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DNA Content in Extracellular Vesicles Isolated from Porcine Coronary Venous Blood Directly after Myocardial Ischemic Preconditioning

Artikel i vetenskaplig tidskrift
Författare Kristina Svennerholm
P. Rodsand
U. Hellman
A. Waldenstrom
M. Lundholm
D. Ahren
Björn Biber
G. Ronquist
M. Haney
Publicerad i Plos One
Volym 11
Nummer/häfte 7
Sidor e0159105
ISSN 1932-6203
Publiceringsår 2016
Publicerad vid Institutionen för kliniska vetenskaper, Avdelningen för anestesiologi och intensivvård
Sidor e0159105
Språk en
Länkar dx.doi.org/10.1371/journal.pone.015...
Ämnesord mediated transfer, human prostasomes, genomic dna, cells, exosomes, infarction, mechanism, cancer, heart, Science & Technology - Other Topics
Ämneskategorier Klinisk medicin

Sammanfattning

Background Extracellular vesicles (EV) are nano-sized membranous structures released from most cells. They have the capacity to carry bioactive molecules and gene expression signals between cells, thus mediating intercellular communication. It is believed that EV confer protection after ischemic preconditioning (IPC). We hypothesize that myocardial ischemic preconditioning will lead to rapid alteration of EV DNA content in EV collected from coronary venous effluent. In a porcine myocardial ischemic preconditioning model, EV were isolated from coronary venous blood before and after IPC by differential centrifugation steps culminating in preparative ultracentrifugation combined with density gradient ultracentrifugation. The EV preparation was validated, the DNA was extracted and further characterized by DNA sequencing followed by bioinformatics analysis. Porcine genomic DNA fragments representing each chromosome, including mitochondrial DNA sequences, were detected in EV isolated before and after IPC. There was no difference detected in the number of sequenced gene fragments (reads) or in the genomic coverage of the sequenced DNA fragments in EV isolated before and after IPC. Gene ontology analysis showed an enrichment of genes coding for ion channels, enzymes and proteins for basal metabolism and vesicle biogenesis and specific cardiac proteins. This study demonstrates that porcine EV isolated from coronary venous blood plasma contain fragments of DNA from the entire genome, including the mitochondria. In this model we did not find specific qualitative or quantitative changes of the DNA content in EV collected immediately after an in vivo myocardial IPC provocation. This does not rule out the possibility that EV DNA content changes in response to myocardial IPC which could occur in a later time frame.

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