- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Src Kinase Controls Signaling Pathways In Sensory Neuron Triggered By Low-Power Infrared Radiation
摘要： Low-power (non-thermal) infrared (IR) radiation with the wavelength of 10.6 μm activates the Na,K-ATPase transducer function in sensory neurons, which is manifested in decrease of NaV1.8 channel voltage sensitivity at the cellular membrane level and in inhibition of growth of chick embryo dorsal root ganglia neurites at the tissue level. It is shown that the effect of low-power IR radiation is totally blocked by a specific Src kinase inhibitor PP2. Upon irradiation on the background of PP2, the effective charge of NaV1.8 channel activation gating system does not differ from its control value in patch-clamp experiments, and the area index of sensory ganglia neurites growth remains unchanged as compared to the control in organotypic tissue culture. The data obtained demonstrate that Src kinase is involved in intracellular signaling pathways triggered by CO2 laser low-power IR radiation by the transducer-activated mechanism. This is the first indication that in primary sensory neuron the signals of low-power IR radiation are sensed, amplified and transduced by the Na,K-ATPase/Src complex and not by G proteins.
关键词： Src kinase,infrared radiation,Na,K-ATPase,organotypic nerve tissue culture,NaV1.8 channels,sensory neuron,patch-clamp method
更新于2025-11-21 11:24:58
-
Quick visualization of neurons in brain tissues using an optical clearing technique
摘要： Neurons are classified into several morphological types according to the locations of their somata and the branching patterns of their axons and dendrites. Recent studies suggest that these morphological features are related to their physiological properties, including firing characteristics, responses to neuromodulators, and wiring patterns. Therefore, rapid morphological identification of electrophysiologically recorded neurons promises to advance our understanding of neuronal circuits. One of the most common anatomical cell identification methods is neuronal reconstruction with biocytin delivered through whole-cell patch-clamp pipettes. However, conventional reconstruction methods usually take longer than 24 h and limit the throughput of electrophysiological experiments. Here, we developed a quick, simple cell reconstruction method by optimizing the tissue clearing protocol ScaleSQ. We found that adding 200 mM NaCl almost entirely prevented tissue swelling without compromising optical clearing ability. This solution, termed IsoScaleSQ, allowed us to increase the transparency of the gray matter of 500-μm-thick slices within 30 min, meaning that the total time required to reconstruct whole-cell recorded neurons was reduced to 1 h. This novel method will improve the efficacy and effectiveness of electrophysiological experiments linked to cell morphology.
关键词： Optical clearing,Isotropic,Visualization,Patch-clamp recording,ScaleSQ
更新于2025-09-23 15:23:52
-
Fluorescence-based high throughput screening technologies for natural chloride ion channel blockers
摘要： Chloride channels represent a group of potential drug targets, their blockers showed significant protecting effect on impaired cells by modulating apoptosis, autophagy and other cell signals. However, clinical drugs with chloride channel inhibitory properties have not yet been developed. Natural product extract becomes an underlying candidate satisfied the clinical requirements for its low toxicity, low cost and abundant sources. Here, a fluorescence-based EYFP-H148Q/I153L-HeLa cell line model was constructed by molecular cloning and verified by Real-time PCR and Western Blotting assay. By using this chloride channel blocker screening model, 7 hit compounds chosen from 6988 natural compounds showed the channel blocking activity. Then the hit compounds were further validated by electrophysiological patch-clamp analysis. Our study preliminarily identified PC-4 as a new chloride channel inhibitors and demonstrated the reliability and sensitivity of fluorescence-based high throughput screening technologies for discovery of biologically active compounds from natural herbal compounds.
关键词： natural compound,high throughput screening,chloride channel blocker,patch clamp
更新于2025-09-23 15:21:21