Investigation On Modulation Of Cell Signaling Pathway And Mechanism By Femtosecond Laser

Cell signaling transduction is fundamental to regulation of cellular physiological processes.Artificial modulation of cellular signal molecule is the key technology in numerous life science and clinical medicine studies.Chemical drug stimulation is the most used treatment means in modulating cell signaling to regulate cellular processes.In recent years,with the development of laser technology,laser stimulation has gradually become a novel non-invasive,low toxicity and more precise and controllable method which has attracted more and more attention in modulation of cell signal transduction.In this regards,we combined femtosecond laser and confocal microscopy to develop an accurate femtosecond laser stimulation method on cells or organelles at a high temporal and spatial resolution level.It is found that femtosecond laser stimulation could elicit many molecular signal changes in cells.It was of great significance in research of cell function regulation.In this paper,based on the researches in activation of calcium ion channels by femtosecond laser,it was found that femtosecond laser activated some small molecular signals such as calcium,reactive oxygen species(ROS)and mitochondrial membrane potential.The independence of activation of calcium and ROS by femtosecond laser was also demonstrated.All these approaches provided experimental basis for further activation of cell signaling pathways and organelle signals.It was found that femtosecond laser stimulation could induce the migrating into nucleus of phosphorylated extracellular regulated kinases 2(ERK2).The activation of ERK2 signaling pathway was verified.The molecular mechanism of activation of ERK2 signaling pathway by femtosecond laser was further clarified through more related experiments.In this study,femtosecond laser stimulation could cause morphology changes of single mitochondrion.The types of morphology changes were influenced by the intensity of femtosecond laser stimulation.According to this approach,femtosecond laser treatment could activate mitochondrial signals such as membrane potential change,opening of mitochondrial permeability transition pore(mPTP)and mitoflash activities.By quantitative analysis the characteristics of photo-induced mitoflash,the technique of photo-activated mitoflash by femtosecond laser was quantified and standardized.In addition,based on the investigation of two photon optogenetics,this paper proposed a low power density irradiation method by femtosecond laser,which could activate CRY2/CIB1 optogenetic tool by multi-photon absorption.In this study,by using CRY2/CIB1 optogenetic tool,regulation of cell apoptosis was successfully realized.The range of femtosecond laser wavelength which could activate CRY2 was also investigated.In this paper,a series of work explored the ability of femtosecond laser to modulate multiple cell signals including small signal molecules,activation of protein kinase signaling pathway,organelles and exogenous protein signals.These studies provided a novel method and important theory for optical regulating cell functions by femtosecond laser.