AAV-Syn-BDNF-EGFP Virus Construct Exerts Neuroprotective Action on the Hippocampal Neural Network during Hypoxia In Vitro

Autor: Ekaterina A. Epifanova, Maria Vedunova, Alexey A. Babaev, Roman S. Yarkov, Elena V. Mitroshina, Maria S. Gavrish, Alexandra V. Usenko, Tatiana A. Mishchenko
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
multielectrode arrays
Green Fluorescent Proteins
Hippocampal formation
Biology
Neuroprotection
Hippocampus
Catalysis
Article
Viral vector
Inorganic Chemistry
lcsh:Chemistry
03 medical and health sciences
Mice
adeno-associated virus vector
0302 clinical medicine
Calcium imaging
primary hippocampal cell cultures
Neurotrophic factors
medicine
Animals
Viability assay
Physical and Theoretical Chemistry
Hypoxia
Hypoxia
Brain
Molecular Biology
lcsh:QH301-705.5
Spectroscopy
Cells
Cultured
Brain-derived neurotrophic factor
Organic Chemistry
brain-derived neurotrophic factor
Cerebral hypoxia
General Medicine
Dependovirus
medicine.disease
cerebral hypoxia
Computer Science Applications
Cell biology
Mice
Inbred C57BL
calcium imaging
030104 developmental biology
BDNF
lcsh:Biology (General)
lcsh:QD1-999
neuroprotection
030217 neurology & neurosurgery
Zdroj: International Journal of Molecular Sciences, Vol 19, Iss 8, p 2295 (2018)
International Journal of Molecular Sciences
Volume 19
Issue 8
ISSN: 1422-0067
Popis: Brain-derived neurotrophic factor (BDNF) is one of the key signaling molecules that supports the viability of neural cells in various brain pathologies, and can be considered a potential therapeutic agent. However, several methodological difficulties, such as overcoming the blood&ndash
brain barrier and the short half-life period, challenge the potential use of BDNF in clinical practice. Gene therapy could overcome these limitations. Investigating the influence of viral vectors on the neural network level is of particular interest because viral overexpression affects different aspects of cell metabolism and interactions between neurons. The present work aimed to investigate the influence of the adeno-associated virus (AAV)-Syn-BDNF-EGFP virus construct on neural network activity parameters in an acute hypobaric hypoxia model in vitro. Materials and methods. An adeno-associated virus vector carrying the BDNF gene was constructed using the following plasmids: AAV-Syn-EGFP, pDP5, DJvector, and pHelper. The developed virus vector was then tested on primary hippocampal cultures obtained from C57BL/6 mouse embryos (E18). Acute hypobaric hypoxia was induced on day 21 in vitro. Spontaneous bioelectrical and calcium activity of neural networks in primary cultures and viability tests were analysed during normoxia and during the posthypoxic period. Results. BDNF overexpression by AAV-Syn-BDNF-EGFP does not affect cell viability or the main parameters of spontaneous bioelectrical activity in normoxia. Application of the developed virus construct partially eliminates the negative hypoxic consequences by preserving cell viability and maintaining spontaneous bioelectrical activity in the cultures. Moreover, the internal functional structure, including the activation pattern of network bursts, the number of hubs, and the number of connections within network elements, is also partially preserved. BDNF overexpression prevents a decrease in the number of cells exhibiting calcium activity and maintains the frequency of calcium oscillations. Conclusion. This study revealed the pronounced antihypoxic and neuroprotective effects of AAV-Syn-BDNF-EGFP virus transduction in an acute normobaric hypoxia model.
Databáze: OpenAIRE
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