Project description:External ionizing Irradiation of pregnant mice at E13.5 4 Gy, gene expression at E17.5 On E13.5, female C57Bl/6HN mice were irradiated (total body) at 0 Gy (sham) or 4 Gy using a Gamma Cell cesium irradiator (JL Shepherd, San Fernando, CA), with a dose rate of 70 cGy/min. The mice were monitored for 4 days, and on E17.5, they were sacrificed, and placentas were procured and processed for RNA extrtaction.
Project description:The mammalian brain is especially sensitive to ionizing radiation during development, as shown by the increased occurrence of mental retardation and small head size in children who were in utero exposed to ionizing radiation after the atomic bombings of Hiroshima and Nagasaki. These effects of prenatal irradiation can be mimicked by irradiation of mouse embryos during the organogenesis period. In order to better understand the early effects of ionizing radiation on the embryonic brain and immature neurons, we performed a microarray analysis on brains from mice irradiated with different doses (0.0, 0.1, 0.2, 0.5 and 1.0 Gy) at E11. RNA was extracted at either 2 or 24 h post-irradiation.
Project description:Here, we present the fetal mouse intestine data from the project \\"Comparison of human and mouse mesenchyme identifies common and unique aspects of intestinal patterning\\". Whole intestines were harvested from fetuses from timed pregnant matings for wildtype C57BL/6 mice (Jax strain #000664). Fetal stages were confirmed according to the Theiler staging chart (https://www.emouseatlas.org/emap/ema/staging_criteria/staging_criteria.html). Whole intestines (from the common bile duct through the cecum) were collected at key stages of development (E13.5, E14.5, E15.5, E16 and E17.5). Male and female intestines from each stage were pooled and dissociated to single cells for single cell RNA sequencing as previously described (Miller et al. 2020 Dev Cell). Specifically, E13.5 was 6 intestines, E14.5 was 5 intestines, E15.5 was 4 intestines, E16 was 3 intestines and E17.5 was 3 intestines.
Project description:Human embryonic stem cells (hESCs) present a novel platform for in vitro investigation of the early embryonic cellular response to ionizing radiation. Thus far, no study has analyzed the genome-wide transcriptional response to ionizing radiation in hESCs. In this study, we use Agilent microarrays to analyze the global gene expression changes in H9 hESCs after low (0.4 Gy), medium (2 Gy), and high (4 Gy) dose irradiation.
Project description:Tumor cell response to irradiation also depends on their microenvironment. Therefore ongoing investigation of three-dimensional (3D) cell culture models provide researchers with essential data studying and remodeling radiotherapeutic implications in cancer treatment. 3D culture models were shown to mimic in vivo cell microenvironment more accurately than the standard two-dimensional cell monolayer (2D) cultures. Growing evidence suggests that 2D and 3D cultured cell gene expression pattern discrepancies following irradiation is highly dependent on cell-ECM interactions. It has been shown that laminin-rich-extracellular matrix (lr-ECM) used in 3D cultures not only alters cancer cell phenotype and response to external stimuli but also affects their differentiation, migration and survivability. Thus, a change in these fundamental cell properties demands us to reconsider data previously collected using 2D in vitro models. RNA was harvested from two colorectal cancer cell lines cultivated under 3D cell culture conditions, 4h after treatment of single (2 Gy or 10 Gy) or fractionated (5x2 Gy) ionizing radiation dose.
Project description:Human embryonic stem cells (hESCs) present a novel platform for in vitro investigation of the early embryonic cellular response to ionizing radiation. Thus far, no study has analyzed the genome-wide transcriptional response to ionizing radiation in hESCs. In this study, we use Agilent microarrays to analyze the global gene expression changes in H9 hESCs after low (0.4 Gy), medium (2 Gy), and high (4 Gy) dose irradiation. Undifferentiated H9 hESCs were cultured on Matrigel in feeder-free conditions, and exposed to ionizing radiation at the indicated dosage (or control) from a Cesium-137 irradiator. Total RNA was isolated 24 hours after irradiation in the same feeder-free culture conditions. Experiment was repeated three times for each group, yielding a total of 12 distinct samples.
Project description:In this study, we measured mRNA and lncRNA profiles of gastric tissues following 0, 6 and 12 Gy irradiation. The stomach of C57 mice were exposed to 0, 6 and 12 Gy X-ray irradiation at a dose rate of 2 Gy/min. The mice were sacrificed 7 days after irradiation and gastric tissues were collected. RNA was extracted and quantified. mRNA and lncRNA profilings of each groups were analyzed by microarray.
Project description:Radiotherapy is one of the most common therapies for cancer. Approximately half of all cancer patients will receive radiotherapy at some point during treatment. Consequences of IR treatment are dose dependent and different sensitivity to IR of various types of cells is well established. To reduce the damage of IR to most sensitive cells of normal (noncancerous) tissue radiotherapy is administered as fractionated dose treatment applying radiation in ~2 Gy fractions every 24 hours, 5 times per week. However, during the therapy intrinsic and acquired tumor radioresistance may result in treatment failures. Comprehensive mechanisms of the resistance to irradiation as well as mechanisms of cellular response to fractionated dose IR remain unclear. Therefore, in the present study we evaluated global gene expression changes in murine Lewis lung carcinoma LLC1 cells following X-ray irradiation of single 2 Gy or 10 Gy and 2 Gy x 5 fractionated doses. Total RNA was harvested from mouse Lewis lung carcinoma cells 4h after treatment of single (2 Gy or 10 Gy) or fractionated (5x2 Gy) ionizing radiation dose.
Project description:We report whole transcriptome sequencing of the intraventricular septum of mouse heart 2 and 6 weeks after 25 Gy ionizing radiation. The objective of this experiment was to identify genes and pathways differentially regulated by ionizing radiation in the heart. Image-guided isocentric gamma irradiation was delivered to experimental groups under isoflurane anesthesia in a single, 25 Gy dose over approximately 15 minutes. Control group animals were experimental group littermates which received anesthesia and CT imaging but no radiation. RNA was isolated from intraventricular septum using TRIzol.
Project description:In this study we performed RNA sequencing to determine and compare the transcriptome of regulatory T cells (Tregs) present in the thymus, placenta, spleen and white adipose tissue (FAT) of pregnant mice. We further assessed how the expression of the receptor Rank in the thymic medullary epithelia affects the transcriptional program of these Treg populations. For the isolation of viable and bona fide Tregs, we first generated RankWt (wild type control mice) and RankΔFoxn1 (which lack expression of Rank in the thymic epithelia) and further crossed them to mice that express GFP from the Foxp3 promoter. Neuropilin1 was used to mark the thymic origin of the Tregs. Equal numbers of Tregs (280 cells) were sorted as CD45-CD8-CD4+GFP+Neuropilin1High cells from the thymus and placenta of RankWtFoxp3GFP/GFP and RankΔFoxn1Foxp3GFP/GFP pregnant females at E17.5 (both samples from the same female; n=4 females per genotype). Thymus Tregs from non-pregnant littermate female mice for each genotype cohort were also studied. To increase robustness, the 280 placental Tregs were purified from 5 individual placentas per pregnant female (56 Tregs/placenta). Our study is the first one to determine the transcriptome of thymus and placenta Tregs during pregnancy and reveals that, partially dependent on Rank expression in the thymic epithelia, placental-resident Tregs are molecularly distinct from thymic Tregs. In a separate experiment with different pregnant mice than those used for thymus and placenta analysis, VAT Tregs (20-30 cells) and splenic Tregs (150 cells) were sorted (both samples from the same female) by using the same mouse lines (Rank Wt and RankΔFoxn1), embryological days of analysis (E17.5) and sorting strategies.