Analyses of pancreas development by generation of gfp transgenic zebrafish using an exocrine pancreas-specific elastaseA gene promoter

Wan, Honghui; Korzh, Svitlana; Li, Zhen; Mudumana, Sudha; Korzh, Vladimir; Jiang, Yun-Jin; Lin, Shuo; Gong, Zhiyuan

doi:10.1016/j.yexcr.2006.01.016

Cited by 83 publications

(73 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although we also observed partial rescue of morphological abnormalities, this might be due to the different roles of elastase in other tissues such as exocrine pancreas. [27][28][29] Overall, our results demonstrated that Usb1 deficiency in zebrafish recapitulates the PN phenotype with constant neutropenia that might have been derived from the similar mechanism of defective splicing of tissue-specific genes, as seen in Usb1-suppressed embryos. This presumption is due to the fact that haematopoietic differentiation and development in zebrafish closely resembles hematopoiesis in humans.…”

Section: Discussionsupporting

confidence: 64%

Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia

2015

View full text Add to dashboard Cite

Keywords: poikiloderma with neutropenia, RNA disease, splicing, U6 biogenesis, zebrafish Abbreviations: CHT, caudal haematopoietic tissue; CRISPR, clustered regularly interspaced short palindromic repeats; GFP, green fluorescent protein; hpf, hours post fertilization; MO, morpholino antisense oligo; MPN1, mutated in poikiloderma with neutropenia; PN; poikiloderma with neutropenia; snRNPs; small nuclear ribonucleoproteins; sqRT-PCR; semi-quantitative reverse transcription and polymerase chain reaction; USB1, U Six Biogenesis 1Poikiloderma 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.

show abstract

Section: Discussionsupporting

confidence: 64%

Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia

2015

View full text Add to dashboard Cite

show abstract

“…Expression of diphtheria toxin A-chain (DTA) in a tissuespecific manner can lead to the effective ablation of the target cell population, but it can also result in the undesired ablation of other cells, or even death of the organism, due to its potency 8,9 . This problem has prevented the establishment of stable zebrafish lines carrying the DTA transgene, so this method has only been used in transient transgenesis 9 . This technique could be used in an inducible manner if a suitable inducible promoter that is completely silent in the off-state could be identified.…”

Section: Existing Cell and Tissue Ablation Methodsmentioning

confidence: 99%

Nitroreductase-mediated cell/tissue ablation in zebrafish: a spatially and temporally controlled ablation method with applications in developmental and regeneration studies

2008

View full text Add to dashboard Cite

Ablation studies are used to elucidate cell lineage relationships, developmental roles for specific cells during embryogenesis and mechanisms of tissue regeneration. Previous chemical and genetic approaches to directed cell ablation have been hampered by poor specificity, limited efficacy, irreversibility, hypersensitivity to promoter leakiness, restriction to proliferating cells, slow inducibility or complex genetics. Here, we provide a step-by-step protocol for a hybrid chemicalgenetic cell ablation method in zebrafish that, by combining spatial and temporal control, is cell-type specific, inducible, reversible, rapid and scaleable. Bacterial Nitroreductase (NTR) is used to catalyze the reduction of the innocuous prodrug metrodinazole (Mtz), thereby producing a cytotoxic product that induces cell death. Based on this principle, NTR is expressed in transgenic zebrafish using a tissue-specific promoter. Subsequent exposure to Mtz by adding it to the media induces cell death exclusively within NTR + cells. This approach can be applied to regeneration studies, as removing Mtz by washing permits tissue recovery. Using this protocol, cell ablation can be achieved in 12-72 h, depending on the transgenic line used, and recovery initiates within the following 24 h.

show abstract

“…In contrast to hnf1ba hi2169 (I), Pdx1 expression (arrowheads) in the intestine and dorsal pancreatic islet (I) is not lost in hnf1ba zebrafish and mouse Hnf1b-null mutants exhibit a posterior expansion of hnf3b and sonic hedgehog (shh) expression into the presumptive duodenum (posterior foregut), the region from which the ventral and dorsal pancreas and liver develop (Sun and Hopkins, 2001;Haumaitre et al, 2005). Hedgehog signaling in the anterior endoderm is required early to pattern the anterior foregut via repression of more posterior endoderm fate (duodenum/pancreas/ liver) (Hebrok, 2003;Wan et al, 2006). Therefore, posterior expansion of shh expression would be expected to inhibit posterior foregut patterning events.…”

Section: Research Articlementioning

confidence: 99%

Specification of hepatopancreas progenitors in zebrafish by hnf1ba and wnt2bb

et al. 2013

View full text Add to dashboard Cite

SUMMARYAlthough the liver and ventral pancreas are thought to arise from a common multipotent progenitor pool, it is unclear whether these progenitors of the hepatopancreas system are specified by a common genetic mechanism. Efforts to determine the role of Hnf1b and Wnt signaling in this crucial process have been confounded by a combination of factors, including a narrow time frame for hepatopancreas specification, functional redundancy among Wnt ligands, and pleiotropic defects caused by either severe loss of Wnt signaling or Hnf1b function. Using a novel hypomorphic hnf1ba zebrafish mutant that exhibits pancreas hypoplasia, as observed in HNF1B monogenic diabetes, we show that hnf1ba plays essential roles in regulating β-cell number and pancreas specification, distinct from its function in regulating pancreas size and liver specification, respectively. By combining Hnf1ba partial loss of function with conditional loss of Wnt signaling, we uncover a crucial developmental window when these pathways synergize to specify the entire ventrally derived hepatopancreas progenitor population. Furthermore, our in vivo genetic studies demonstrate that hnf1ba generates a permissive domain for Wnt signaling activity in the foregut endoderm. Collectively, our findings provide a new model for HNF1B function, yield insight into pancreas and β-cell development, and suggest a new mechanism for hepatopancreatic specification.

show abstract

Analyses of pancreas development by generation of gfp transgenic zebrafish using an exocrine pancreas-specific elastaseA gene promoter

Cited by 83 publications

References 53 publications

Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia

Incomplete splicing of neutrophil-specific genes affects neutrophil development in a zebrafish model of poikiloderma with neutropenia

Nitroreductase-mediated cell/tissue ablation in zebrafish: a spatially and temporally controlled ablation method with applications in developmental and regeneration studies

Specification of hepatopancreas progenitors in zebrafish by hnf1ba and wnt2bb

Contact Info

Product

Resources

About