiratory system than the murine lung, and this data set would be a large advance in making this system more widely known and accesible among developmental biologists.
Project description:For the purpose of facilitating gene annotation, a transcriptome profile of the developing anole eye was used to identify coding and non-coding regions in the Anolis genome. RNA datasets were generated by collecting samples from three subregions of the anole posterior globe (central, temporal, and nasal) which was comprised of nerual retina, retinal pigmented epithelium, choriod, and scleral tissues. A total of five sample replicates were made. Each replicate incorporated pooled tissues collected from three stage 16.5 developing anole embryos.
Project description:The brown anole lizard, Anolis sagrei, is an emerging squamate model for developmental and functional genetic studies. To develop additional tools and resources for mechanistic studies of signaling pathways and cellular processes in A. sagrei, we established an in vitro system. Using this approach, we studied Hedgehog (Hh) signaling, one of the key developmental signaling pathways, which has evolved across the metazoa. We investigated Hh pathway-induced transcriptional changes in two evolutionarily distinct tetrapods: A. sagrei, and M. musculus, to identify the species-specific and evolutionarily shared responses. We report that ~45 % of genes induced as a response to Hh pathway activation in A. sagrei, are shared with M. musculus. To further increase the versatility of A. sagrei as a squamate model system for gene editing and genomic studies, we established and characterized a new immortalized A. sagrei embryonic fibroblast cell line ASEC-1. We performed whole-genome sequencing analysis to annotate the set of polymorphisms within this cell line. We conclude that transcriptome characterization of the ASEC-1 cell line would permit further investigations dissecting the complex biological and evolutionary aspects of Hh signaling.
Project description:In order to study the green anole dosage compensation mechanism we generated strand-specific RNA-seq libraries for a total of 186 samples from the green anole and other representative amniotes (human, mouse, opossum, platypus, chicken and xenopus). Moreover, in order to understand the dosage compensation mechanism of A. carolinensis, we generated ChIP-Seq data for the histone modification H4K16ac and compared levels of acetylation between males and females. Moreover, we reconstructed 7 Y-linked transcripts of Anolis carolinensis based on a male/female subtraction approach and validate these Y sequences using re-sequenced genomes.
