Targeted mutation of the gene encoding prion protein in zebrafish reveals a conserved role in neuron excitability

Fleisch, Valerie C.; Leighton, Patricia L.A.; Wang, Hao; Pillay, Laura M.; Ritzel, R. Gary; Bhinder, Ganive; Roy, Birbickram; Tierney, Keith B.; Ali, Declan W.; Waskiewicz, Andrew J.; Allison, W. Ted

doi:10.1016/j.nbd.2013.03.007

Cited by 48 publications

(66 citation statements)

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“…We also observed evidence of NMD or loss of protein in selected mutants supporting the fact that many are likely null alleles, yet we still observed a high false-positive morphant phenotype rate. Consistent with these observations, recent studies have reported inconsistencies between morphants and mutants for individual genes, including phenotypes affecting lymphatic development (nr2f2, Aranguren et al, 2011; Swift et al, 2014; van Impel et al, 2014), neuronal function ( mfn2 and prp2 , Chapman et al, 2013; Fleisch et al, 2013; Nourizadeh-Lillabadi et al, 2010; Vettori et al, 2011), hindbrain patterning ( gbx2 , Kikuta et al, 2003; Su et al, 2014), and lateral line development ( tcf7 , Aman et al, 2011). Taken together with our broader findings here, these studies raise a major concern about relying solely on the application of MOs for primary characterization of gene function in the zebrafish.…”

Section: Discussionmentioning

confidence: 78%

Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish

et al. 2015

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SUMMARY The widespread availability of programmable site-specific nucleases now enables targeted gene disruption in the zebrafish. In this study, we applied site-specific nucleases to generate zebrafish lines bearing individual mutations in more than twenty genes. We found that mutations in only a small proportion of genes caused defects in embryogenesis. Moreover, mutants for ten different genes failed to recapitulate published Morpholino-induced phenotypes (morphants). The absence of phenotypes in mutant embryos was not likely due to maternal effects or failure to eliminate gene function. Consistently, a comparison of published morphant defects with the Sanger Zebrafish Mutation Project revealed that approximately eighty percent of morphant phenotypes were not observed in mutant embryos, similar to our mutant collection. Based on these results, we suggest that mutant phenotypes become the standard metric to define gene function in zebrafish, after which Morpholinos that recapitulate respective phenotypes could be reliably applied for ancillary analyses.

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Section: Discussionmentioning

confidence: 78%

Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish

et al. 2015

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“…Motion of larvae was recorded from above with larvae on the same computer monitor used for OMR (IBM 6734-HBO ThinkVision) similarly oriented (laid horizontally) and projecting a white screen. A video camera (Color CCTV Camera WV-CL930, Panasonic Tokyo) was mounted overhead connected to a PC running EthoVision ® XT7 software (Noldus, Wageningen, Netherlands), via a video capture card (Picolo H.264; Euresys, San Juan Capistrano, CA), as described previously [44]. Ethovision software was used to quantify the distance larvae travelled during the touch evoked escape response.…”

Section: Methodsmentioning

confidence: 99%

Rapid Recovery of Visual Function Associated with Blue Cone Ablation in Zebrafish

et al. 2016

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Hurdles in the treatment of retinal degeneration include managing the functional rewiring of surviving photoreceptors and integration of any newly added cells into the remaining second-order retinal neurons. Zebrafish are the premier genetic model for such questions, and we present two new transgenic lines allowing us to contrast vision loss and recovery following conditional ablation of specific cone types: UV or blue cones. The ablation of each cone type proved to be thorough (killing 80% of cells in each intended cone class), specific, and cell-autonomous. We assessed the loss and recovery of vision in larvae via the optomotor behavioural response (OMR). This visually mediated behaviour decreased to about 5% or 20% of control levels following ablation of UV or blue cones, respectively (P<0.05). We further assessed ocular photoreception by measuring the effects of UV light on body pigmentation, and observed that photoreceptor deficits and recovery occurred (p<0.01) with a timeline coincident to the OMR results. This corroborated and extended previous conclusions that UV cones are required photoreceptors for modulating body pigmentation, addressing assumptions that were unavoidable in previous experiments. Functional vision recovery following UV cone ablation was robust, as measured by both assays, returning to control levels within four days. In contrast, robust functional recovery following blue cone ablation was unexpectedly rapid, returning to normal levels within 24 hours after ablation. Ablation of cones led to increased proliferation in the retina, though the rapid recovery of vision following blue cone ablation was demonstrated to not be mediated by blue cone regeneration. Thus rapid visual recovery occurs following ablation of some, but not all, cone subtypes, suggesting an opportunity to contrast and dissect the sources and mechanisms of outer retinal recovery during cone photoreceptor death and regeneration.

