Histochemical and Fluorescent Analyses of Mitochondrial Integrity in Chick Auditory Neurons following Deafferentation

Karnes, Hope Elizabeth; Scaletty, Peter Nicholas; Durham, Dianne

doi:10.3766/jaaa.21.3.9

Cited by 6 publications

(4 citation statements)

References 93 publications

(154 reference statements)

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“…To allow quantified comparison between sides and survival groups, the optical density of each neuron was normalized to generate a z‐score via a method that has been used extensively to quantify changes in staining intensities at the individual cell level (Born and Rubel,1988; Garden et al,1994, 1995a, b; Hyson and Rubel,1995; Karnes et al,2010). Briefly, for each section, the mean and standard deviation of the optical density were calculated across all measured neurons in the left (intact) NM and used as standards to which the optical density of each individual neuron in the same section on that side of the brain or on the opposite side (deprived side) was normalized.…”

Section: Methodsmentioning

confidence: 99%

Osteogenic response to BMP‐2 of hMSCs grown on apatite‐coated scaffolds

Davis

Case

Miller

et al. 2011

Biotech & Bioengineering

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Osteoconductive materials play a critical role in promoting integration with surrounding bone tissue and resultant bone repair in vivo. However, the impact of 3D osteoconductive substrates coupled with soluble signals on progenitor cell differentiation is not clear. In this study, we investigated the influence of bone morphogenetic protein-2 (BMP-2) concentration on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) when seeded in carbonated apatite-coated polymer scaffolds. Mineralized scaffolds were more hydrophilic and adsorbed more BMP-2 compared to nonmineralized scaffolds. Changes in alkaline phosphatase (ALP) activity within stimulated hMSCs were dependent on the dose of BMP-2 and the scaffold composition. We detected more cell-secreted calcium on mineralized scaffolds at all time points, and higher BMP-2 concentrations resulted in increased ALP and calcium levels. RUNX2 and IBSP gene expression within hMSCs was affected by both substrate and soluble signals, SP7 by soluble factors, and SPARC by substrate-mediated cues. The present data indicate that a combination of apatite and BMP-2 do not simply enhance the osteogenic response of hMSCs, but act through multiple pathways that may be both substrate- and growth factor-mediated. Thus, multiple signaling strategies will likely be necessary to achieve optimal bone regeneration.

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Section: Methodsmentioning

confidence: 99%

Osteogenic response to BMP‐2 of hMSCs grown on apatite‐coated scaffolds

Davis

Case

Miller

et al. 2011

Biotech & Bioengineering

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“…in 1% ammonium sulfide (adapted from (Karnes et al, 2010)). Subsequent to the staining of each complex, sections were washed 3x 10 min in PBS before they were dehydrated for 4 min in 70% ETOH, 4 min in 96% ETOH, 10 min in 100% ETOH and 10 min in Xylol and immediately mounted with Eukitt.…”

Section: Immunohistochemistry (Ihc)mentioning

confidence: 99%

Mitochondrial impairments contribute to Spinocerebellar ataxia type 1 progression and can be ameliorated by the mitochondria-targeted antioxidant MitoQ

Stucki

Ruegsegger

Steiner

et al. 2016

Free Radical Biology and Medicine

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Mitochondrial impairments contribute to Spinocerebellar ataxia type 1 progression and can be ameliorated by the mitochondria-targeted antioxidant MitoQ, Free Radical Biology and Medicine, http://dx.doi.org/10.1016/j.freeradbiomed.2016.07.005 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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“…Active cell death is a physiological process important for normal development and tissue homeostasis as well as in cell removal in response to a variety of pathological conditions. Recent study provides evidence for cell death by an apoptotic mechanism in animal models of ototoxicity associated with abnormal expression of some proteases [34,35]. It is well known that caspase-3 is situated at a key junction in the process, mediating apoptotic cascade from the intrinsic and extrinsic activation pathways [36][37][38].…”

Section: Discussionmentioning

confidence: 99%

Salicylate Initiates Apoptosis in the Spiral Ganglion Neuron of Guinea Pig Cochlea by Activating Caspase-3

et al. 2011

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Salicylate-induced ototoxicity leading to sensorineural hearing loss (SNHL) and tinnitus is well documented. However, the exact mechanisms are poorly defined. Caspase-3 is a member of the class of effector caspases and has been activated in nearly every model of apoptosis. To examine its role in salicylate-induced injury, we subjected guinea pigs to treatment with a specific inhibitor zDEVD-FMK via the round window niche (RWN) followed by a systemic injection of salicylate at a dose of 200 mg · kg(-1) · d(-1) i.p. for 10 consecutive days. For those animals administered with salicylate, immunohistochemical studies revealed that caspase-3 was activated in the spiral ganglion neurons (SGNs) and method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) to identify neuronal apoptosis showed that fragmented nuclei were distributed in Rosenthal's canal. Topical administration of the zDEVD-FMK at a concentration of 500 mM blocked caspase-3 activation and had an effect in reducing the number of TUNEL-positive auditory neurons. In contrast, the inhibitor at a concentration of 125 or 250 mM caused no variation in the expression of activated caspase-3, or in the ratio of TUNEL-positive neurons. These results indicate that caspase-3 is a crucial mediator of apoptosis induced by salicylate in the primary auditory neuron in vivo, and suggest that the specific inhibitor at a relatively high concentration may be therapeutically beneficial in salicylate-induced apoptosis.

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Histochemical and Fluorescent Analyses of Mitochondrial Integrity in Chick Auditory Neurons following Deafferentation

Abstract: Taken together, our results demonstrate that a subset of deafferented NM neurons, presumably those that die, preferentially upregulates SDH, perhaps via the tricarboxylic acid (TCA) cycle. These same neurons undergo ATPase uncoupling and an eventual loss of Deltapsi(m).

Cited by 6 publications

References 93 publications

Osteogenic response to BMP‐2 of hMSCs grown on apatite‐coated scaffolds

Osteogenic response to BMP‐2 of hMSCs grown on apatite‐coated scaffolds

Mitochondrial impairments contribute to Spinocerebellar ataxia type 1 progression and can be ameliorated by the mitochondria-targeted antioxidant MitoQ

Salicylate Initiates Apoptosis in the Spiral Ganglion Neuron of Guinea Pig Cochlea by Activating Caspase-3

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