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1.
Biosens Bioelectron ; 145: 111708, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31557560

RESUMO

Herein, we describe an electrophysiological based sensor that reproducibly monitors and quantifies in real-time collective migration and the formation of cell-cell junctions by C6 glioma cells seeded on top of electrodes. The signal amplitude and frequency generated by the migrating cells changed over time and these parameters were used to accurately calculate the migration speed. Electrophysiological measurements could also distinguish individual from collective cell migration. The migration of densely packed cells generated strong signals, while dispersed cells showed weak bioelectrical activity. We propose this electrophysiological technique as a cell-based biosensor to gain insight into the mechanisms of cooperative migration of cancer cells. Possible applications include screening for anti-migratory compounds, which may lead to the development of novel strategies for antineoplastic chemotherapy.


Assuntos
Técnicas Biossensoriais , Comunicação Celular/fisiologia , Movimento Celular/fisiologia , Glioma/fisiopatologia , Fenômenos Eletrofisiológicos , Glioma/diagnóstico , Humanos
2.
Front Neural Circuits ; 11: 80, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29109679

RESUMO

Astrocytes are neuroglial cells that exhibit functional electrical properties sensitive to neuronal activity and capable of modulating neurotransmission. Thus, electrophysiological recordings of astroglial activity are very attractive to study the dynamics of glial signaling. This contribution reports on the use of ultra-sensitive planar electrodes combined with low noise and low frequency amplifiers that enable the detection of extracellular signals produced by primary cultures of astrocytes isolated from mouse cerebral cortex. Recorded activity is characterized by spontaneous bursts comprised of discrete signals with pronounced changes on the signal rate and amplitude. Weak and sporadic signals become synchronized and evolve with time to higher amplitude signals with a quasi-periodic behavior, revealing a cooperative signaling process. The methodology presented herewith enables the study of ionic fluctuations of population of cells, complementing the single cells observation by calcium imaging as well as by patch-clamp techniques.


Assuntos
Astrócitos/fisiologia , Microeletrodos , Animais , Células Cultivadas , Córtex Cerebral/fisiologia , Fenômenos Eletrofisiológicos , Espaço Extracelular/fisiologia , Camundongos Endogâmicos C57BL
3.
Sci Rep ; 7(1): 14284, 2017 10 27.
Artigo em Inglês | MEDLINE | ID: mdl-29079771

RESUMO

Ultra-sensitive electrodes for extracellular recordings were fabricated and electrically characterized. A signal detection limit defined by a noise level of 0.3-0.4 µV for a bandwidth of 12.5 Hz was achieved. To obtain this high sensitivity, large area (4 mm2) electrodes were used. The electrode surface is also micro-structured with an array of gold mushroom-like shapes to further enhance the active area. In comparison with a flat gold surface, the micro-structured surface increases the capacitance of the electrode/electrolyte interface by 54%. The electrode low impedance and low noise enable the detection of weak and low frequency quasi-periodic signals produced by astrocytes populations that thus far had remained inaccessible using conventional extracellular electrodes. Signals with 5 µV in amplitude and lasting for 5-10 s were measured, with a peak-to-peak signal-to-noise ratio of 16. The electrodes and the methodology developed here can be used as an ultrasensitive electrophysiological tool to reveal the synchronization dynamics of ultra-slow ionic signalling between non-electrogenic cells.


Assuntos
Astrócitos/fisiologia , Potenciais da Membrana , Microeletrodos , Animais , Células Cultivadas , Córtex Cerebral/fisiologia , Capacitância Elétrica , Impedância Elétrica , Desenho de Equipamento , Compostos de Ouro , Camundongos Endogâmicos C57BL , Neurofisiologia/instrumentação , Cultura Primária de Células
4.
Sci Adv ; 2(12): e1600516, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28028533

RESUMO

Glioma patients often suffer from epileptic seizures because of the tumor's impact on the brain physiology. Using the rat glioma cell line C6 as a model system, we performed long-term live recordings of the electrical activity of glioma populations in an ultrasensitive detection method. The transducer exploits large-area electrodes that maximize double-layer capacitance, thus increasing the sensitivity. This strategy allowed us to record glioma electrical activity. We show that although glioma cells are nonelectrogenic, they display a remarkable electrical burst activity in time. The low-frequency current noise after cell adhesion is dominated by the flow of Na+ ions through voltage-gated ion channels. However, after an incubation period of many hours, the current noise markedly increased. This electric bursting phenomenon was not associated with apoptosis because the cells were viable and proliferative during the period of increased electric activity. We detected a rapid cell culture medium acidification accompanying this event. By using specific inhibitors, we showed that the electrical bursting activity was prompted by extracellular pH changes, which enhanced Na+ ion flux through the psalmotoxin 1-sensitive acid-sensing ion channels. Our model of pH-triggered bursting was unambiguously supported by deliberate, external acidification of the cell culture medium. This unexpected, acidosis-driven electrical activity is likely to directly perturb, in vivo, the functionality of the healthy neuronal network in the vicinity of the tumor bulk and may contribute to seizures in glioma patients.


Assuntos
Fenômenos Eletrofisiológicos , Glioma/fisiopatologia , Concentração de Íons de Hidrogênio , Neurônios/citologia , Animais , Linhagem Celular Tumoral , Humanos , Rede Nervosa , Ratos , Canais de Sódio/fisiologia
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