Mouse Paneth Cell-Enriched Proteome Enabled by Laser Capture Microdissection

被引:3
作者
Woo, Jongmin [1 ]
Schoenfeld, Madeline [1 ]
Sun, Xinguo [1 ]
Iraguha, Thierry [1 ]
Zhou, Zhanxiang [1 ,2 ]
Zhang, Qibin [1 ,3 ]
机构
[1] Univ North Carolina Greensboro, Ctr Translat Biomed Res, North Carolina Res Campus, Kannapolis, NC 28081 USA
[2] Univ North Carolina Greensboro, Dept Nutr, Greensboro, NC 27402 USA
[3] Univ North Carolina Greensboro, Dept Chem & Biochem, Greensboro, NC 27402 USA
基金
美国国家卫生研究院;
关键词
Paneth cell; proteomics; laser capture microdissection; toluidine blue; mouse small intestine; villi; LYSOZYME;
D O I
10.1021/acs.jproteome.2c00311
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Paneth cells are antimicrobial peptide-secreting cells located at the base of the crypts of the small intestine. The proteome of Paneth cells is not well defined because of their coexistence with stem cells, making it difficult to culture Paneth cells alone in vitro. Using a simplified toluidine blue O method for staining mouse intestinal tissue, laser capture microdissection (LCM) to isolate cells from the crypt region, and surfactant-assisted one-pot protein digestion, we identified more than 1300 proteins from crypts equivalent to 18,000 cells. Compared with the proteomes of villi and smooth muscle regions, the crypt proteome is highly enriched in defensins, lysozymes, and other antimicrobial peptides that are characteristic of Paneth cells. The sensitivity of the LCM-based proteomics approach was also assessed using a smaller number of cell equivalent tissues: a comparable proteomic coverage can be achieved with 3600 cells. This work is the first proteomics study of intestinal tissue enriched with Paneth cells. The simplified workflow enables profiling of Paneth cell-associated pathological changes at the proteome level directly from frozen intestinal tissue. It may also be useful for proteomics studies of other spatially resolved cell types from other tissues.
引用
收藏
页码:2435 / 2442
页数:8
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