Real-time detection of stimulus response in cultured neurons by high-intensity intermediate-frequency magnetic field exposure

Abstract

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.