Cell-Specific Inducible Gene Recombination in Postnatal Inner Ear Supporting Cells and Glia

Gómez-Casati, María Eugenia; Murtie, Joshua; Taylor, Brian F.; Corfas, Gabriel

doi:10.1007/s10162-009-0191-x

Cited by 51 publications

(50 citation statements)

References 28 publications

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“…1B). This same cell specificity was described previously using the Rosa26 LacZ reporter (27). Activation of the DTA transgene leads to cell-autonomous death via apoptosis (29)(30)(31).…”

Section: Significancementioning

confidence: 79%

“…To do so, we created mice expressing the tamoxifen-dependent Cre recombinase (CreER T ) under the control of the proteolipid protein 1 (Plp1) promoter (PlpCreER T ) (26,27) and carrying an inducible flox-stop DTA transgene (Rosa26 DTA ) (25) together with an inducible Tomato reporter (Ai14:Rosa26 tdTom ) (28). Tamoxifen treatment of neonatal [postnatal day (P)0-1] PlpCreER T ;Ai14:Rosa26 tdTom (PlpTom) mice resulted in specific and effective gene recombination in IBCs/IPhCs in all regions of the cochlea (apex, 25.9 ± 4.1%; middle, 71.9 ± 2.3%; base, 86.1 ± 4.2%; mean ± SEM; statistical differences exist only between the apex and the other cochlear turns) ( Fig.…”

Section: Significancementioning

confidence: 99%

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Spontaneous regeneration of cochlear supporting cells after neonatal ablation ensures hearing in the adult mouse

Lagarde

Wan

Zhang

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Supporting cells in the cochlea play critical roles in the development, maintenance, and function of sensory hair cells and auditory neurons. Although the loss of hair cells or auditory neurons results in sensorineural hearing loss, the consequence of supporting cell loss on auditory function is largely unknown. In this study, we specifically ablated inner border cells (IBCs) and inner phalangeal cells (IPhCs), the two types of supporting cells surrounding inner hair cells (IHCs) in mice in vivo. We demonstrate that the organ of Corti has the intrinsic capacity to replenish IBCs/IPhCs effectively during early postnatal development. Repopulation depends on the presence of hair cells and cells within the greater epithelial ridge and is independent of cell proliferation. This plastic response in the neonatal cochlea preserves neuronal survival, afferent innervation, and hearing sensitivity in adult mice. In contrast, the capacity for IBC/IPhC regeneration is lost in the mature organ of Corti, and consequently IHC survival and hearing sensitivity are impaired significantly, demonstrating that there is a critical period for the regeneration of cochlear supporting cells. Our findings indicate that the quiescent neonatal organ of Corti can replenish specific supporting cells completely after loss in vivo to guarantee mature hearing function.deafness | cell ablation | hair cell | organ of Corti | glia

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“…1B). This same cell specificity was described previously using the Rosa26 LacZ reporter (27). Activation of the DTA transgene leads to cell-autonomous death via apoptosis (29)(30)(31).…”

Section: Significancementioning

confidence: 79%

Section: Significancementioning

confidence: 99%

Spontaneous regeneration of cochlear supporting cells after neonatal ablation ensures hearing in the adult mouse

Lagarde

Wan

Zhang

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

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“…We also found a large number of Cre + Schwann cells in the spiral lamina (data not shown). There was no effect on morphology or cellular organization of the inner ear when BDNF was deleted using the Plp-CreER T2 allele and tamoxifen injections were given once a day from P5 to P11 (Gomez-Casati et al 2010).…”

Section: Cre/creer Lines For the Developing Vestibular Organsmentioning

confidence: 99%

Conditional Gene Expression in the Mouse Inner Ear Using Cre-loxP

Cox

Liu

Lagarde

et al. 2012

JARO

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In recent years, there has been significant progress in the use of Cre-loxP technology for conditional gene expression in the inner ear. Here, we introduce the basic concepts of this powerful technology, emphasizing the differences between Cre and CreER. We describe the creation and Cre expression pattern of each Cre and CreER mouse line that has been reported to have expression in auditory and vestibular organs. We compare the Cre expression patterns between Atoh1-CreER TM and Atoh1-CreER T2 and report a new line, Fgfr3-iCreER T2 , which displays inducible Cre activity in cochlear supporting cells. We also explain how results can vary when transgenic vs. knock-in Cre/CreER alleles are used to alter gene expression. We discuss practical issues that arise when using the Cre-loxP system, such as the use of proper controls, Cre efficiency, reporter expression efficiency, and Cre leakiness. Finally, we introduce other methods for conditional gene expression, including Flp recombinase and the tetracycline-inducible system, which can be combined with Cre-loxP mouse models to investigate conditional expression of more than one gene.