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“…Conflicting data on the spatiotemporal expression pattern of PrP1 during embryonic development have been published (Cotto et al, 2005; Malaga-Trillo et al, 2009). The evidence is clearer for PrP2 expression, which is reportedly absent in the early embryo but reaches high levels in the nervous system during later developmental stages and is maintained following hatching (Malaga-Trillo et al, 2009; Fleisch et al, 2013). Knockdown of PrP1 or PrP2 expression using morpholinos has identified putative roles for both proteins in regulating cell adhesion (Malaga-Trillo et al, 2009; Huc-Brandt et al, 2014; Sempou et al, 2016).…”

Section: Animal Models For Investigating Prpc Functionmentioning

confidence: 99%

“…Knockdown of PrP1 or PrP2 expression using morpholinos has identified putative roles for both proteins in regulating cell adhesion (Malaga-Trillo et al, 2009; Huc-Brandt et al, 2014; Sempou et al, 2016). Furthermore, complete knockout of PrP2 expression using zinc finger nuclease technology increased susceptibility to seizures (Fleisch et al, 2013), suggesting that PrP2 might modulate neuronal excitability. There is some evidence that mammalian PrP C has similar functions to the zebrafish PrPs and this will be described in more detail in subsequent sections.…”

Section: Animal Models For Investigating Prpc Functionmentioning

confidence: 99%

“…Nevertheless, there are data from animal models other than mice that support the involvement of PrP C in the modulation of neuronal excitability. For example, knocking out PrP2 expression in zebrafish using zinc finger nuclease technology affected NMDAR currents and increased susceptibility to seizures induced by the drug pentylenetetrazol (Fleisch et al, 2013). Furthermore, in a study of the Drosophila neuromuscular junction, researchers found that murine PrP C expression resulted in enlarged synaptic vesicles and increased the probability of neurotransmitter release (Robinson et al, 2014).…”

Section: Prpc Functionmentioning

confidence: 99%

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Physiological Functions of the Cellular Prion Protein

Castle

Gill

2017

Front. Mol. Biosci.

169

157

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The prion protein, PrPC, is a small, cell-surface glycoprotein notable primarily for its critical role in pathogenesis of the neurodegenerative disorders known as prion diseases. A hallmark of prion diseases is the conversion of PrPC into an abnormally folded isoform, which provides a template for further pathogenic conversion of PrPC, allowing disease to spread from cell to cell and, in some circumstances, to transfer to a new host. In addition to the putative neurotoxicity caused by the misfolded form(s), loss of normal PrPC function could be an integral part of the neurodegenerative processes and, consequently, significant research efforts have been directed toward determining the physiological functions of PrPC. In this review, we first summarise important aspects of the biochemistry of PrPC before moving on to address the current understanding of the various proposed functions of the protein, including details of the underlying molecular mechanisms potentially involved in these functions. Over years of study, PrPC has been associated with a wide array of different cellular processes and many interacting partners have been suggested. However, recent studies have cast doubt on the previously well-established links between PrPC and processes such as stress-protection, copper homeostasis and neuronal excitability. Instead, the functions best-supported by the current literature include regulation of myelin maintenance and of processes linked to cellular differentiation, including proliferation, adhesion, and control of cell morphology. Intriguing connections have also been made between PrPC and the modulation of circadian rhythm, glucose homeostasis, immune function and cellular iron uptake, all of which warrant further investigation.

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Targeted mutation of the gene encoding prion protein in zebrafish reveals a conserved role in neuron excitability

Cited by 48 publications

References 66 publications

Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish

Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish

Rapid Recovery of Visual Function Associated with Blue Cone Ablation in Zebrafish

Physiological Functions of the Cellular Prion Protein

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