Systemic splicing factor deficiency causes tissue-specific defects: a zebrafish model for retinitis pigmentosa†

Linder, Bastian; Dill, Holger; Hirmer, Anja; Brocher, Jan; Lee, Gek Ping; Mathavan, Sinnakaruppan; Bolz, Hanno J.; Winkler, Christoph; Laggerbauer, Bernhard; Fischer, Utz

doi:10.1093/hmg/ddq473

Cited by 59 publications

(69 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Preparation of embryo lysates for Western blotting was performed as described previously (Linder et al 2011). Adult tissues were homogenized in lysis buffer (50 mM Tris-HCl at pH 7.5, 150 mM NaCl, 1% Triton X-100) Quantitative analysis of the number of motor neuron cell bodies per body segment shows that the ctdsp2-MO-mediated rescue is statistically significant.…”

Section: Western Blottingmentioning

confidence: 99%

See 1 more Smart Citation

Intronic miR-26b controls neuronal differentiation by repressing its host transcript,ctdsp2

Dill

Linder²,

Fehr³

et al. 2012

Genes Dev.

Self Cite

121

107

View full text Add to dashboard Cite

Differentiation of neural stem cells (NSCs) to neurons requires the activation of genes controlled by the repressor element 1 (RE1) silencing transcription factor (REST)/neuron-restrictive silencer factor (NRSF) protein complex. Important components of REST/NRSF are phosphatases (termed RNA polymerase II C-terminal domain small phosphatases [CTDSPs]) that inhibit RNA polymerase II and suppress neuronal gene expression in NSCs. Activation of genes controlled by CTDSPs is required for neurogenesis, but how this is achieved is not fully understood. Here we show that ctdsp2 is a target of miR-26b, a microRNA that is encoded in an intron of the ctdsp2 primary transcript. This intrinsic negative feedback loop is inactive in NSCs because miR-26b biogenesis is inhibited at the precursor level. Generation of mature miR-26b is activated during neurogenesis, where it suppresses Ctdsp2 protein expression and is required for neuronal cell differentiation in vivo.

show abstract

Section: Western Blottingmentioning

confidence: 99%

“…Generation of digoxigenin-or fluorescein-labeled antisense RNA probes and whole-mount RNA ISH and was carried out as described recently (Linder et al 2011). Digoxigenin-labeled miRCURY LNA detection probes were obtained from Exiqon.…”

Section: Rna and Lna Ishmentioning

confidence: 99%

Intronic miR-26b controls neuronal differentiation by repressing its host transcript,ctdsp2

Dill

Linder²,

Fehr³

et al. 2012

Genes Dev.

Self Cite

121

107

View full text Add to dashboard Cite

show abstract

“…In the past years, particularly zebrafish has been used as a model for various human diseases ranging from cancer (Etchin et al, 2011) to neurodegenerative diseases (Paquet et al, 2010;Linder et al, 2011). Of interest, zebrafish has also turned out to be an excellent model for regeneration (Jopling et al, 2010;Kikuchi et al, 2010;Diep et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

A microinjection protocol for the generation of transgenic killifish (Species: Nothobranchius furzeri)

Hartmann

Englert

2012

Developmental Dynamics

View full text Add to dashboard Cite

Background: A challenge in age research is the absence of short-lived vertebrate model organisms. The turquoise killifish Nothobranchius furzeri has an exceptionally short lifespan of 4-10 months depending on the strain. Thus, it possesses the shortest known maximum lifespan of a vertebrate species that can be bred in captivity. Results: Here we show the successful introduction of DNA and RNA molecules into the one-cell embryo of N. furzeri. For this purpose, we adapted existing microinjection protocols to inject through the remarkably thick and robust chorion of N. furzeri's eggs. The injected DNA transgene was integrated into the genome and transmitted to subsequent generations as indicated by the expression of the fluorophore enhanced green fluorescent protein (EGFP). Furthermore, we could confirm a special phase during embryonic development in which embryogenesis occurs within a re-aggregated mass of previously dispersed cells as it has been described for other related cyprinodont fish species. Conclusions: The transgenesis protocol described here provides a basis for a variety of genetic manipulations including overexpression of genes and determining their effects on lifespan and longevity. The feasibility to perform transgenesis is an important step to establish N. furzeri as a new model in age research.

show abstract

“…42,43 In addition, tri-snRNP dysfunction, by silencing the systemic splicing factors prpf31 and prpf4, led to specific defects in retinal gene expression in a zebrafish model of retinitis pigmentosa, a hereditary eye disease that causes blindness due to a progressive loss of photoreceptors in the retina. 44 In vivo mutation of pre-mRNA processing factor 8 (Prpf8) in an ENU-induced zebrafish mutant, Cephalophonus, led to accumulation of aberrantly spliced transcripts retaining both U2-and U12-type introns that cause impaired myeloid differentiation in zebrafish. 45 In summary, our results suggest that Usb1 deficiency in zebrafish affects the splicing of specific transcripts that elicit defects in specific tissues, such as neutrophils.…”

Section: Discussionmentioning

confidence: 99%

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

Systemic splicing factor deficiency causes tissue-specific defects: a zebrafish model for retinitis pigmentosa†

Cited by 59 publications

References 33 publications

Intronic miR-26b controls neuronal differentiation by repressing its host transcript,ctdsp2

Intronic miR-26b controls neuronal differentiation by repressing its host transcript,ctdsp2

A microinjection protocol for the generation of transgenic killifish (Species: Nothobranchius furzeri)

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

Contact Info

Product

Resources

About