title: DNA Content in Extracellular Vesicles Isolated from Porcine Coronary Venous Blood Directly after Myocardial Ischemic Preconditioning.
abstract: 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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Anesthesiology and Intensive Care Medicine, Institute of Clinical Science, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Kristina
authors.identifier: http://orcid.org/0000-0002-2173-2978
authors.initials: K
authors.lastname: Svennerholm
authors.affiliation: Anesthesiology and Intensive Care Medicine, Institute of Surgical and Perioperative Science, Umeå University, Umeå, Sweden.
authors.forename: Pouria
authors.initials: P
authors.lastname: Rodsand
authors.affiliation: Cardiology, Institute of Heart Centre and Department of Public Health and Clinical Medicine, Medicine, Umeå University, Umeå, Sweden.
authors.forename: Urban
authors.initials: U
authors.lastname: Hellman
authors.affiliation: Cardiology, Institute of Heart Centre and Department of Public Health and Clinical Medicine, Medicine, Umeå University, Umeå, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Waldenström
authors.affiliation: Pathology, Institute of Medical Biosciences, Umeå University, Umeå, Sweden.
authors.forename: Marie
authors.initials: M
authors.lastname: Lundholm
authors.affiliation: National Bioinformatics Infrastructure Sweden (NBIS), Institute of Biology, Lund University, Lund, Sweden.
authors.forename: Dag
authors.initials: D
authors.lastname: Ahrén
authors.affiliation: Anesthesiology and Intensive Care Medicine, Institute of Clinical Science, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Biber
authors.affiliation: Clinical Chemistry, Institutet of Medical Sciences, Uppsala University, Uppsala, Sweden.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Ronquist
authors.affiliation: Anesthesiology and Intensive Care Medicine, Institute of Surgical and Perioperative Science, Umeå University, Umeå, Sweden.
authors.forename: Michael
authors.initials: M
authors.lastname: Haney
mesh_terms: D000818:Animals; D003331:Coronary Vessels / Q000378:metabolism* / Q000473:pathology; D004247:DNA / Q000097:blood; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D019157:Ischemic Preconditioning, Myocardial / Q000379:methods*; D009203:Myocardial Infarction / Q000097:blood / Q000473:pathology; D015428:Myocardial Reperfusion Injury / Q000097:blood / Q000503:physiopathology*; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D013552:Swine
chemical_list: D004247:DNA