김강훈 (연세대학교, 현 Monell Chemical Sense Center (University of Pennsylvania) ) | 한빛사논문 > 한빛사

한빛사논문

조회604

연세대학교, 현 Monell Chemical Sense Center (University of Pennsylvania)

CV File 280 kb

ACS Nano, Publication Date:May 19, 2023 | https://doi.org/10.1021/acsnano.2c10664 Molecular Decrowding by Tissue Expansion Allows Precise Determination of the Spatial Distribution of Synaptic Proteins at a Nanometer Scale by exTEM

Kang-Hoon Kim^1,2,7, Jubeen Yoon^1,2,7, Christian P. Macks^1,2,7, Han-Eol Park^1,2,3, Jihyun Youn^1,2,4, Jung-uk Lee^1,2, Minji An^1,2, Jongseong Park^1,2, Jaewon Ko^5,6, and Chang Ho Sohn^1,2*

¹Center for Nanomedicine, Institute for Basic Science (IBS), Yonsei University, Seoul 03722, Republic of Korea
²Graduate Program of Nano Biomedical Engineering (NanoBME), Advanced Science Institute, Yonsei University, Seoul 03722, Republic of Korea
³School of Biological Sciences, Seoul National University, Seoul 08826, Republic of Korea
⁴Department of Life Science and Biotechnology, Underwood International College, Yonsei University, Seoul 03722, Republic of Korea
⁵Department of Brain Sciences, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 42988, Republic of Korea
⁶Center for Synapse Diversity and Specificity, DGIST, Daegu 42988, Republic of Korea
⁷These authors contributed equally to this work.

^*Corresponding author: Chang Ho Sohn

Abstract

To understand how the molecular machinery of synapses works, it is essential to determine an inventory of synaptic proteins at a subsynaptic resolution. Nevertheless, synaptic proteins are difficult to localize because of the low expression levels and limited access to immunostaining epitopes. Here, we report on the exTEM (epitope-exposed by expansion-transmission electron microscopy) method that enables the imaging of synaptic proteins in situ. This method combines TEM with nanoscale resolution and expandable tissue-hydrogel hybrids for enhanced immunolabeling with better epitope accessibility via molecular decrowding, allowing successful probing of the distribution of various synapse-organizing proteins. We propose that exTEM can be employed for studying the mechanisms underlying the regulation of synaptic architecture and function by providing nanoscale molecular distribution of synaptic proteins in situ. We also envision that exTEM is widely applicable for investigating protein nanostructures located in densely packed environments by immunostaining of commercially available antibodies at nanometer resolution.

논문정보

형식Research article
게재일2023년 05월 (BRIC 등록일 2023-05-22)
연구진국내 연구진
분야 의약학 > 유전 및 유전체의학

댓글 작성 로그인

CV 열람하기

연락처 보기