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Status |
Public on Apr 17, 2019 |
Title |
In utero exposure to diesel exhaust particulates is associated with alterations of the neonatal cardiomyocyte transcriptome, metabolic parturbation and a global decrease in DNA methylation. [RNA-Seq] |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Developmental exposure to particulate matter air pollution is harmful to cardiovascular health, but the mechanisms by which this exposure mediates susceptibility to heart disease is poorly understood. We have previously shown, in a mouse model, that gestational exposure to diesel exhaust results in increased cardiac hypertrophy, fibrosis and susceptibility to heart failure in the adult offspring following transverse aortic constriction. In this study, we have analyzed gene expression in neonatal cardiomyocytes after gestational exposure by RNA-sequencing and have identified 300 genes that are dysregulated, including many involved in cardiac metabolism.
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Overall design |
Female mice were paired with male mice for timed mating in FA. After observation of a vaginal plug, pregnant mice were put into FA or DE with exposures beginning at E0.5 and lasting until E17.5, at which point pregnant mice were transferred from the Northlake facility to the UW Medicine South Lake Union SPF vivarium for neonatal sample collection or weaned and kept until 11-12 weeks for adult sample collection. Upon birth, neonatal hearts were harvested, trimmed of surrounding vascular and atrial tissue, and dissociated as previously described. Dissociation was performed using 1mg/mL Liberase TH (Roche; Pleasanton, CA, USA) in 1X HBSS by incubating hearts at 37 degrees for 5 minutes in solution, with pipetting to release cells after incubation. Media containing released neonatal carciomyocytes (NCMs) was adjusted to 20% FBS-DMEM, and cellular dissociation was continued until the majority of cells were released. Cells were then filtered using a 70um sieve, re-eluted in 20% FBS-DMEM with 20uM Ara-C and incubated at 37 degrees for 1 hour to allow fibroblasts to attach onto the plate. After incubation, media with suspended cardiomyocytes was carefully removed, spun and purified cardiomyocyte pellets were collected and frozen at -80°C.
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Contributor(s) |
Goodson JM, MacDonald JW, Bammler TK, Chien W, Chin MT |
Citation(s) |
30975218 |
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Submission date |
Feb 16, 2018 |
Last update date |
Apr 17, 2019 |
Contact name |
James William MacDonald |
E-mail(s) |
[email protected]
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Organization name |
University of Washington
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Department |
Environmental and Occupational Health Sciences
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Street address |
4225 Roosevelt Way NE
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City |
Seattle |
State/province |
WA |
ZIP/Postal code |
98105-6099 |
Country |
USA |
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Platforms (1) |
GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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Samples (8)
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This SubSeries is part of SuperSeries: |
GSE110793 |
In utero exposure to diesel exhaust particulates is associated with alterations of the neonatal cardiomyocyte transcriptome, metabolic parturbation and a global decrease in DNA methylation. |
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Relations |
BioProject |
PRJNA434460 |
SRA |
SRP133012 |