PUBLICATION

Brain transcriptome profile after CRISPR-induced ghrelin mutations in zebrafish

Authors
Blanco, A.M., Cortés, R., Bertucci, J.I., Soletto, L., Sánchez, E., Valenciano, A.I., Cerdá-Reverter, J.M., Delgado, M.J.
ID
ZDB-PUB-191102-36
Date
2019
Source
Fish physiology and biochemistry   46(1): 1-21 (Journal)
Registered Authors
Cerdá-Reverter, José Miguel
Keywords
CRISPR/Cas9, Fish, Ghrelinergic system, Knockdown, RNA-seq, Transcriptomic
MeSH Terms
  • Animals
  • Brain/physiology*
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Ghrelin/genetics*
  • Transcriptome
  • Zebrafish/genetics*
PubMed
31673996 Full text @ Fish Physiol. Biochem.
Abstract
Ghrelin (GRL) is a gut-brain hormone with a role in a wide variety of physiological functions in mammals and fish, which points out the ghrelinergic system as a key element for the appropriate biological functioning of the organism. However, many aspects of the multifunctional nature of GRL remain to be better explored, especially in fish. In this study, we used the CRISPR/Cas9 genome editing technique to generate F0 zebrafish in which the expression of grl is compromised. Then, we employed high-throughput mRNA sequencing (RNA-seq) to explore changes in the brain transcriptome landscape associated with the silencing of grl. The CRISPR/Cas9 technique successfully edited the genome of F0 zebrafish resulting in individuals with considerably lower levels of GRL mRNAs and protein and ghrelin O-acyl transferase (goat) mRNAs in the brain, intestine, and liver compared to wild-type (WT) zebrafish. Analysis of brain transcriptome revealed a total of 1360 differentially expressed genes (DEGs) between the grl knockdown (KD) and WT zebrafish, with 664 up- and 696 downregulated DEGs in the KD group. Functional enrichment analysis revealed that DEGs are highly enriched for terms related to morphogenesis, metabolism (especially of lipids), entrainment of circadian clocks, oxygen transport, apoptosis, and response to stimulus. The present study offers valuable information on the central genes and pathways implicated in functions of GRL, and points out the possible involvement of this peptide in some novel functions in fish, such as apoptosis and oxygen transport.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping