An improved fixation method for guinea pig cochlear tissues

Groot, J. C. M. J. De; Veldman, Jan E.; Huizing, Egbert H.

doi:10.3109/00016488709107323

Cited by 48 publications

(30 citation statements)

References 14 publications

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“…Bullfrog sympathetic ganglia were fixed in situ with a trialdehyde fixative (3% paraformaldehyde, 2% glutaraldehyde, and 1% acrolein; DeGroot et al, 1987) made in frog Ringer's solution. Besides the aldehydes, the final fixative solution contained the following (in mM): NaCl 115, KCl 2, CaCl 2 1.8, and HEPES 2 at pH 7.25.…”

Section: Materials and Methods Tissuementioning

confidence: 99%

Dense-cored vesicles, smooth endoplasmic reticulum, and mitochondria are closely associated with non-specialized parts of plasma membrane of nerve terminals: Implications for exocytosis and calcium buffering by intraterminal organelles

et al. 1999

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Section: Materials and Methods Tissuementioning

confidence: 99%

Dense-cored vesicles, smooth endoplasmic reticulum, and mitochondria are closely associated with non-specialized parts of plasma membrane of nerve terminals: Implications for exocytosis and calcium buffering by intraterminal organelles

et al. 1999

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“…Cochleae were decalcified in the same decalcifying solution for 5 days. All specimens were postfixed in a 1% aqueous OsO 4 solution containing 1% K 4 Ru(CN) 6 for 2 h at 4 ° C followed by several rinses in distilled water [De Groot et al, 1987]. Dehydration was performed in a graded ethanol and propylene oxide series.…”

Section: Tissue Processing For Histologymentioning

confidence: 99%

“…A hole was made in the apex of the cochlea, both the oval and round window membranes were perforated and the cochlea and vestibular labyrinth were fixed by intralabyrinthine perfusion with a tri-aldehyde fixative consisting of 3% glutaraldehyde, 2% formaldehyde, 1% acrolein, and 2.5% dimethylsulfoxide in 0.08 M sodium cacodylate buffer (pH 7.4) [De Groot et al, 1987]. Perfusion was followed by immersion in the same fixative for 3 h at room temperature, after which the bulla was rinsed several times in 0.1 M sodium cacodylate buffer (pH 7.4) and stored in the same buffer at 4 ° C. In order to facilitate the localization of the utricle and saccule during microdissection, an aqueous solution of 1% OsO 4 containing 1% K 4 Ru(CN) 6 [De Groot et al, 1987] was gently perfused through the hole in the apex of the cochlea, followed by immersion in the same solution for 20 min at 4 ° C.…”

Section: Tissue Processing For Histologymentioning

confidence: 99%

Combined Administration of Kanamycin and Furosemide Does Not Result in Loss of Vestibular Function in Guinea Pigs

Bremer

Groot²,

Versnel³

et al. 2011

Audiol Neurotol

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Aminoglycoside antibiotics are known to damage the vestibular and auditory sensory epithelia. Although loop diuretics enhance the cochleotoxic effect of aminoglycosides, it is not known whether concomitant administration of an aminoglycoside and a loop diuretic affects the vestibular system. The aim of our study was to investigate the effect of co-administration of kanamycin and furosemide upon the otolith organs and to compare it to the known vestibulotoxic effect of gentamicin. Five guinea pigs were injected with a single dose of both kanamycin (400 mg/kg, s.c.) and furosemide (100 mg/kg, i.v.), 5 animals received gentamicin (100 mg/kg, i.p.) for 10 days, and 5 untreated animals served as controls. After 7 days, vestibular function was assessed by measuring vestibular short-latency evoked potentials (VsEPs) to linear acceleration stimuli and cochlear function by auditory brainstem responses (ABRs) to clicks. Hair cell densities were determined in phalloidin-stained whole mounts of the utricles and saccules, and in midmodiolar sections of resin-embedded cochleae. Co-administration of kanamycin and furosemide had no significant effect on VsEPs and hair cell densities in the utricles and saccules were not reduced. ABR thresholds were increased to a great extent (by ∼60 dB), and histologically a severe loss of cochlear hair cells was observed. The effect of gentamicin, both on vestibular and cochlear function, was just the opposite. VsEP thresholds to horizontal stimulation were elevated and suprathreshold amplitudes showed a decrease, whereas cochlear function was not reduced. With this protocol, we have a tool to selectively induce cochlear or vestibular damage, which may be of interest to researchers and clinicians alike.

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“…Cochleas were processed as described in detail by De Groot et al (1987). Cochleas were embedded in toto in Spurr's low-viscosity resin and divided into two halves along a standardized midmodiolar plane (Van Ruijven et al, 2004).…”

Section: Histologymentioning

confidence: 99%

Time course of cochlear electrophysiology and morphology after combined administration of kanamycin and furosemide

et al. 2007

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In animal models of deafness, administration of an aminoglycoside in combination with a loop diuretic is often applied to produce a rapid loss of cochlear hair cells. However, the extent to which surviving hair cells remain functional after such a deafening procedure varies. In a longitudinal electrocochleographical study, we investigated the variability of cochlear function between and within guinea pigs after combined administration of kanamycin and furosemide. Concurrently, histological data were obtained at 1, 2, 4 and 8 weeks after deafening treatment. The main measures in our study were compound action potential (CAP) thresholds, percentage of surviving hair cells and packing density of spiral ganglion cells (SGCs). One day after deafening treatment, we found threshold shifts widely varying among animals from 0 to 100 dB. The variability decreased after 2 days, and in 18 out of 20 animals threshold shifts greater than 55 dB were found 4-7 days after deafening. Remarkably, in the majority of animals, thresholds decreased by up to 25 dB after 7 days indicating functional recovery. As expected, final thresholds were negatively correlated to the percentage of surviving hair cells. Notably, the percentage of surviving hair cells might be predicted on the basis of thresholds observed one day after deafening. SGC packing density, which rapidly decreased with the period after deafening treatment and correlated to the percentage of surviving inner hair cells, was not a determining factor for the CAP thresholds.

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An improved fixation method for guinea pig cochlear tissues

Cited by 48 publications

References 14 publications

Dense-cored vesicles, smooth endoplasmic reticulum, and mitochondria are closely associated with non-specialized parts of plasma membrane of nerve terminals: Implications for exocytosis and calcium buffering by intraterminal organelles

Dense-cored vesicles, smooth endoplasmic reticulum, and mitochondria are closely associated with non-specialized parts of plasma membrane of nerve terminals: Implications for exocytosis and calcium buffering by intraterminal organelles

Combined Administration of Kanamycin and Furosemide Does Not Result in Loss of Vestibular Function in Guinea Pigs

Time course of cochlear electrophysiology and morphology after combined administration of kanamycin and furosemide

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