ZFIN ID: ZDB-PUB-150325-2
slc7a6os Gene Plays a Critical Role in Defined Areas of the Developing CNS in Zebrafish
Benini, A., Cignarella, F., Calvarini, L., Mantovanelli, S., Giacopuzzi, E., Zizioli, D., Borsani, G.
Date: 2015
Source: PLoS One   10: e0119696 (Journal)
Registered Authors: Borsani, Giuseppe
Keywords: Embryos, Zebrafish, Gene expression, Central nervous system, RNA polymerase, Morpholino, Somites, Polymerase chain reaction
MeSH Terms:
  • Analysis of Variance
  • Animals
  • Base Sequence
  • Brain/embryology
  • Brain/metabolism*
  • Carrier Proteins/genetics
  • Central Nervous System/embryology*
  • Central Nervous System/enzymology*
  • Computational Biology
  • Gene Expression Regulation, Developmental/genetics
  • Gene Expression Regulation, Developmental/physiology*
  • Gene Expression Regulation, Enzymologic/genetics
  • Gene Expression Regulation, Enzymologic/physiology*
  • Gene Knockdown Techniques
  • In Situ Hybridization
  • Molecular Sequence Data
  • Nerve Tissue Proteins/genetics
  • Nerve Tissue Proteins/metabolism*
  • Peptide Hydrolases/genetics*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sequence Homology
  • Zebrafish*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed: 25803583 Full text @ PLoS One
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ABSTRACT
The aim of this study is to shed light on the functional role of slc7a6os, a gene highly conserved in vertebrates. The Danio rerio slc7a6os gene encodes a protein of 326 amino acids with 46% identity to human SLC7A6OS and 14% to Saccharomyces cerevisiae polypeptide Iwr1. Yeast Iwr1 specifically binds RNA pol II, interacts with the basal transcription machinery and regulates the transcription of specific genes. In this study we investigated for the first time the biological role of SLC7A6OS in vertebrates. Zebrafish slc7a6os is a maternal gene that is expressed throughout development, with a prevalent localization in the developing central nervous system (CNS). The gene is also expressed, although at different levels, in various tissues of the adult fish. To determine the functional role of slc7a6os during zebrafish development, we knocked-down the gene by injecting a splice-blocking morpholino. At 24 hpf morphants show morphological defects in the CNS, particularly the interface between hindbrain and midbrain is not well-defined. At 28 hpf the morpholino injected embryos present an altered somite morphology and appear partially or completely immotile. At this stage the midbrain, hindbrain and cerebellum are compromised and not well defined compared with control embryos. The observed alterations persist at later developmental stages. Consistently, the expression pattern of two markers specifically expressed in the developing CNS, pax2a and neurod, is significantly altered in morphants. The co-injection of embryos with synthetic slc7a6os mRNA, rescues the morphant phenotype and restores the wild type expression pattern of pax2a and neurod. Our data suggest that slc7a6os might play a critical role in defined areas of the developing CNS in vertebrates, probably by regulating the expression of key genes.
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