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Detection and quantification of microRNA in cerebral microdialysate

BACKGROUND: Secondary brain injury accounts for a major part of the morbidity and mortality in patients with spontaneous aneurysmal subarachnoid hemorrhage (SAH), but the pathogenesis and pathophysiology remain controversial. MicroRNAs (miRNAs) are important posttranscriptional regulators of complem...

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Autores principales: Bache, Søren, Rasmussen, Rune, Rossing, Maria, Hammer, Niels Risør, Juhler, Marianne, Friis-Hansen, Lennart, Nielsen, Finn Cilius, Møller, Kirsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4438475/
https://www.ncbi.nlm.nih.gov/pubmed/25947950
http://dx.doi.org/10.1186/s12967-015-0505-1
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author Bache, Søren
Rasmussen, Rune
Rossing, Maria
Hammer, Niels Risør
Juhler, Marianne
Friis-Hansen, Lennart
Nielsen, Finn Cilius
Møller, Kirsten
author_facet Bache, Søren
Rasmussen, Rune
Rossing, Maria
Hammer, Niels Risør
Juhler, Marianne
Friis-Hansen, Lennart
Nielsen, Finn Cilius
Møller, Kirsten
author_sort Bache, Søren
collection PubMed
description BACKGROUND: Secondary brain injury accounts for a major part of the morbidity and mortality in patients with spontaneous aneurysmal subarachnoid hemorrhage (SAH), but the pathogenesis and pathophysiology remain controversial. MicroRNAs (miRNAs) are important posttranscriptional regulators of complementary mRNA targets and have been implicated in the pathophysiology of other types of acute brain injury. Cerebral microdialysis is a promising tool to investigate these mechanisms. We hypothesized that miRNAs would be present in human cerebral microdialysate. METHODS: RNA was extracted and miRNA profiles were established using high throughput real-time quantification PCR on the following material: 1) Microdialysate sampled in vitro from A) a solution of total RNA extracted from human brain, B) cerebrospinal fluid (CSF) from a neurologically healthy patient, and C) a patient with SAH; and 2) cerebral microdialysate and CSF sampled in vivo from two patients with SAH. MiRNAs were categorized according to their relative recovery (RR) and a pathway analysis was performed for miRNAs exhibiting a high RR in vivo. RESULTS: Seventy-one of the 160 miRNAs detected in CSF were also found in in vivo microdialysate from SAH patients. Furthermore specific miRNAs consistently exhibited either a high or low RR in both in vitro and in vivo microdialysate. Analysis of repeatability showed lower analytical variation in microdialysate than in CSF. CONCLUSIONS: MiRNAs are detectable in cerebral microdialysate; a large group of miRNAs consistently showed a high RR in cerebral microdialysate. Measurement of cerebral interstitial miRNA concentrations may aid in the investigation of secondary brain injury in neurocritical conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12967-015-0505-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-44384752015-05-21 Detection and quantification of microRNA in cerebral microdialysate Bache, Søren Rasmussen, Rune Rossing, Maria Hammer, Niels Risør Juhler, Marianne Friis-Hansen, Lennart Nielsen, Finn Cilius Møller, Kirsten J Transl Med Research BACKGROUND: Secondary brain injury accounts for a major part of the morbidity and mortality in patients with spontaneous aneurysmal subarachnoid hemorrhage (SAH), but the pathogenesis and pathophysiology remain controversial. MicroRNAs (miRNAs) are important posttranscriptional regulators of complementary mRNA targets and have been implicated in the pathophysiology of other types of acute brain injury. Cerebral microdialysis is a promising tool to investigate these mechanisms. We hypothesized that miRNAs would be present in human cerebral microdialysate. METHODS: RNA was extracted and miRNA profiles were established using high throughput real-time quantification PCR on the following material: 1) Microdialysate sampled in vitro from A) a solution of total RNA extracted from human brain, B) cerebrospinal fluid (CSF) from a neurologically healthy patient, and C) a patient with SAH; and 2) cerebral microdialysate and CSF sampled in vivo from two patients with SAH. MiRNAs were categorized according to their relative recovery (RR) and a pathway analysis was performed for miRNAs exhibiting a high RR in vivo. RESULTS: Seventy-one of the 160 miRNAs detected in CSF were also found in in vivo microdialysate from SAH patients. Furthermore specific miRNAs consistently exhibited either a high or low RR in both in vitro and in vivo microdialysate. Analysis of repeatability showed lower analytical variation in microdialysate than in CSF. CONCLUSIONS: MiRNAs are detectable in cerebral microdialysate; a large group of miRNAs consistently showed a high RR in cerebral microdialysate. Measurement of cerebral interstitial miRNA concentrations may aid in the investigation of secondary brain injury in neurocritical conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12967-015-0505-1) contains supplementary material, which is available to authorized users. BioMed Central 2015-05-07 /pmc/articles/PMC4438475/ /pubmed/25947950 http://dx.doi.org/10.1186/s12967-015-0505-1 Text en © Bache et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Bache, Søren
Rasmussen, Rune
Rossing, Maria
Hammer, Niels Risør
Juhler, Marianne
Friis-Hansen, Lennart
Nielsen, Finn Cilius
Møller, Kirsten
Detection and quantification of microRNA in cerebral microdialysate
title Detection and quantification of microRNA in cerebral microdialysate
title_full Detection and quantification of microRNA in cerebral microdialysate
title_fullStr Detection and quantification of microRNA in cerebral microdialysate
title_full_unstemmed Detection and quantification of microRNA in cerebral microdialysate
title_short Detection and quantification of microRNA in cerebral microdialysate
title_sort detection and quantification of microrna in cerebral microdialysate
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4438475/
https://www.ncbi.nlm.nih.gov/pubmed/25947950
http://dx.doi.org/10.1186/s12967-015-0505-1
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