Analysis of extracellular RNA in cerebrospinal fluid
| Autor: | Jorge I. Arango, Johnny C. Akers, Joseph F. Quinn, Fred H. Hochberg, Theresa A. Lusardi, Bob S. Carter, Christina A. Harrington, Jay Ian Phillips, Erika Duggan, Douglas Galasko, Ashish Yeri, Trevor J. McFarland, John P. Nolan, Karen Messer, Rebecca Reiman, Babett Lind, M. Yashar S. Kalani, Julie A. Saugstad, Amanda Courtright, Kendall Van Keuren-Jensen, P. David Adelson |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Histology Biology Neurodegenerative Article cerebrospinal fluid Flow cytometry 03 medical and health sciences Cerebrospinal fluid Rare Diseases Glioma microRNA medicine Genetics Acquired Cognitive Impairment 2.1 Biological and endogenous factors lcsh:QH573-671 Aetiology Cancer medicine.diagnostic_test lcsh:Cytology Neurosciences RNA Cell Biology Extracellular vesicle Extracellular vesicles medicine.disease Molecular biology extracellular RNA 3. Good health Brain Disorders Brain Cancer 030104 developmental biology Neurological Dementia RNA extraction Biochemistry and Cell Biology Transferred Article neurological diseases Extracellular RNA Biotechnology |
| Zdroj: | Journal of Extracellular Vesicles Journal of extracellular vesicles, vol 6, iss 1 Journal of Extracellular Vesicles, Vol 6, Iss 1 (2017) |
| ISSN: | 2001-3078 |
| Popis: | We examined the extracellular vesicle (EV) and RNA composition of pooled normal cerebrospinal fluid (CSF) samples and CSF from five major neurological disorders: Alzheimer’s disease (AD), Parkinson’s disease (PD), low-grade glioma (LGG), glioblastoma multiforme (GBM), and subarachnoid haemorrhage (SAH), representing neurodegenerative disease, cancer, and severe acute brain injury. We evaluated: (I) size and quantity of EVs by nanoparticle tracking analysis (NTA) and vesicle flow cytometry (VFC), (II) RNA yield and purity using four RNA isolation kits, (III) replication of RNA yields within and between laboratories, and (IV) composition of total and EV RNAs by reverse transcription–quantitative polymerase chain reaction (RT-qPCR) and RNA sequencing (RNASeq). The CSF contained ~106 EVs/μL by NTA and VFC. Brain tumour and SAH CSF contained more EVs and RNA relative to normal, AD, and PD. RT-qPCR and RNASeq identified disease-related populations of microRNAs and messenger RNAs (mRNAs) relative to normal CSF, in both total and EV fractions. This work presents relevant measures selected to inform the design of subsequent replicative CSF studies. The range of neurological diseases highlights variations in total and EV RNA content due to disease or collection site, revealing critical considerations guiding the selection of appropriate approaches and controls for CSF studies. |
| Databáze: | OpenAIRE |
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