Project description:What methylation changes are occurring during seed development largely remains unknown. To uncover the possible role of DNA methylation during the transition from seed differentiation to maturation and dormany in soybean, we characterized the methylome of whole seeds representing the differentiation (GLOB stage), maturation (early- (EM), mid- (B1) and late- (AA1) maturation stages), and dormancy (DRY stage) phases of soybean seed development using Illumina sequencing. In addition, we characterized the methylome of the mid-maturation stage embryonic axis (B1-AX) to examine methylation differences, if any, between an embryonic region compared to the whole seed. Illumina sequencing of bisulfite-converted genomic DNA from globular stage (GLOB), early-maturation stage (EM), mid-maturation stage (B1), and late-maturation stage (AA1) seeds, dormancy stage (DRY) and mid-maturation embryonic axis (B1-AX).
Project description:In Brassicaceae and other species, the developing embryo develops in a spatially constrained environment, modulating embryo´s shape and size. It is unknown how embryonic cells adapt their growth and metabolism to the apparent mechanical cues, and in particular if embryo maturation is affected by physical constraints. To address this question, we performed topographical analysis (magnetic resonance imaging, infrared microspectroscopy, immunolabelling), metabolite, transcript and proteome profiling in developing seeds of Brassica napus. Here we describe how the imposition of mechanical stress during development causes a stimulation of oil and protein storage activity, characteristic of embryo maturation.
Project description:Seeds are comprised of three majors parts of distinct parental origin: the seed coat, embryo, and endosperm. The maternally-derived seed coat is important for nurturing and protecting the seeds during development. By contrast, the embryo and the endosperm are derived from a double fertilization event, where one sperm fertilizes the egg to form the diploid zygote and the other sperm fertilizes the central cell to form the triploid endosperm. Each seed parts undergo distinct developmental programs during seed development. What methylation changes occurring in the different seed parts, if any, remains unknown. To uncover the possible role of DNA methylation in different parts of the seed, we characterized the methylome of three major parts of an early maturation stage seed: seed coat, embryonic cotyledons, and embryonic axis using Illumina sequencing. Illumina sequencing of bisulfite-converted genomic DNA from three parts of an mid-maturation (B1) stage seed: seed coat (B1-SC), embryonic cotyledons (B1-COT), and embryonic axis (B1-AX).
Project description:Fibrotic disease is caused by persistently activated fibroblasts, known as myofibroblasts, that continuously deposit extracellular matrix and fail to de-activate after injury resolution. There are currently no treatments for fibrotic disease; our study addresses the mechanisms whereby myofibroblasts persist in fibrotic tissues such as diseased cardiac valves. We exploit photo-softening hydrogels as synthetic valve tissue mimics and valve fibroblasts as a model to study how stiffness controls pathological myofibroblast activation and their persistence. We show that persistent myofibroblasts have condensed chromatin structure with genome-wide alterations and that this is associated with stabilization of the actin cytoskeleton. Disconnecting the nucleus from the cytoskeleton prevents chromatin condensation and myofibroblast persistence. Notably, myofibroblasts in patients with aortic valve stenosis display a condensed chromatin structure compared to myofibroblasts in a healthy patient, similar to the difference observed between cultured persistent myofibroblasts and transient myofibroblasts. Collectively, our results reveal that nuclear mechanosensing leads to distinct chromatin signatures in persistent myofibroblasts and that this novel cellular mechanism is likely relevant to human fibrotic disease.
Project description:There are four major seed developmental phases in Arabidopsis seed development: morphogenesis, maturation, dormancy and germination. What methylation changes occurring in the different phases, if any, remains unknown. To uncover the possible role of DNA methylation in different parts of the seed, we characterized the methylome of four major seed developmental phases of Arabidopsis using Illumina sequencing: global stage (glob) and linear cotyledon stage (lcot) for morphogenesis phase; mature green stage (mg) and post mature green stage (pmg) for maturation phase; dry seed (dry) for dormancy phase; leaves (leaf) from 4 week plant for vegetative tissues. Illumina sequencing of bisulfite-converted genomic DNA from six seed developmental stages in Arabidopsis: global stage (glob), linear cotyledon stage (lcot), mature green stage (mg), post mature green stage (pmg), dry seed (dry) and leaves (leaf) from 4 week plant.
Project description:Matrix-derived biophysical cues are known to regulate the activation of fibroblasts and their subsequent transdifferentiation into myofibroblasts, but whether modulation of these signals can suppress fibrosis in intact tissues remains unclear, particularly in the cardiovascular system. We have employed single-cell/nucleus RNA sequencing of myofibroblasts differentiated from induced pluripotent stem cell-derived cardiac fibroblasts (iPSC-CFs), as well as mouse transverse aortic constriction (TAC) hearts to investigate whether modulation of matrix mechanosensing, in concert with TGF-beta inhibition, can potentiate the de-activation of fibrosis-promoting cardiac stromal populations in vitro and in vivo.
Project description:Purpose: The goal of our study is to compare two different ecotypes of Oryza sativa L., PHS-susceptible rice trait and PHS-resistant rice trait under three different maturation stages in rice seed embryo with profile of miRNA-seq. Methods: Oryza sativa. L miRNA profiles of two different ecotypes with 3 different maturation stages of rice seed embryo were generated by NGS, in duplicate, following Illumina NGS workflow. Results: We found the differentially expressed microRNAs between PHS-susceptible rice trait and PHS-resistant rice trait according to the three different seed maturation stages. Target transcripts of differentially expressed microRNAs have been predicted via psRNATarget web server, and a part of those target genes are likely to be regulated by microRNAs, affecting overall responses to heat stress and the regulation of seed dormancy during maturation. Conclusions: Our study represents the analysis of rice seed small RNAs, specifically microRNAs, under two different ecotypes, three different seed maturation stages in rice seed embryo. Our results show that microRNAs are involved in response to heat stress and the regulation of seed dormancy. This study will provide a foundation for understanding dynamics of seed dormancy during the seed development and overcoming pre-harvest sprouting.