Auditory proteomics: Methods, accomplishments and challenges

Jamesdaniel, Samson; Salvi, Richard; Coling, Donald

doi:10.1016/j.brainres.2009.02.026

Cited by 12 publications

(7 citation statements)

References 51 publications

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“…Highly sophisticated LC–MS techniques combined with data-dependent acquisition is one of the most powerful proteomic methods. Even tiny amounts of complex protein samples can be applied, leading to hundreds (and even thousands) of identified and relatively quantified proteins in short times . This highly promising method can be used for research on inner ear fluids, which are available in limited sample sizes and heterogeneous composition of perilymph.…”

Section: Discussionsupporting

confidence: 68%

“…Even tiny amounts of complex protein samples can be applied, leading to hundreds (and even thousands) of identified and relatively quantified proteins in short times. 38 This highly promising method can be used for research on inner ear fluids, which are available in limited sample sizes and heterogeneous composition of perilymph. In this study, the protein composition of 41 human perilymph samples was analyzed in single analysis.…”

Section: ■ Discussionmentioning

confidence: 99%

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Proteome Analysis of Human Perilymph Using an Intraoperative Sampling Method

et al. 2017

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The knowledge about the etiology and pathophysiology of sensorineural hearing loss (SNHL) is still very limited. This study aims at the improvement of understanding different types of SNHL by proteome analysis of human perilymph. Sampling of perilymph was established during inner ear surgeries (cochlear implantation, vestibular schwannoma surgeries), and safety of the sampling method was determined by checking hearing threshold with pure-tone audiometry postoperatively. An in-depth shot-gun proteomics approach was performed to identify cochlear proteins and the individual proteome in perilymph of patients. This method enables the identification and quantification of protein composition of perilymph. The proteome of 41 collected perilymph samples with volumes of 1-12 μL was analyzed by data-dependent acquisition, resulting in overall 878 detected protein groups. At least 203 protein groups were solely identified in perilymph, not in reference samples (serum, cerebrospinal fluid), displaying a specific protein pattern for perilymph. Samples were grouped by patient's age and surgery type, leading to the identification of some proteins specific to particular subgroups. Proteins with different abundances between different sample groups were subjected to classification by gene ontology annotations. The identified proteins might serve as biomarkers to develop tools for noninvasive inner ear diagnostics and to elucidate molecular profiles of SNHL.

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Section: Discussionsupporting

confidence: 68%

Section: ■ Discussionmentioning

confidence: 99%

Proteome Analysis of Human Perilymph Using an Intraoperative Sampling Method

et al. 2017

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“…6 The present study has been designed to address the modulation of high molecular weight proteins in the same animal models of asthma and has been performed by using 2D-DIGE since this technique is recognized as being able to resolve complex biological samples. 7 In the present study, mice were characterized regarding main features of asthma and displayed a full asthmatic phenotype including bronchial hyperresponsiveness, eosinophilic inflammation, and bronchial remodeling. 2D-DIGE analysis performed on whole lung extracts of individual animals revealed, among other differentially expressed proteins, increased amounts of PDIA6, GRP78, Annexin A6, hnRPA3, and Enolase in lung parenchyma from mice exposed to allergens for 5 days.…”

Section: ' Discussionmentioning

confidence: 99%

“…We previously reported a proteomic study in a preclinical asthma model using SELDI-TOF-MS for the detection of low molecular weight (mostly <20 kDa) proteins . The present study has been designed to address the modulation of high molecular weight proteins in the same animal models of asthma and has been performed by using 2D-DIGE since this technique is recognized as being able to resolve complex biological samples …”

Section: Discussionmentioning

confidence: 99%

Potential Therapeutic Target Discovery by 2D-DIGE Proteomic Analysis in Mouse Models of Asthma

Calvo

Fillet

Renaut³

et al. 2011

J. Proteome Res.

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As asthma physiopathology is complex and not fully understood to date; it is expected that new key mediators are still to be unveiled in this disease. The main objective of this study was to discover potential new target proteins with a molecular weight >20 kDa by using two-dimensional differential in-gel electrophoresis (2D-DIGE) on lung parenchyma extracts from control or allergen-exposed mice (ovalbumin). Two different mouse models leading to the development of acute airway inflammation (5 days allergen exposure) and airway remodeling (10 weeks allergen exposure) were used. This experimental setting allowed the discrimination of 33 protein spots in the acute inflammation model and 31 spots in the remodeling model displaying a differential expression. Several proteins were then identified by MALDI-TOF/TOF MS. Among those differentially expressed proteins, PDIA6, GRP78, Annexin A6, hnRPA3, and Enolase display an increased expression in lung parenchyma from mice exposed to allergen for 5 days. Conversely, Apolipoprotein A1 was shown to be decreased after allergen exposure in the same model. Analysis on lung parenchyma of mice exposed to allergens for 10 weeks showed decreased calreticulin levels. Changes in the levels of those different mediators were confirmed by Western blot and immunohistochemical analysis. Interestingly, alveolar macrophages isolated from lungs in the acute inflammation model displayed enhanced levels of GRP78. Moreover, intratracheal instillation of anti-GRP78 siRNA in allergen-exposed animals led to a decrease in eosinophilic inflammation and bronchial hyperresponsiveness. This study unveils new mediators of potential importance that are up- and down-regulated in asthma. Among up-regulated mediators, GRP-78 appears as a potential new therapeutic target worthy of further investigations.

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“…However, in this report, for the first time to our knowledge, proteomic analysis has been successfully performed on FFCE microdissected human archival spiral ganglion tissue. Tandem spray MS provides the most powerful and sophisticated technology of all contemporary proteomics methods [19] and the results of this investigation demonstrate that the level of sensitivity in this approach to proteomic analysis is adequate for the evaluation of minute tissue samples in archival temporal bone sections. The number of proteins identified using this technology can be improved beyond the results reported in this study through optimizations in each step of the identification process, including the utilization of expanding bioinformatic databases.…”

Section: Ffce Tissue Lmd Mass Spectrometry Proteomics Analysismentioning

confidence: 94%