PUBLICATION

CTP synthase knockdown during early development distorts the nascent vertebral column and causes fluid retention in multiple tissues in zebrafish

Authors
Dzaki, N., Wahab, W., Azlan, A., Azzam, G.
ID
ZDB-PUB-180923-1
Date
2018
Source
Biochemical and Biophysical Research Communications   505(1): 106-112 (Journal)
Registered Authors
Keywords
CTP synthase, DON treatment, Development, Zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Body Fluids/metabolism*
  • Carbon-Nitrogen Ligases/classification
  • Carbon-Nitrogen Ligases/genetics*
  • Carbon-Nitrogen Ligases/metabolism
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/metabolism
  • Gene Expression Regulation, Developmental*
  • Gene Knockdown Techniques
  • Isoenzymes/classification
  • Isoenzymes/genetics
  • Isoenzymes/metabolism
  • Phylogeny
  • Sequence Homology, Amino Acid
  • Spine/embryology
  • Spine/metabolism*
  • Time Factors
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
PubMed
30241946 Full text @ Biochem. Biophys. Res. Commun.
Abstract
CTP Synthase (CTPS) is a metabolic enzyme that is recognized as a catalyst for nucleotide, phospholipid and sialoglycoprotein production. Though the structural characteristics and regulatory mechanisms of CTPS are well-understood, little is known regarding the extent of its involvement during the early developmental stages of vertebrates. Zebrafish carries two CTPS genes, annotated as ctps1a and ctps1b. Phylogenetic analyses show that both genes had diverged from homologues in the ancestral Actinopterygii, Oreochromis niloticus. Conservation of common CTPS-catalytic regions further establishes that both proteins are likely to be functionally similar to hsaCTPS. Here, we show that ctps1a is more critical throughout the initial period of embryonic development than ctps1b. The effects of concurrent partial knockdown are dependent on ctps1a vs ctps1b dosage ratios. When these are equally attenuated, abnormal phenotypes acquired prior to the pharyngula period disappear in hatchlings (48hpf); however, if either gene is more attenuated than the other, these only become more pronounced in advanced stages. Generally, disruption to normal ctps1a or ctps1b expression levels by morpholinos culminates in the distortion of the early spinal column as well as multiple-tissue oedema. Other effects include slower growth rates, increased mortality rates and impaired structural formation of the young fish's extremities. Embryos grown in DON, a glutamine-analogue drug and CTPS antagonist, also exhibit similar characteristics, thus strengthening the validity of the morpholino-induced phenotypes observed. Together, our results demonstrate the importance of CTPS for the development of zebrafish embryos, as well as a disparity in activity and overall importance amongst isozymes.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping