Auditory hair cell precursors immortalized from the mammalian inner ear

Rivolta, Marcelo N.; Grix, N; Lawlor, Patrick; Ashmore, Jonathan; Jagger, Daniel J.; Holley, Matthew C.

doi:10.1098/rspb.1998.0477

Cited by 99 publications

(86 citation statements)

References 50 publications

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“…49 Cells were maintained in high glucose DMEM containing 10% FBS and 50 U/ml g-interferon (Genzyme, Cambridge, MA). Cells were cultured under permissive conditions: 331C, 7% CO 2 in DMEM supplemented with 10% FBS.…”

Section: Cell Culture and Viabilitymentioning

confidence: 99%

Flunarizine induces Nrf2-mediated transcriptional activation of heme oxygenase-1 in protection of auditory cells from cisplatin

Kim

et al. 2006

Cell Death Differ

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We investigated the cytoprotective mechanisms of flunarizine in cisplatin-induced death of auditory cells. Concomitant with an increase in viability, treatment with flunarizine resulted in a marked dissociation of Nrf2/Keap1 and subsequent intranuclear translocation of Nrf2, which was mediated by PI3K-Akt signaling. Overexpression of Nrf2 protected cells from cisplatin along with transcriptional activation of ARE to generate heme oxygenase-1 (HO-1). Pretreatment with flunarizine predominantly increased the transcriptional activity of HO-1 among Nrf2-driven transcripts, including HO-1, NQO1, GCLC, GCLM, GSTl-1, and GSTA4. Furthermore, both pharmacological inhibition and siRNA transfection of HO-1 completely abolished the flunarizine-mediated protection of HEI-OC1 cells and the primary rat (P2) organ of Corti explants from cisplatin. These results suggest that Nrf2-driven transcriptional activation of ARE through PI3K-Akt signaling augments the generation of HO-1, which may be a critically important determinant in cellular response toward cisplatin and the cytoprotective effect of flunarizine against cisplatin.

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Section: Cell Culture and Viabilitymentioning

confidence: 99%

Flunarizine induces Nrf2-mediated transcriptional activation of heme oxygenase-1 in protection of auditory cells from cisplatin

Kim

et al. 2006

Cell Death Differ

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“…Both UB/UE-1 and the Kalinec OC cells also appear to demonstrate asymmetric cell divisions at the time of differentiation (Rivolta and Holley, 2002b;Kalinec et al, 1999), which may be one mechanism by which clonal cells develop divergent phenotypes. By contrast, the well-studied organ of Corti cell lines UB/OC-1 and UB/OC-2 demonstrate uniform phenotypes when studied by immunocytochemistry (Rivolta et al, 1998).…”

Section: Multiple Phenotypes In One Cellmentioning

confidence: 86%

“…Conditionally immortalized organ of Corti cells UB/ OC-2 express many hair cell markers under proliferative conditions, including myosin VIIa, ␣9 AChR, and Brn3.1, and continue to do so after differentiation (Rivolta et al, 1998), whereas another cell line, UB/OC-1, by contrast, up-regulates the same genes. Similarly, another HC maker, myosin VI, is strongly expressed by both cell lines in proliferative conditions and is unchanged after differentiation (Rivolta et al, 1998). Although these findings parallel our own results regarding expression of some HC genes, they may also reflect the later stage at which the cells were derived (E13).…”

Section: Expression Of Genes Found In Mature Hcsmentioning

confidence: 99%

Molecular characterization of conditionally immortalized cell lines derived from mouse early embryonic inner ear

Germiller¹,

Smiley²,

Ellis³

et al. 2004

Developmental Dynamics

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“…We use OC-K3 cells, but the steps detailed here are useful for any cell line or primary cell culture. Numerous cell lines are now available for those interested in cell biological processes of auditory system derived cells (21,22). …”

mentioning

confidence: 99%

Multiplexed Isobaric Tagging Protocols for Quantitative Mass Spectrometry Approaches to Auditory Research

Vetter¹,

Basappa²

2016

Methods in Molecular Biology

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Modern biologists have at their disposal a large array of techniques used to assess the existence and relative or absolute quantity of any molecule of interest in a sample. However, implementing most of these procedures can be a daunting task for the first time, even in a lab with experienced researchers. Just choosing a protocol to follow can take weeks while all of the nuances are examined and it is determined whether a protocol will (a) give the desired results, (b) result in interpretable and unbiased data, and (c) be amenable to the sample of interest. We detail here a robust procedure for labeling proteins in a complex lysate for the ultimate differential quantification of protein abundance following experimental manipulations. Following a successful outcome of the labeling procedure, the sample is submitted for mass spectrometric analysis, resulting in peptide quantification and protein identification. While we will concentrate on cells in culture, we will point out procedures that can be used for labeling lysates generated from other tissues, along with any minor modifications required for such samples. We will also outline, but not fully document, other strategies used in our lab to label proteins prior to mass spectrometric analysis, and describe under which conditions each procedure may be desirable. What is not covered in this chapter is anything but the most brief introduction to mass spectrometry (instrumentation, theory, etc.), nor do we attempt to cover much in the way of software used for post hoc analysis. These two topics are dependant upon one's resources, and where applicable, one's collaborators. We strongly encourage the reader to seek out expert advice on topics not covered here.

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Auditory hair cell precursors immortalized from the mammalian inner ear

Cited by 99 publications

References 50 publications

Flunarizine induces Nrf2-mediated transcriptional activation of heme oxygenase-1 in protection of auditory cells from cisplatin

Flunarizine induces Nrf2-mediated transcriptional activation of heme oxygenase-1 in protection of auditory cells from cisplatin

Molecular characterization of conditionally immortalized cell lines derived from mouse early embryonic inner ear

Multiplexed Isobaric Tagging Protocols for Quantitative Mass Spectrometry Approaches to Auditory Research

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