| Autor: |
Jahnke HG; Centre for Biotechnology and Biomedicine, Biochemical Cell Technology, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany., Te Kamp V; Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald, Germany., Prönnecke C; Centre for Biotechnology and Biomedicine, Biochemical Cell Technology, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany., Schmidt S; Centre for Biotechnology and Biomedicine, Biochemical Cell Technology, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany., Azendorf R; Centre for Biotechnology and Biomedicine, Biochemical Cell Technology, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany., Klupp B; Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald, Germany., Robitzki AA; Division Management for Biology, Chemistry and Process Engineering, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, 76344 Eggenstein-Leopoldshafen, Germany., Finke S; Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald, Germany. |
| Abstrakt: |
Development and optimisation of bioelectronic monitoring techniques like microelectrode array-based field potential measurement and impedance spectroscopy for the functional, label-free and non-invasive monitoring of in vitro neuronal networks is widely investigated in the field of biosensors. Thus, these techniques were individually used to demonstrate the capabilities of, e.g., detecting compound-induced toxicity in neuronal culture models. In contrast, extended application for investigating the effects of central nervous system infecting viruses are rarely described. In this context, we wanted to analyse the effect of herpesviruses on functional neuronal networks. Therefore, we developed a unique hybrid bioelectronic monitoring platform that allows for performing field potential monitoring and impedance spectroscopy on the same microelectrode. In the first step, a neuronal culture model based on primary hippocampal cells from neonatal rats was established with reproducible and stable synchronised electrophysiological network activity after 21 days of cultivation on microelectrode arrays. For a proof of concept, the pseudorabies model virus PrV Kaplan-ΔgG-GFP was applied and the effect on the neuronal networks was monitored by impedance spectroscopy and field potential measurement for 72 h in a multiparametric mode. Analysis of several bioelectronic parameters revealed a virus concentration-dependent degeneration of the neuronal network within 24-48 h, with a significant early change in electrophysiological activity, subsequently leading to a loss of activity and network synchronicity. In conclusion, we successfully developed a microelectrode array-based hybrid bioelectronic measurement platform for quantitative monitoring of pathologic effects of a herpesvirus on electrophysiological active neuronal networks. |