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“…For whole mount immunostaining, cristae were collected from adult Swiss Webster mice (Harlan Laboratories). For lineage tracing experiments, proteolipid protein (PLP)/ CreER;mTmG mice were generated by crossing heterozygous Plp-Cre-ER T2 mice (Doerflinger et al 2003;Gomez-Casati et al 2010; Jackson Laboratories strain 005975) with homozygous ROSA-mT/mG mice (Muzumdar et al 2007; Jackson Laboratories strain 007576). Mice were genotyped for Cre recombinase using DNA obtained from tail clips with the primers: forward 5′-aacattctcccaccgtcagt-3′ and reverse 5′-catttgggccagctaaaccat-3′ and for the mutant Rosa26 allele using the primers: wild-type forward 5′-ctctgctgcctcctggcttct-3′, wild-type reverse 5′-cgaggcggatcacaagcaata-3′, and mutant reverse 5′-tcaatgggcgggggtcgtt-3′.…”

Section: Animalsmentioning

confidence: 99%

“…This suggests that even though the adult explants do not survive as well in culture as younger explants, their survivability does not continue to decline with age, but stabilizes between at least P30 and P56-70. To label support cells, we used PLP/CreER mice expressing an inducible Cre recombinase under the PLP promoter crossed to mTmG reporter mice that express mTomato prior to recombination and mGFP after Cre-mediated recombination (Doerflinger et al 2003;Muzumdar et al 2007;Gomez-Casati et al 2010). Upon tamoxifen treatment, the support cells and Schwann cells that express the PLP transgene expressed GFP in a dose-dependent manner (Fig.…”

Section: New Hair Cells Arise Through Transdifferentiation Of Supportmentioning

confidence: 99%

Hair Cell Generation by Notch Inhibition in the Adult Mammalian Cristae

Slowik

Bermingham‐McDonogh

2013

JARO

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Balance disorders caused by hair cell loss in the sensory organs of the vestibular system pose a significant health problem worldwide, particularly in the elderly. Currently, this hair cell loss is permanent as there is no effective treatment. This is in stark contrast to nonmammalian vertebrates who robustly regenerate hair cells after damage. This disparity in regenerative potential highlights the need for further manipulation in order to stimulate more robust hair cell regeneration in mammals. In the utricle, Notch signaling is required for maintaining the striolar support cell phenotype into the second postnatal week. Notch signaling has further been implicated in hair cell regeneration after damage in the mature utricle. Here, we investigate the role of Notch signaling in the mature mammalian cristae in order to characterize the Notch-mediated regenerative potential of these sensory organs. For these studies, we used the γ-secretase inhibitor, N-[N-(3,5-difluorophenacetyl)-Lalanyl]-S-phenylglycine t-butyl ester (DAPT), in conjunction with a method we developed to culture cristae in vitro. In postnatal and adult cristae, we found that 5 days of DAPT treatment resulted in a downregulation of the Notch effectors Hes1 and Hes5 and also an increase in the total number of Gfi1 + hair cells. Hes5, as reported by Hes5-GFP, was downregulated specifically in peripheral support cells. Using lineage tracing with proteolipid protein (PLP)/CreER;mTmG mice, we found that these hair cells arose through transdifferentiation of support cells in cristae explanted from mice up to 10 weeks of age. These transdifferentiated cells arose without proliferation and were capable of taking on a hair cell morphology, migrating to the correct cell layer, and assembling what appears to be a stereocilia bundle with a long kinocilium. Overall, these data show that Notch signaling is active in the mature cristae and suggest that it may be important in maintaining the support cell fate in a subset of peripheral support cells.

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Cell-Specific Inducible Gene Recombination in Postnatal Inner Ear Supporting Cells and Glia

Cited by 51 publications

References 28 publications

Spontaneous regeneration of cochlear supporting cells after neonatal ablation ensures hearing in the adult mouse

Spontaneous regeneration of cochlear supporting cells after neonatal ablation ensures hearing in the adult mouse

Conditional Gene Expression in the Mouse Inner Ear Using Cre-loxP

Hair Cell Generation by Notch Inhibition in the Adult Mammalian Cristae

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