Project description:Mesophotic coral reefs have been proposed as refugia for corals, providing shelter and larval propagules for shallow-water reefs that are disproportionately challenged by global climate change and local anthropogenic stressors. Yet, knowledge of the capacity of coral larvae to adjust to different depth environments is still limited. In this study, planulae of the reef-building coral Stylophora pistillata from 5-8 and 40-44 m depth in the Gulf of Aqaba were tested in a long-term in situ translocation experiment for their ability to settle and acclimate to reciprocal depth conditions. We assessed survival rates, photochemical, physiological and morphological characteristics, as well as gene expression variations in juveniles grown at different depths, comparing them to non-translocated adults, juveniles and planulae. We found high mortality rates among mesophotic-origin planulae, irrespective of translocation depth. Gene expression patterns suggested that deep planulae lacked settlement competency and experienced increased developmental stress upon release. Symbiont photochemical acclimation to depth occurred rapidly within 8 days, with symbiont populations showing changes in photochemical traits but no symbiont species shuffling between deep and shallow juveniles. In contrast, coral host physiological and morphological acclimation were less evident. We observed minimal overlap in gene expression patterns between different life stages and depths, indicating that gene expression significantly depends on life stage. The study also identified a set of DEGs associated with initial stress responses following translocation, lingering stress response, and environmental effects of depth. In conclusion, though our data reveal rapid symbiont acclimation, host acclimation to match deep coral phenotypes was incomplete within 60 days for planulae translocated to different depths. These results have implications for understanding the ecological significance of mesophotic reefs as potential larval sources in the face of environmental stressors.
Project description:We designed a new specific mRNA microarray targeting a subset of genes (748) of the diazotrophs Richelia intracellularis and Calothrix rhizosoleniae (genomes RintRC01, RintHH01, RintHM01 and CalSC01) which associate with diatom hosts. The aim was to be able to describe the gene expressions of genes related to several metabolic pathways and how they possibly differed between the closely related strains based on environment and host association. To better understand how the different environments might affect gene expressions, the samples were taken in depth profiles, at night and day, during a cruise in the South China Sea.
Project description:Recent advances in RNA sequencing (RNA-Seq) have enabled the discovery of novel transcriptomic variations that are not possible with traditional microarray-based methods. Tissue and cell specific transcriptome changes during pathophysiological stress, in disease cases versus controls and in response to therapies are of particular interest to investigators studying cardiometabolic diseases. Thus, knowledge on the relationships between sequencing depth and detection of transcriptomic variation is needed for designing RNA-Seq experiments and for interpreting results of analyses. Using deeply sequenced RNA-Seq data derived from adipose of a healthy individual before and after systemic administration of endotoxin (LPS), we investigated the sequencing depths needed for studies of gene expression and alternative splicing (AS). We found that to detect expressed genes and AS events, ~100 million (M) filtered reads were needed. However, the requirement on sequencing depth for the detection of LPS modulated differential expression (DE) and differential alternative splicing (DAS) was much higher. To detect 80% of events, ~300M filtered reads were needed for DE analysis whereas at least 400M filtered reads were necessary for detecting DAS. Although the majority of expressed genes and AS events can be detected with modest sequencing depths (~100M filtered reads), the estimated gene expression levels and exon/intron inclusion levels were less accurate. We report the first study that evaluates the relationship between RNA-Seq depth and the ability to detect DE and DAS in human adipose. Our results suggest that a much higher sequencing depth is needed to reliably identify DAS events than for DE genes. Random sampling the RNA-seq data in different depth for gene and alternative-splicing analysis
Project description:We have developed an endovascular catheter-based device for sampling of the endothelium, intended for RNA-seq analysis. With an outer diameter of 0.23 mm it is capable of reaching peripheral vessels for endothelial sampling, and favourably compares to other approaches with stent retrievers or similar tools. Here, we have collected samples from liver and kidney vessels from swine and performed RNA-sequencing to validate the sample quality.