Transgenic mice expressing the Peripherin-EGFP genomic reporter display intrinsic peripheral nervous system fluorescence

McLenachan, Samuel; Goldshmit, Yona; Fowler, Kerry J.; Voullaire, Lucille; Holloway, Timothy P.; Turnley, Ann M.; Ioannou, Panos; Sarsero, Joseph P.

doi:10.1007/s11248-008-9210-7

Cited by 15 publications

(19 citation statements)

References 43 publications

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“…We hypothesized that defective pruning of type II SGN fibers could account for the abnormal retention of presynaptic ribbons in Pit1 dw mutants. We took advantage of the Peripherin-GFP (PGFP) transgenic mouse model that has the protein peripherin, a commonly used marker for type II SGNs, labeled with GFP (McLenachan et al, 2008). The breeding of PGFP mice with Pit1 dw mice allowed a specific detection of type II SGNs in both WT and Pit1 dw cochleas from P6 onward (Figure 2A,B).…”

Section: Resultsmentioning

confidence: 99%

Thyroid hormone is required for the pruning of afferent type II spiral ganglion neurons in the mouse cochlea

2016

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Afferent connections to the sensory inner and outer hair cells in the cochlea refine and functionally mature during the thyroid hormone (TH)- critical period of inner ear development that occurs perinatally in rodents. In this study, we investigated the effects of hypothyroidism on afferent type II innervation to outer hair cells (OHCs) using the Snell dwarf mouse (Pit1dw). Using a transgenic approach to specifically label type II spiral ganglion neurons, we found that a lack of TH causes persistence of excess type II SGN connections to the OHCs, as well as continued expression of the hair cell functional marker, otoferlin, in the OHCs beyond the maturation period. We also observed a concurrent delay in efferent attachment to the OHCs. Supplementing with TH during the early postnatal period from postnatal day (P) 3 to P4 reversed the defect in type II SGN pruning but did not alter otoferlin expression. Our results show that hypothyroidism causes a defect in the large-scale pruning of afferent type II spiral ganglion neurons in the cochlea, and a delay in efferent attachment and the maturation of otoferlin expression. Our data suggest that the state of maturation of hair cells, as determined by otoferlin expression, may not regulate the pruning of their afferent innervation.

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Section: Resultsmentioning

confidence: 99%

Thyroid hormone is required for the pruning of afferent type II spiral ganglion neurons in the mouse cochlea

2016

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“…Prph is an interesting candidate because in mammals and zebrafish, it is only expressed in neurons (Fuchs and Weber, 1994; McLenachan et al, 2008), while Xenopus Prph is expressed in neurons, proliferating cells as well as glia (Gervasi et al, 2000). In mammals, peripherin is abundant in PNS neurons and is also expressed in RGCs (McLenachan et al, 2008; Wang et al, 2007). The peripherin response to retinal injury has not been determined in mammals.…”

Section: Discussionmentioning

confidence: 99%

Müller glia reactivity follows retinal injury despite the absence of the glial fibrillary acidic protein gene in Xenopus

Luna

Aruck

et al. 2017

Developmental Biology

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Intermediate filament proteins are structural components of the cellular cytoskeleton with cell-type specific expression and function. Glial fibrillary acidic protein (GFAP) is a type III intermediate filament protein and is up-regulated in glia of the nervous system in response to injury and during neurodegenerative diseases. In the retina, GFAP levels are dramatically increased in Müller glia and are thought to play a role in the extensive structural changes resulting in Müller cell hypertrophy and glial scar formation. In spite of similar changes to the morphology of Xenopus Müller cells following injury, we found that Xenopus lack a gfap gene. Other type III intermediate filament proteins were induced following rod photoreceptor ablation and retinal ganglion cell axotomy. The recently available X. tropicalis and X. laevis genomes indicate a small deletion most likely resulted in the loss of the gfap gene during evolution. Lastly, a survey of representative species from all three extant amphibian orders including the Anura (frogs, toads), Caudata (salamanders, newts), and Gymnophiona (caecilians) suggests that deletion of the gfap locus occurred after the common ancestor of the Anura and Caudata. Our results demonstrate that extensive changes in Müller cell morphology following retinal injury do not require GFAP in Xenopus, and other type III intermediate filament proteins may be involved in the gliotic response.

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“…As EGFP is one of the most commonly used reporter proteins in mouse transgenics owing to its enhanced photostability and strong fluorescence intensity [20], [21], [22], it is necessary to study its compatibility with the IRES- and F2A-based vector systems. Through the analysis of six locus-specific EGFP knock-in mouse lines that we have generated – Sox9 IRES-EGFP , Sox9 F2A-EGFP , FLAG 3 -Bapx1 F2A-EGFP , Bapx1 IRES-Cre-IRES-EGFP , Bapx1 F2A-Cre-F2A-EGFP and Bapx1 EGFP (hereafter these mouse lines will be denoted by their abbreviated forms as summarized in Table 1), we have demonstrated that both the IRES and F2A reliably co-expressed the concatenated proteins.…”

Section: Introductionmentioning

confidence: 99%

Comparison of IRES and F2A-Based Locus-Specific Multicistronic Expression in Stable Mouse Lines

et al. 2011

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Efficient and stoichiometric expression of genes concatenated by bi- or multi-cistronic vectors has become an invaluable tool not only in basic biology to track and visualize proteins in vivo, but also for vaccine development and in the clinics for gene therapy. To adequately compare, in vivo, the effectiveness of two of the currently popular co-expression strategies - the internal ribosome entry site (IRES) derived from the picornavirus and the 2A peptide from the foot-and-mouth disease virus (FDMV) (F2A), we analyzed two locus-specific knock-in mouse lines co-expressing SRY-box containing gene 9 (Sox9) and enhanced green fluorescent protein (EGFP) linked by the IRES (Sox9IRES-EGFP) or the F2A (Sox9F2A-EGFP) sequence. Both the constructs expressed Sox9 and EGFP proteins in the appropriate Sox9 expression domains, with the IRES construct expressing reduced levels of EGFP compared to that of the F2A. The latter, on the other hand, produced about 42.2% Sox9-EGFP fusion protein, reflecting an inefficient ribosome ‘skipping’ mechanism. To investigate if the discrepancy in the ‘skipping’ process was locus-dependent, we further analyzed the FLAG3-Bapx1F2A-EGFP mouse line and found similar levels of fusion protein being produced. To assess if EGFP was hindering the ‘skipping’ mechanism, we examined another mouse line co-expressing Bagpipe homeobox gene 1 homolog (Bapx1), Cre recombinase and EGFP (Bapx1F2A-Cre-F2A-EGFP). While the ‘skipping’ was highly efficient between Bapx1 and Cre, the ‘skipping’ between Cre and EGFP was highly inefficient. We have thus demonstrated in our comparison study that the efficient and close to equivalent expression of genes linked by F2A is achievable in stable mouse lines, but the EGFP reporter may cause undesirable inhibition of the ‘skipping’ at the F2A sequence. Hence, the use of other reporter genes should be explored when utilizing F2A peptides.

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Transgenic mice expressing the Peripherin-EGFP genomic reporter display intrinsic peripheral nervous system fluorescence

Cited by 15 publications

References 43 publications

Thyroid hormone is required for the pruning of afferent type II spiral ganglion neurons in the mouse cochlea

Thyroid hormone is required for the pruning of afferent type II spiral ganglion neurons in the mouse cochlea

Müller glia reactivity follows retinal injury despite the absence of the glial fibrillary acidic protein gene in Xenopus

Comparison of IRES and F2A-Based Locus-Specific Multicistronic Expression in Stable Mouse Lines

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