视网膜感光细胞对仿生电场刺激的非矢量应答  

Nonvectorial responses in photoreceptor cells stimulated by electrical fields

作　　者：杨极 王青锋 林蒙 何凯文 雷新兰 高美玲 潘登 金子兵 YANG Ji;WANG QingFeng;LIN Meng;HE KaiWen;LEI XinLan;GAO MeiLing;PAN Deng;JIN ZiBing(Institute of Stem Cell Research;Division of Ophthalmic Genetics, The Eye Hospital, Laboratory for Stem Cell ＆ Retinal Regeneration, Wenzhou Medical University, Wenzhou 325027, China;2 State Key Laboratory of Ophthalmology, Optometry and Vision Science, Wenzhou 325027, China)

机构地区：[1]温州医科大学干细胞研究所,温州医科大学附属眼视光医院视网膜再生医疗、眼科遗传学研究实验室,温州325027 [2]视光学与视觉科国家重点实验室,温州325027

出　　处：《中国科学：生命科学》2018年第5期544-555,共12页Scientia Sinica(Vitae)

基　　金：国家自然科学基金(批准号：81371059,81522014,81600772); 国家重点研发计划(批准号：2017YFA0105300,2017YFB0403702,2013CB967502); 浙江省自然科学基金(批准号：LQ17H120005); 浙江省重点研发计划(批准号：2015C03029); 温州科技创新团队(批准号：C20150004); 浙江省新苗人才计划(批准号：2016R413027)资助

摘　　要：正常状态下眼组织的细胞暴露于生物体内生电场中,故电场可以调控眼组织细胞一系列的生物学性能.细胞对电场信号的应答可分为两个方面,细胞对电场矢量的应答(迁移、定向生长等)和细胞对电场刺激的非矢量应答(增殖、凋亡等),而细胞对电场矢量的应答是最常见的应答现象.同时,细胞外的电场也能引起细胞内的信号转导,进而调控细胞的行为,但是对细胞内的应答机制目前知之甚少,所以对细胞非矢量性应答的机制也了解甚少.而感光细胞对电场刺激非矢量性应答的分子机制目前研究处于空白阶段,故本研究着重于探究感光细胞对于电场刺激的非矢量性应答的机制.为了探究电场刺激对感光细胞的影响,本课题组自主研发了一款电场刺激仪,研究发现,60和90 m V/mm强度的电场刺激能提高细胞活性,促进细胞增殖.检测基因表达谱,利用通路分析发现,Ca^(2+)离子依赖的ERK通路在细胞对电场刺激的应答中扮演重要角色.实验结果进一步证明,电场刺激促进Ca^(2+)离子内流进而改变了感光细胞的生长平衡,可以预见电场影响感光细胞的离子转运在感光细胞对电场刺激的应答过程中具有重要的意义.总而言之,本课题组发现,电场影响细胞内外的Ca^(2+)离子流,引发细胞内一系列的级连反应,使细胞的生长状态发生了改变.The eye possesses various different types of electrical properties, and ocular cells are exposed to endogenous electrical fields（EFs）under natural conditions. Cellular behaviors that are regulated by EFs include vectorial（migration, orientation, and elongation） and nonvectorial responses（apoptosis and proliferation）. The aim of this study was to investigate the nonvectorial responses of photoreceptor cells stimulated by EFs and to decipher the underlying molecular mechanisms. To investigate the effects of endogenous EFs on the cell behavior, we designed a customized electrical instrument that was mimetic of endogenous electrical activity. We observed that 60 and 90 m V/mm voltage gradients dramatically enhanced cell proliferation. Gene expression profiling, together with signaling pathway and gene ontology analyses, collectively demonstrated that the Ca^2＋-dependent extracellular signal-regulated kinase pathway is involved in cellular responses to EFs. Moreover, the results of our study clearly demonstrated that EFs promote photoreceptor cell proliferation via increased Ca^2＋ influx, which subsequently activates the ERK pathway. EF stimulation affects ionic fluxes in photoreceptor cells and is an important regulator of some important aspects of photoreceptor cell behavior. In conclusion, the findings of our study offer new insights on how EFs affect intracellular Ca^2＋, which further influences intracellular signal transduction and cell behavior.

关 键 词：电场 细胞行为 ERK通路 Ca2+离子流 

分 类 号：Q274[生物学—细胞生物学]