ZFIN ID: ZDB-PUB-150408-8
Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia
Patil, P., Uechi, T., Kenmochi, N.
Date: 2015
Source: RNA Biology   12: 426-434 (Journal)
Registered Authors: Kenmochi, Naoya, Uechi, Tamayo
Keywords: CHT, caudal haematopoietic tissue, CRISPR, clustered regularly interspaced short palindromic repeats, GFP, green fluorescent protein, MO, morpholino antisense oligo, MPN1, mutated in poikiloderma with neutropenia, PN, RNA disease, U6 biogenesis, USB1, U Six Biogenesis 1, hpf, hours post fertilization, poikiloderma with neutropenia, semi-quantitative reverse transcription and polymerase chain reaction, small nuclear ribonucleoproteins, snRNPs, splicing, sqRT-PCR, zebrafish
MeSH Terms:
  • Animals
  • Cell Differentiation/genetics*
  • Disease Models, Animal
  • Exoribonucleases/genetics*
  • Exoribonucleases/metabolism
  • Gene Knockdown Techniques
  • Morpholinos/metabolism
  • Neutropenia/genetics*
  • Neutropenia/metabolism
  • Neutropenia/pathology*
  • Neutrophils/metabolism*
  • Neutrophils/pathology
  • Oligonucleotides, Antisense/metabolism
  • RNA Splicing*
  • Skin Abnormalities/genetics*
  • Skin Abnormalities/metabolism
  • Skin Abnormalities/pathology*
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
PubMed: 25849198 Full text @ RNA Biol.
Poikiloderma with neutropenia (PN) is a rare inherited disorder characterized by poikiloderma, facial dysmorphism, pachyonychia, short stature and neutropenia. The molecular testing of PN patients has identified mutations in the C16orf57 gene, which encodes a protein referred to as USB1 (U Six Biogenesis 1). In this study, we developed a zebrafish model of PN by the microinjection of morpholino antisense oligos to suppress usb1 gene function. Severe morphological defects, including a bent tail, thin yolk extension and reduced body length, were predominant in the Usb1-suppressed embryos (morphants). We also observed significantly decreased number of neutrophils in the morphants by Sudan Black staining. Interestingly, the splicing of genes involved in neutrophil differentiation and development, such as mpx, ncf1, ela3l and npsn, was aberrant in the morphants. However, the splicing of haematopoietic precursors and erythroid-specific genes was unaltered. Importantly, the neutrophil defects were almost completely rescued by co-injection of ela3l mRNA, the most markedly affected gene in the morphants. Our study demonstrated a possible role of USB1 in modulating the tissue-specific gene splicing that eventually leads to the impaired development of neutrophils. This zebrafish model could serve as a valuable tool to investigate the causative role of USB1 in PN pathogenesis.