Project description:Hyperoxia has a potential to alter DNA methylation status. We assessed the effect of long term hyperoxia in mouse lung tissue. A total of 24 mice were randomized to hyperoxia (85% O2; 12 animals) or normoxia (21% O2; 12 animals) for 14 days continued with normoxia conditions for all animals for the subsequent 14 days. All mice had free access to food and water and were kept under standard conditions in A-Chambers (O2 – monitor ProOX110, CO2 – monitor ProCO2 P120, BioSpherix). The animals were sacrificed on day 28. Lung tissue was harvested on day 28 after euthanasia with a zolazepam/tiletamine/xylazine/fentanyl cocktail. Tissues samples were snap-frozen in liquid nitrogen immediately after cessation of circulation for the subsequent analysis. Subsequently, DNA methylation profiles in lung tissue were compared by means of methylation microarrays between both groups. We used the Genomic Workbench software (Agilent) to assess the mean methylation status of each DNA fragment (array probe) in each group. We first calculated the normalised, combined Z-scores, representing summation of the left and right Gaussian Z-scores and reflecting the location of a probe log-ratio value in relation to the Gaussian distribution of probes. A strong positive value of the combined Z-score means that a given probe is methylated and strong negative value means that it is unmethylated. Next, we compared the mean combined Z-scores (average methylation patterns) of the probes between hyperoxia and normoxia groups.
Project description:Newborn mouse pups were exposed either to normoxia (21% O2) or to hyperoxia (85% O2) since the day of birth. Lungs were harvested at P2.5, P3.5, P5.5 and P14.5 and homogenized for a downstream microRNA (miR) microarray in order to compare the differential expression of miRs between normoxia and hyperoxia group. Several miRs appeared to be dysregulated, mainly and with a higher significance, at P5.5 and P14.5.
Project description:Lung tissue from normoxia- or hyperoxia-treated control or SCNN1B overexpression mice were processed with a redox proteomics workflow and analyzed by LC-MS/MS with TMT based quantification. Data was searched with MS-GF+ using PNNL's DMS Processing pipeline.
Project description:Akap1 KO and Wt mice were exposed to normoxia or hyperoxia for 48h. Total RNA was extracted from lungs of Wt Normoxia (n=3), Wt hyperoxia (n=3), Akap1 KO (n=3) and Akap1 hyperoxia (n=3) mice. RNA-sequencing was carried out followed by differential expression of genes in the following groups. Wt Normoxia vs Wt Hyperoxia, Akap1 KO Normoxia versus Akap1 KO Hyperoxia, Wt Normoxia versus Akap1 KO Normoxia and Wt Hperoxia versus Akap1 Hyperoxia.
Project description:Extremely preterm infants are often treated with supraphysiological oxygen which contributes to the development of bronchopulmonary dysplasia (BPD). These same infants exhibit compromised antioxidant capacities due in part to selenium (Se) deficiency. The present study was designed to develop a perinatal Se deficiency mouse model, identify the effects of newborn hyperoxia exposure, and explore alternative pathways affected by Se deficiency (SeD) that would contribute to impaired lung development. Se deficient breeding pairs were generated, once pups were born, they were exposed to room air or 85% O2 for 14 d. Survival, antioxidant and Nrf2-regulated protein expression, and RNA seq analyses were performed. Greater than 40% mortality was observed in Se deficient (SeD), hyperoxia exposed pups. Surviving SeD pups had greater lung growth deficits than Se sufficient (SeS) pups exposed to hyperoxia. Gpx2 and 4 protein and Gpx activity were significantly decreased in SeD pups. Nrf2-regulated proteins, NQO1 and Gclc were increased in the setting of Se deficiency and hyperoxia exposure. RNA seq revealed significant decreases in the Wnt/-catenin and Notch pathways. Se is a biologically relevant modulator of perinatal lung development and antioxidant responses, especially in the context of hyperoxia exposure. RNA seq implicates pathways essential for normal lung development are dysregulated by Se deficiency.
Project description:Our previous data obtained by using immunohistochemistry showed, that Fgf10+/- (50% Fgf10 expression compared to WT) in hyperoxic condition at postnatal day 3 (P3) compared to WT has less vessel count in the lung and less muscularization of small capillaries in the lung. Furthermore, Fgf10+/- showed a drastic increase in mortality upon hyperoxic lung injury. Main question to be answer by this experiment is as followed: Does Fgf10+/- mice after hyperoxia from P0-P3 show different expression profiles at P3 compared to WT? To adress this question we harvest lungs at P3 from WT and Fgf10+/- after hyperoxia treatment from P0-P3. For this the mice were sacrificed by Ketamin/ Dormitor ip, lungs were perfused transcardiac with PBS and directly frozen in liquid nitrogen.
Project description:In this study the microRNA expression of primary murine (C57BL/6) lung alveolar type II cells belonging to four different conditions was analyzed. Effects of hyperoxia 24 hours group were compared to the normoxia 24 hours and effects of the hyperoxia 6 hours group were compared to the normoxia 6 hours.
Project description:Purpose: To study the effect of hyperoxia on lung cancer cells, H1299 cells were cultured in 60% O2 for 12 h and 24 h, and RNA-seq was performed.
Project description:In this study the gene expression of primary murine (C57BL/6) lung alveolar type II cells belonging to four different conditions was analyzed. Effects of hyperoxia 24 hours group (group 2) were compared to the normoxia 24 hours (group 1) and effects of the hyperoxia 6 hours group (group 4) were compared to the normoxia 6 hours (group 3)
Project description:In this experiment 2 day old baby mice were exposed to hyperoxia (75% O2) continuously for 14 days. Control baby mice were housed in room air (normoxia). Plasma and bronchoalveolar lavage fluid (BALF) were harvested after 14 days of exposure (on Day of life 16)