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Activation of PI3K signaling prevents aminoglycoside-induced hair cell death in the murine cochlea.

Jadali A, Kwan KY - Biol Open (2016)

Bottom Line: In aging animals, components for active PI3K signaling are present but decrease in hair cells.Hair cells with activated PI3K signaling were more resistant to aminoglycoside-induced hair cell death.These results indicate that increased PI3K signaling in hair cells promote survival and the PI3K signaling pathway is a target for preventing aminoglycoside-induced hearing loss.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA Stem Cell Research Center and Keck Center for Collaborative Neuroscience, Rutgers University, Piscataway, NJ 08854, USA.

No MeSH data available.


Related in: MedlinePlus

Ablation and visualization of PTEN knockout cochlear cells. (A) Diagram of the neural subset (NS) Cre recombinase transgene, a STOP-floxed tdTomato reporter and a PTEN conditional knockout allele in the PTEN cKO mouse line. (B-D) Immunofluorescence images for (B) MYO7A-, (C) tdTomato- and (D) phalloidin-labeled hair cells from the NS Cre tdTomato PTEN conditional allele (PTEN cKO) mouse. (E) Merged image of MYO7A-, tdTomato- and phalloidin-labeled cells for hair cell counts. (F) Diagram of cells in the sensory epithelia. Red cells represent tdTomato-expressing cells. Line represents an optical section acquired by confocal microscopy. (G) Percentage of tdTomato-expressing hair cells (HC) or surrounding cells (SC) (n=4) (H) Percentage of tdTomato-expressing IHC and OHC (n=4). Data represented as mean±s.d.
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BIO016758F5: Ablation and visualization of PTEN knockout cochlear cells. (A) Diagram of the neural subset (NS) Cre recombinase transgene, a STOP-floxed tdTomato reporter and a PTEN conditional knockout allele in the PTEN cKO mouse line. (B-D) Immunofluorescence images for (B) MYO7A-, (C) tdTomato- and (D) phalloidin-labeled hair cells from the NS Cre tdTomato PTEN conditional allele (PTEN cKO) mouse. (E) Merged image of MYO7A-, tdTomato- and phalloidin-labeled cells for hair cell counts. (F) Diagram of cells in the sensory epithelia. Red cells represent tdTomato-expressing cells. Line represents an optical section acquired by confocal microscopy. (G) Percentage of tdTomato-expressing hair cells (HC) or surrounding cells (SC) (n=4) (H) Percentage of tdTomato-expressing IHC and OHC (n=4). Data represented as mean±s.d.

Mentions: To validate the use of bpV(HOpic) for activating PI3K signaling, we employed a PTEN conditional knockout mouse model. Since bpV(HOpic) in the cochlear explants could affect both hair cell and supporting cell types, we used a neural-subset (NS) Cre recombinase mouse (Ljungberg et al., 2009) that targets both hair cells and supporting cells for Cre-mediated PTEN excision. The NS Cre line was generated using a fragment of the human GFAP promoter to drive Cre recombinase (Backman et al., 2001). To visualize and correlate Cre expression in the cochlea, the NS Cre mouse was mated to a red fluorescent protein (tdTomato) reporter mouse (Madisen et al., 2010). In these reporter mice, expression of Cre mediates excision of a loxP-flanked STOP cassette and allows for tdTomato expression. Expression of tdTomato correlates to Cre excision of the PTEN conditional knockout allele resulting in the loss of PTEN (Fig. 5A). We generated a NS Cre tdTomato PTEN conditional knockout mouse (PTEN cKO) to study the effects of upregulating PI3K signaling in the cochlea.Fig. 5.


Activation of PI3K signaling prevents aminoglycoside-induced hair cell death in the murine cochlea.

Jadali A, Kwan KY - Biol Open (2016)

Ablation and visualization of PTEN knockout cochlear cells. (A) Diagram of the neural subset (NS) Cre recombinase transgene, a STOP-floxed tdTomato reporter and a PTEN conditional knockout allele in the PTEN cKO mouse line. (B-D) Immunofluorescence images for (B) MYO7A-, (C) tdTomato- and (D) phalloidin-labeled hair cells from the NS Cre tdTomato PTEN conditional allele (PTEN cKO) mouse. (E) Merged image of MYO7A-, tdTomato- and phalloidin-labeled cells for hair cell counts. (F) Diagram of cells in the sensory epithelia. Red cells represent tdTomato-expressing cells. Line represents an optical section acquired by confocal microscopy. (G) Percentage of tdTomato-expressing hair cells (HC) or surrounding cells (SC) (n=4) (H) Percentage of tdTomato-expressing IHC and OHC (n=4). Data represented as mean±s.d.
© Copyright Policy - open-access
Related In: Results  -  Collection

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BIO016758F5: Ablation and visualization of PTEN knockout cochlear cells. (A) Diagram of the neural subset (NS) Cre recombinase transgene, a STOP-floxed tdTomato reporter and a PTEN conditional knockout allele in the PTEN cKO mouse line. (B-D) Immunofluorescence images for (B) MYO7A-, (C) tdTomato- and (D) phalloidin-labeled hair cells from the NS Cre tdTomato PTEN conditional allele (PTEN cKO) mouse. (E) Merged image of MYO7A-, tdTomato- and phalloidin-labeled cells for hair cell counts. (F) Diagram of cells in the sensory epithelia. Red cells represent tdTomato-expressing cells. Line represents an optical section acquired by confocal microscopy. (G) Percentage of tdTomato-expressing hair cells (HC) or surrounding cells (SC) (n=4) (H) Percentage of tdTomato-expressing IHC and OHC (n=4). Data represented as mean±s.d.
Mentions: To validate the use of bpV(HOpic) for activating PI3K signaling, we employed a PTEN conditional knockout mouse model. Since bpV(HOpic) in the cochlear explants could affect both hair cell and supporting cell types, we used a neural-subset (NS) Cre recombinase mouse (Ljungberg et al., 2009) that targets both hair cells and supporting cells for Cre-mediated PTEN excision. The NS Cre line was generated using a fragment of the human GFAP promoter to drive Cre recombinase (Backman et al., 2001). To visualize and correlate Cre expression in the cochlea, the NS Cre mouse was mated to a red fluorescent protein (tdTomato) reporter mouse (Madisen et al., 2010). In these reporter mice, expression of Cre mediates excision of a loxP-flanked STOP cassette and allows for tdTomato expression. Expression of tdTomato correlates to Cre excision of the PTEN conditional knockout allele resulting in the loss of PTEN (Fig. 5A). We generated a NS Cre tdTomato PTEN conditional knockout mouse (PTEN cKO) to study the effects of upregulating PI3K signaling in the cochlea.Fig. 5.

Bottom Line: In aging animals, components for active PI3K signaling are present but decrease in hair cells.Hair cells with activated PI3K signaling were more resistant to aminoglycoside-induced hair cell death.These results indicate that increased PI3K signaling in hair cells promote survival and the PI3K signaling pathway is a target for preventing aminoglycoside-induced hearing loss.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA Stem Cell Research Center and Keck Center for Collaborative Neuroscience, Rutgers University, Piscataway, NJ 08854, USA.

No MeSH data available.


Related in: MedlinePlus