PUBLICATION
Zebrafish Galectin 3 binding protein (Lgals3bp) is the target antigen of the microglial 4C4 monoclonal antibody
- Authors
- Rovira, M., Miserocchi, M., Montanari, A., Hammou, L., Chomette, L., Pozo, J., Imbault, V., Bisteau, X., Wittamer, V.
- ID
- ZDB-PUB-221027-5
- Date
- 2022
- Source
- Developmental Dynamics : an official publication of the American Association of Anatomists 252(3): 400-414 (Journal)
- Registered Authors
- Wittamer, Valerie
- Keywords
- MAC-2BP, brain, lectin, macrophages, proteomics, zgc:112492
- MeSH Terms
-
- Animals
- Antibodies, Monoclonal*/chemistry
- Antibodies, Monoclonal*/metabolism
- Galectin 3/metabolism
- Macrophages/metabolism
- Mammals
- Microglia*/metabolism
- Zebrafish
- PubMed
- 36285351 Full text @ Dev. Dyn.
Citation
Rovira, M., Miserocchi, M., Montanari, A., Hammou, L., Chomette, L., Pozo, J., Imbault, V., Bisteau, X., Wittamer, V. (2022) Zebrafish Galectin 3 binding protein (Lgals3bp) is the target antigen of the microglial 4C4 monoclonal antibody. Developmental Dynamics : an official publication of the American Association of Anatomists. 252(3):400-414.
Abstract
Background Two decades ago, the fish-specific monoclonal antibody 4C4 was found to be highly reactive to zebrafish microglia, the macrophages of the central nervous system. This has resulted in 4C4 being widely used, in combination with available fluorescent transgenic reporters to identify and isolate microglia. However, the target protein of 4C4 remains unidentified, which represents a major caveat. In addition, whether the 4C4 expression pattern is strictly restricted to microglial cells in zebrafish has never been investigated.
Results Having demonstrated that 4C4 is able to capture its native antigen from adult brain lysates, we used immunoprecipitation/mass-spectrometry, coupled to recombinant expression analyses, to identify its target. The cognate antigen was found to be a paralog of Galectin 3 binding protein (Lgals3bpb), known as MAC2-binding protein in mammals. Notably, 4C4 did not recognize other paralogs, demonstrating specificity. Moreover, our data show that Lgals3bpb expression, while ubiquitous in microglia, also identifies leukocytes in the periphery, including populations of gut and liver macrophages.
Conclusions The 4C4 monoclonal antibody recognizes Lgals3bpb, a predicted highly glycosylated protein whose function in the microglial lineage is currently unknown. Identification of Lgals3bpb as a new pan-microglia marker will be fundamental in forthcoming studies using the zebrafish model. This article is protected by copyright. All rights reserved.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping