利用果蝇模型筛选可用于动物组织光电关联的绿色荧光蛋白变体  

Screening green fluorescent protein variants for animal tissue correlative light and electron microscopy using Drosophila melanogaster

作　　者：陈捷 石丹妮 齐佳苗 孙玲 杨宇丰 王道彪 CHEN Jie;SHI Danni;QI Jiamiao;SUN Ling;YANG Yufeng;WANG Daobiao(Institute of Life Sciences,Fuzhou University,Fuzhou Fujian 350116,China)

机构地区：[1]福州大学生命科学研究所,福建福州350116

出　　处：《电子显微学报》2025年第1期81-90,共10页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金资助项目(No.32170658).

摘　　要：荧光蛋白能够将分子分布与超微结构联系起来,对于光电关联(CLEM)技术的应用至关重要。然而,常规的绿色荧光蛋白及其变体在活体组织的电子显微镜(EM)制样过程中,常因诸多不可控因素导致荧光信号显著减弱,这限制了它们作为光电关联分子探针的应用。本研究在果蝇大脑神经束膜胶质细胞中特异性表达多种GFP变体(mEmerald、hfYFP、mhYFP和CoGFPv0),以评估它们在活体组织光电关联中的应用潜力。通过提高表达量和靶向线粒体等策略,本研究最终筛选出适用于CLEM技术在果蝇神经系统研究中的CoGFPv0。本工作表明,CoGFPv0可作为一种具有良好应用前景的光电关联分子探针,特别适用于复杂的活体组织环境。Fluorescent proteins are crucial for correlating molecular distribution with ultrastructural details,which is essential for applications in correlative light and electron microscopy(CLEM).However,conventional green fluorescent proteins and their variants often experience significant fluorescence signal loss during electron microscopy(EM)sample preparation in live tissues,due to various uncontrollable factors,limiting their use as molecular probes for CLEM.In this study,multiple GFP variants(mEmerald,hfYFP,mhYFP,and CoGFPv0)were specifically expressed in the neuropil glial cells of the Drosophila brain to evaluate their potential for CLEM applications in vivo.Through strategies such as increasing expression levels and targeting mitochondria,this study ultimately identified CoGFPv0 as a suitable probe for CLEM in Drosophila nervous system research.This work demonstrates that CoGFPv0 shows strong potential as an effective molecular probe for CLEM,especially in complex live tissue environments.

关 键 词：光电关联技术 荧光蛋白 GFP变体 CoGFPv0 果蝇 

分 类 号：Q78[生物学—分子生物学] Q963[理学—分析化学] O657[理学—化学] Q336