pr2c00311_si_003.xlsx (34.02 kB)
Mouse Paneth Cell-Enriched Proteome Enabled by Laser Capture Microdissection
datasetposted on 2022-09-25, 21:03 authored by Jongmin Woo, Madeline Schoenfeld, Xinguo Sun, Thierry Iraguha, Zhanxiang Zhou, Qibin Zhang
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.
smooth muscle regionssimplified toluidine bluepot protein digestionlaser capture microdissectionfrozen intestinal tissuecomparable proteomic coverageassociated pathological changesintestinal tissue enrichedfirst proteomics studybased proteomics approachproteome level directlysecreting cells locatedmouse paneth cellenriched proteome enabledalso assessed usingcell equivalent tissuespaneth cellproteomics studieshighly enrichedpaneth cellsmay alsocrypt proteomestem cellsisolate cells3600 cells000 cellswell definedvitro smaller numbersmall intestinecrypts equivalentcrypt regionassisted oneantimicrobial peptidesantimicrobial peptide1300 proteins