High-intensity and low frequency (1–100 kHz) time-varying electromagnetic fields stimulate the human body through excitation of the nervous system. In power frequency range (50/60 Hz), a frequency-dependent threshold of the external electric field-induced neuronal modulation in cultured neuronal networks was used as one of the biological indicator in international guidelines; however, the threshold of the magnetic field-induced neuronal modulation has not been elucidated. In this study, we exposed rat brain-derived neuronal networks to a high-intensity power frequency magnetic field (hPF-MF), and evaluated the modulation of synchronized bursting activity using a multi-electrode array (MEA)-based extracellular recording technique. As a result of short-term hPF-MF exposure (50–400 mT root-mean-square (rms), 50 Hz, sinusoidal wave, 6 s), the synchronized bursting activity was increased in the 400 mT-exposed group. On the other hand, no change was observed in the 50–200 mT-exposed groups. In order to clarify the mechanisms of the 400 mT hPF-MF exposure-induced neuronal response, we evaluated it after blocking inhibitory synapses using bicuculline methiodide (BMI); subsequently, increase in bursting activity was observed with BMI application, and the response of 400 mT hPF-MF exposure disappeared. Therefore, it was suggested that the response of hPF-MF exposure was involved in the inhibitory input. Next, we screened the inhibitory pacemaker-like neuronal activity which showed autonomous 4–10 Hz firing with CNQX and D-AP5 application, and it was confirmed that the activity was reduced after 400 mT hPF-MF exposure. Comparison of these experimental results with estimated values of the induced electric field (E-field) in the culture medium revealed that the change in synchronized bursting activity occurred over 0.3 V/m, which was equivalent to the findings of a previous study that used the external electric fields. In addition, the results suggested that the potentiation of neuronal activity after 400 mT hPF-MF exposure was related to the depression of autonomous activity of pacemaker-like neurons. Our results indicated that the synchronized bursting activity was increased by hPF-MF exposure (E-field: >0.3 V/m), and the response was due to reduced inhibitory pacemaker-like neuronal activity.
Threshold values of neuronal stimulation and modulation associated with exposure to time-varying electromagnetic fields contribute to establishing human protection guidelines and standards. However, biological evidence of threshold values in the intermediate-frequency range is limited. Additionally, although it is known that dendrites, a type of unmyelinated neuronal fibre, play an important role in information processing in the central nervous system, the stimulus threshold in dendrites has not been sufficiently investigated. We evaluated the excitation site-specific stimulus response of rat brain-derived cultured neurons by using a 20 kHz high-intensity intermediate-frequency magnetic field (hIF-MF) exposure system, a non-conductive fibre-optic imaging (NCFI) system, combined with a micro-patterning technique. Our hIF-MF exposure and NCFI system permitted real-time detection of the intracellular calcium ([Ca2+]i) spikes in neuronal cell bodies or unmyelinated neuronal fibres during exposure to a 20 kHz, 70 mT (peak), burst-type sinusoidal wave hIF-MF. Dosimetry of the induced electric fields intensities in the extracellular solution indicated that about 50% of unmyelinated neuronal fibres respond at about 147 V m-1. In contrast, the threshold of the [Ca2+]i spikes in neuronal cell bodies were lower than that in unmyelinated neuronal fibres. Our results provide a basis for understanding site-specific differences in the responses of cultured neurons to hIF-MFs.
International Commission on Non-Ionizing Radiation Protection (ICNIRP) guidelines describes addition equation for electrical stimulation, relevant for frequencies up to 10 MHz, with respect to simultaneous exposure to magnetic fields with multiple frequencies. However, there is no literature which estimated the threshold of electromagnetic field exposure of multiple frequencies. Therefore, the purpose of this article is to estimate the threshold of nerve excitation caused by the application of current with multiple frequencies. In this article, combinations of two kinds of frequencies f 1 and f 2 were investigated for the applying current. The Frankenhaeuser-Huxley model was employed to analyze nerve excitation effects. The ratios of the applied current value to the threshold current value were kept at the same value for each frequency f 1
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