PUBLICATION
Identification and expression analysis of the zebrafish orthologues of mammalian MAP1LC3 gene family
- Authors
- Ganesan, S., Moussavi Nik, S.H., Newman, M., Lardelli, M.
- ID
- ZDB-PUB-140723-3
- Date
- 2014
- Source
- Experimental cell research 328(1): 228-37 (Journal)
- Registered Authors
- Lardelli, Michael, Newman, Morgan
- Keywords
- Alzheimer's Disease, Autophagy, Chloroquine, Hypoxia, LC3, Rapamycin
- MeSH Terms
-
- Animals
- Autophagy/drug effects
- Blotting, Western
- Embryo, Nonmammalian/cytology
- Embryo, Nonmammalian/drug effects
- Embryo, Nonmammalian/metabolism*
- Embryonic Development*
- Gene Expression Regulation, Developmental*
- Immunosuppressive Agents/pharmacology
- In Situ Hybridization
- Microtubule-Associated Proteins/genetics
- Microtubule-Associated Proteins/metabolism*
- Phylogeny
- RNA, Messenger/genetics
- Real-Time Polymerase Chain Reaction
- Reverse Transcriptase Polymerase Chain Reaction
- Sirolimus/pharmacology
- Zebrafish/genetics
- Zebrafish/growth & development
- Zebrafish/metabolism*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 25051050 Full text @ Exp. Cell Res.
Citation
Ganesan, S., Moussavi Nik, S.H., Newman, M., Lardelli, M. (2014) Identification and expression analysis of the zebrafish orthologues of mammalian MAP1LC3 gene family. Experimental cell research. 328(1):228-37.
Abstract
Autophagy is the principle pathway in a cell involved in clearing damaged proteins and organelles. Therefore autophagy is necessary to maintain turnover balance of peptides and homeostasis. Autophagy occurs at basal levels under normal conditions but can be upregulated by chemical inducers or stress conditions. The zebrafish (Danio rerio) serves as a versatile tool to understand the function of genes implicated in autophagy. We report the identification of the zebrafish orthologue of mammalian genes MAP1LC3A (map1lc3a) and MAP1LC3B (map1lc3b) by phylogenetic and conserved synteny analysis and examine their expression during embryonic development. Both the zebrafish map1lc3a and map1lc3b genes show maternally contributed expression in early embryogenesis. However, levels of map1lc3a transcript steadily increase until at least 120 hours post fertilisation (hpf) while the levels of map1lc3b show a more variable pattern across developmental time. We have also validated the LC3II/LC3I immunoblot assay in the presence of chloroquine (a lysosomal proteolysis inhibitor). We found that the LC3II/LC3I ratio is significantly increased in the presence of sodium azide treatment supporting that hypoxia induces autophagy in zebrafish. This was supported by our qPCR assay that showed an increase in map1lc3a transcript levels in the presence of sodium azide. In contrast levels of map1lc3b transcripts were reduced in the presence of rapamycin but showed no significant difference in the presence of sodium azide. Our study thus identifies the zebrafish orthologues of MAP1LC3A & MAP1LC3B and supports the use of zebrafish for the interaction between hypoxia, development and autophagy.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping