Project description:Rationale: Despite the deleterious effects associated with elevated carbon dioxide (CO2), or hypercapnia, it has been hypothesized that CO2 can protect the lung from injury. However, the effects of chronic hypercapnia on the neonatal lung are unknown. Objectives: To determine whether chronic hypercapnia alters alveolar development and to identify genes that could potentially contribute to hypercapnia-mediated lung protection. Methods: Newborn mouse litters were exposed to 8% CO2, 12% CO2 or room air for 2 weeks. Lungs were excised and analyzed for morphometric alterations. Gene expression changes were assessed by microarray techniques and RT-PCR, and gene products by western blotting. Results: The alveolar walls of CO2-exposed mice appeared thinner than those of controls. In addition, genes from a variety of functional categories were differentially expressed with hypercapnia, including those involved with cell growth/maintenance, signal transduction, protein metabolism, ion transport, stress response and inflammation. In particular and of major interest, gene expression was increased for surfactant proteins (SP) A and D, epithelial Na+ channel, GATA binding protein 6 and fibroblast growth factor receptor 2. In addition, SP-A and SP-D protein expression was increased with hypercapnia. Conclusions: Our results lead us to conclude that: 1) There are potentially a number of gene families which may contribute to hypercapnia-mediated lung protection, and 2) up-regulation of SP-A and SP-D may play a role as anti-inflammatory or antioxidant agents. Based on our genomic results, the effects of CO2 depend on the level to which the lung is exposed. Total 5 arrays including 2 dye-swaps were performed for the 8% CO2 treated animals. Five individual animals were analyzed. Total 4 arrays including 2 dye-swaps were performed for the 12% CO2 treated animals. Four individual animals were analyzed.
Project description:Human HaCaT keratinocytes were cultured to confluence (day10). Differentiated confluent cells were irradiated at 0.5 Gy or 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap hybridization procedure 24 h after irradiation. Keywords = human keratinocytes, gamma irrdiation
Project description:Human keratinocytes were cultured to confluence (day 10). Differentiated confluent cells were irradiated at 0.5 and 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap procedure 24 h after irradiation. Keywords = Human keratinocytes, gamma irradiation
Project description:The keratinocyte cell line HaCaT was cultured for three days (proliferation) or for ten days (differentiation). RNA from cells at day3 was compared to RNA from cells at day10. The microarray hybridizations were performed in dye-swap procedure : RNA from cells at day3 was labeled with Cy3 (GSM4674, GSM4675, GSM4682, GSM4683) and then with cy5 (GSM4680, GSM4681). Keywords = cell density differentiation program in human keratinocytes
Project description:Human HaCaT keratinocytes were cultured to confluence (day10). Differentiated confluent cells were irradiated at 0.5 Gy or 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye swap hybridization procedure 3 h after irradiation. Keywords = Human keratinocytes, gamma irradiation
Project description:Huamn HaCaT keratinocytes were cultured to confluence (day10). Differentiated confluent cells were irradiated at 0.5 Gy or 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap hybridization procedure 3 h after irradiation. Keywords = Human keratinocytes, gamma irradiation Keywords: dose response
Project description:Chronic constant hypoxia (CCH), such as in pulmonary diseases or high altitude, and chronic intermittent hypoxia (CIH), such as in sleep apnea, can lead to major changes in the heart. The molecular mechanisms underlying these cardiac alterations are not well understood. We hypothesized that analysis on the changes in gene expression could help to delineate such mechanisms. In addition, the differences that can be anticipated between CCH and CIH could be potentially dissected. Current study used CCH and CIH mouse models combined with cDNA microarrays to determine the changes of gene expression in CCH or CIH mice heart. Keywords = heart Keywords = hypoxia Keywords = mouse Keywords = microarray Keywords: time-course
Project description:Hypoxia-induced cell injury has been related to multiple pathological conditions. In order to render hypoxia-sensitive cells and tissues resistant to low O2 environment, in this current study, we used Drosophila melanogaster as a model to dissect the mechanisms underlying hypoxia-tolerance. A D. melanogaster strain that lives perpetually in an extremely low-oxygen environment (4% O2, an oxygen level that is equivalent to that over about 4,000 m above Mt. Everest) was generated through laboratory selection pressure using a continuing reduction of O2 over many generations. This phenotype is genetically stable since selected flies, after several generations in room air, survive at this low O2 level. Gene expression profiling showed striking differences between tolerant and naïve flies, in larvae and adults, both quantitatively and qualitatively. Up-regulated genes in the tolerant flies included signal transduction pathways (e.g., Notch and Toll/Imd pathways), but metabolic genes were remarkably down-regulated in the larvae. Furthermore, a different allelic frequency and enzymatic activity of the triose phosphate isomerase (TPI) was present in the tolerant versus naïve flies. The transcriptional suppressor, hairy, was up-regulated in the microarrays and its binding elements were present in the regulatory region of the specifically down-regulated metabolic genes but not others, and mutations in hairy significantly reduced hypoxia tolerance. We conclude that, the hypoxia-selected flies: (a) altered their gene expression and genetic code, and (b) coordinated their metabolic suppression, especially during development, with hairy acting as a metabolic switch, thus playing a crucial role in hypoxia-tolerance. Keywords: genetic bases of hypoxia adaptation 27 isogenic D. melanogaster Lines were pooled and following long-term selection over generations with decreased oxygen level in the culture environment. The differences in gene expression were compared between adapted flies and generation matched naive controls by microarray. Pooled RNA samples from 3rd instar larvae of 27 parental lines were used as common reference.