Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia.
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.
['Patil|Prakash|P|', 'Uechi|Tamayo|T|', 'Kenmochi|Naoya|N|']
D000818:Animals D002454:Cell Differentiation D004195:Disease Models, Animal D005095:Exoribonucleases D055785:Gene Knockdown Techniques D060172:Morpholinos D009503:Neutropenia D009504:Neutrophils D016376:Oligonucleotides, Antisense D012326:RNA Splicing D012868:Skin Abnormalities D015027:Zebrafish D029961:Zebrafish Proteins
splicing zebrafish poikiloderma with neutropenia PN 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 RNA disease U6 biogenesis USB1, U Six Biogenesis 1 hpf, hours post fertilization semi-quantitative reverse transcription and polymerase chain reaction small nuclear ribonucleoproteins snRNPs sqRT-PCR