Technical report: Laser microdissection and pressure catapulting is superior to conventional manual dissection for isolating pure spiral ganglion fractions from the cochlea

Torkos, Attila; Wissel, Kirsten; Warnecke, Athanasia; Lenarz, Thomas; Stöver, Timo

doi:10.1016/j.heares.2007.09.004

Cited by 2 publications

(4 citation statements)

References 18 publications

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“…Nefl encodes Nefl, which is expressed in developing and mature neurons, including SGNs (Torkos et al, 2008;Sun et al, 2018). A previous study has shown that the Nefl CreER/+ mouse FIGURE 5 | Hair cell protein expression and distortion product otoacoustic emission (DPOAE) study.…”

Section: Discussionmentioning

confidence: 99%

“…Nefl encodes Nefl, which is expressed in developing and mature neurons, including SGNs ( Torkos et al, 2008 ; Sun et al, 2018 ). A previous study has shown that the Nefl CreER /+ mouse strain shows neuronal-specific Cre activity in the central nervous system ( Rotolo et al, 2008 ).…”

Section: Discussionmentioning

confidence: 99%

“…Nefl encodes neurofilament light chain (Nefl), which is a major neuronal cytoskeleton component expressed in the soma, dendrites, and axon of developing and mature neurons, including the neurons along the auditory pathway ( Liem, 1993 ; Illing, 2001 ; Liu et al, 2004 ). In the cochlea, Nefl is expressed in SGNs, but not in other types of cells such as hair cells ( Torkos et al, 2008 ; Sun et al, 2018 ). When bred with mice expressing the Cre-inducible iDTR, the SGNs of Nefl Cre/ + ;iDTR +/– offspring were expected to be specifically damaged following DT treatment without interfering with hair cells.…”

Section: Introductionmentioning

confidence: 99%

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Generation of a Spiral Ganglion Neuron Degeneration Mouse Model

Komal

Singh

et al. 2021

Front. Cell Dev. Biol.

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Spiral ganglion neurons (SGNs) can be injured by a wide variety of insults. However, there still is a lack of degeneration models to specifically damage the SGNs without disturbing other types of cells in the inner ear. This study aims to generate an SGN-specific damage model using the Cre-LoxP transgenic mouse strains. The Cre-inducible diphtheria toxin receptor (iDTR+/+) knock-in mouse strain was crossed with a mouse strain with Cre activity specific to neurons (NeflCreER/CreER). Expression of the Cre-recombinase activity was evaluated using the reporter mouse strain Ai9 at pre-hearing, hearing onset, and post-hearing stages. Accordingly, heterozygous NeflCreER/+;iDTR+/– mice were treated with tamoxifen on postnatal days 1–5 (P1–5), followed by diphtheria toxin (DT) or vehicle injection on P7, P14, and P21 to evaluate the SGN loss. Robust tamoxifen-induced Cre-mediated Ai9 tdTomato fluorescence was observed in the SGN area of heterozygous NeflCreER/+;Ai9+/– mice treated with tamoxifen, whereas vehicle-treated heterozygote mice did not show tdTomato fluorescence. Compared to vehicle-treated NeflCreER/+;iDTR+/– mice, DT-treated NeflCreER/+;iDTR+/– mice showed significant auditory brainstem response (ABR) threshold shifts and SGN cell loss. Hair cell count and functional study did not show significant changes. These results demonstrate that the NeflCreER/CreER mouse strain exhibits inducible SGN-specific Cre activity in the inner ear, which may serve as a valuable SGN damage model for regeneration research of the inner ear.

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Section: Discussionmentioning

confidence: 99%

“…Nefl encodes Nefl, which is expressed in developing and mature neurons, including SGNs ( Torkos et al, 2008 ; Sun et al, 2018 ). A previous study has shown that the Nefl CreER /+ mouse strain shows neuronal-specific Cre activity in the central nervous system ( Rotolo et al, 2008 ).…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Generation of a Spiral Ganglion Neuron Degeneration Mouse Model

Komal

Singh

et al. 2021

Front. Cell Dev. Biol.

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“…RNA isolation and reverse transcription were performed as described [9]. Primers used for glyceraldehyde-3-phosphate dehydrogenase, gapdh , and type II iodothyronine deiodinase, dio2 , were identical to those previously tested and characterised [9,10]. We used HotStarTaq Plus DNA polymerase (Qiagen) and PCR conditions were: denaturation 94°C 3mins; amplification 94°C 1min/54°C 1min/72°C 1min (cycle varied); final extension 72°C 10mins.…”

Section: Methodsmentioning

confidence: 99%

Thyroxine treatments do not correct inner ear defects in tmprss1 mutant mice

Hanifa¹,

Scott²,

Crewther

et al. 2010

NeuroReport

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Complete deficiency of a member of the type II transmembrane serine protease family, tmprss1 (also known as hepsin), is associated with severe to profound hearing loss in mice and a gross enlargement of the tectorial membrane in the cochlea. Levels of thyroxine in these mice have been shown to be significantly lower compared to wild type controls. Since thyroxine is critical for inner ear development, we delivered thyroxine to these mice during the prenatal or postnatal stage of development. Both treatments could not ameliorate hearing loss or correct deformities in the tectorial membrane of these mutant mice, suggesting that a deficiency in tmprss1 affects thyroxine responsiveness in the inner ear in vivo.

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Technical report: Laser microdissection and pressure catapulting is superior to conventional manual dissection for isolating pure spiral ganglion fractions from the cochlea

Cited by 2 publications

References 18 publications

Generation of a Spiral Ganglion Neuron Degeneration Mouse Model

Generation of a Spiral Ganglion Neuron Degeneration Mouse Model

Thyroxine treatments do not correct inner ear defects in tmprss1 mutant mice

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