Open AccessImmunohistochemical analysis reveals variations in proteasome tissue expression in C. elegans
Author(s) -
Elisa Mikkonen,
Caj Haglund,
Carina I. Holmberg
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0183403
Subject(s) - proteasome , biology , gene knockdown , microbiology and biotechnology , ubiquitin , cell type , protein degradation , rna interference , multicellular organism , caenorhabditis elegans , intracellular , cell , cell culture , biochemistry , genetics , gene , rna
The ubiquitin-proteasome system (UPS) plays a crucial part in normal cell function by mediating intracellular protein clearance. We have previously shown that UPS-mediated protein degradation varies in a cell type-specific manner in C . elegans . Here, we use formalin-fixed, paraffin-embedded C . elegans sections to enable studies on endogenous proteasome tissue expression. We show that the proteasome immunoreactivity pattern differs between cell types and within subcellular compartments in adult wild-type (N2) C . elegans . Interestingly, widespread knockdown of proteasome subunits by RNAi results in tissue-specific changes in proteasome expression instead of a uniform response. In addition, long-lived daf-2(e1370) mutants with impaired insulin/IGF-1 signaling (IIS) display similar proteasome tissue expression as aged-matched wild-type animals. Our study emphasizes the importance of alternate approaches to the commonly used whole animal lysate-based methods to detect changes in proteasome expression occurring at the sub-cellular, cell or tissue resolution level in a multicellular organism.