A biomarker ORAI2 protein for the regulation of calcium signaling in neurons induced by microwave radiation
A microwave radiation, neuron technology, applied in the determination/inspection of microorganisms, biological testing, nervous system diseases, etc., can solve problems such as lack of biomarkers
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Embodiment 1
[0065] Example 1 Effect of Microwave Radiation on Calcium Ions in Neuron Cytoplasm, Mitochondria and Endoplasmic Reticulum
[0066] Fluo-4 AM, Mag-Fluo-AM and Rhod-2 were used to label calcium ions in the cytoplasm, mitochondria and endoplasmic reticulum of primary rat neurons, respectively. Self-designed real-time microwave radiation source, the radiation system is composed of signal generator, pulse power amplifier, circulator, bidirectional coupler, waveguide cavity and power meter for power monitoring, see figure 1 . The pulse-type power amplifier modulates and amplifies the microwave signal output by the signal source. The power module mainly supplies power for the power amplifier and the control monitoring board. The 30dB directional coupler monitors the coupled power signal in real time. transmission, preventing the reflected power signal from burning the power amplifier. The irradiation cavity adopts a pass-through waveguide structure, and the WR260 waveguide (inner ...
Embodiment 2
[0068] Example 2 Detection of neuron voltage-gated calcium channel current after microwave irradiation
[0069] Configure the inner and outer solution of the electrode, the inner solution is CsCl 140mM, TEA-Cl 10mM, Na2ATP 2mM, EGTA10mM, HEPES10mM, adjust the pH value to 7.3 with CsOH, filter with a microporous membrane (0.2μm); the outer solution is Choline-Cl 140mM, MgCl2 1mM, KCl 5mM, CaCl2 3mM, TEA-Cl 10mM, 4-AP 10mM, HEPES 10mM, Glucose 10mM, NaOH to adjust the pH value to 7.3, and filter through a microporous membrane (0.2μm). Hippocampal neurons were inoculated in 35mm culture dishes and cultured until the 7th day. After microwave radiation, the parameters of the patch clamp amplifier and data acquisition software were set. After the hippocampal neurons were sealed and membrane ruptured, the changes of VGCCs current were observed. The recording was performed immediately after irradiation, and the recording was completed within 6 hours. At least 15 cells were recorded in...
Embodiment 3
[0074] Example 3 Quantitative proteomics detection of rat hippocampus tissue after microwave irradiation
[0075] 80 secondary male Wistar rats (Academy of Military Medical Research) with a body weight of 220 ± 20 g were randomly divided into 4 groups (C, S, L and SL), and Group C was placed in a radiation box without radiation, and Group S , L and SL groups were exposed to 10mW / cm 2 The S-band, L-band and S- and L-band microwave composite radiations were irradiated for 6 minutes (after the radiation, the calcium signal in the rat hippocampal cells changed as similar to that in Examples 1 and 2). On the 7th day after normal feeding was resumed after radiation, hippocampus tissues (n=3 / group) of rats in groups C, S, L and SL were collected, total protein was extracted, and the protein was subjected to enzymatic hydrolysis. The peptide solution after enzymatic digestion was collected, labeled with iTRAQ, separated by reversed-phase chromatography under high pH conditions, and t...
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