ABSTRACT: Inner ear auditory and vestibular tissues differ in their responses to mechanical stimuli. Chick cochlea and utricle sensory epithelia were microdissected at E20-E21. RNA was extracted and cRNA hybridized to Affymetrix microarrays.
Project description:Our work describes the application of mass spectrometry-based label-free quantitative proteomics for the identification of differentially expressed proteins between different-aged cochlea. We report proteins exclusively present as well as up- and downregulated in the cochlear sensory epithelium at significant developmental stages. Additionally, we report proteins exclusively present on P3 that were recently identified in the cochlea for the first time. We use bioinformatics to provide insight on biological functions and potential primary and secondary partners of the differentially expressed proteins. This study provides the first differentially expressed proteome in the mammalian cochlea at significant developmental stages; before hearing, during the onset of hearing, and when hearing is fully developed. We believe that these results will provide insights into the function of proteins that change during development and identify potential protein biomarkers related to hearing impairments.
Project description:Purified hair bundles and utricular epithelium from E19-E20 chick inner ears were analyzed by LC-MS/MS to determine the abundant and enriched proteins of the hair bundle. This dataset used a Thermo LTQ mass spectrometer for protein detection.
Project description:Purified hair bundles and utricular epithelium from E19-E20 chick inner ears were analyzed by LC-MS/MS to determine the abundant and enriched proteins of the hair bundle. This dataset used a Thermo LTQ Velos mass spectrometer for protein detection.
Project description:This is a reanalysis of PXD000104. Purified hair bundles and utricular epithelium from E19-E20 chick inner ears were analyzed by LC-MS/MS to determine the abundant and enriched proteins of the hair bundle. This dataset used a Thermo Orbitrap Elite mass spectrometer for protein detection.
Project description:This SuperSeries is composed of the following subset Series: GSE14783: Gallus gallus utricle striola vs. extra-striola GSE14784: GATA3 overexpression in utricle sensory epithelia GSE14785: GATA3 siRNA in utricle sensory epithelia Refer to individual Series
Project description:We report a gene expression changes during development and maturation of the murine utricle Overall design: Using RNA-sequencing, we examined the gene expression in the murine utricle at E17.5, p0, and p9
Project description:In order to gain insight into the molecular events which underlie auditory hair cell regeneration in chicken, we compared gene expression in chicken basilar papillae after 24, 48, and 72 hours in culture with or without forskolin (100uM).<br><br>Under sterile conditions, cochlear ducts containing the basilar papillae were carefully dissected out of ~4-day-old chicks and then individually cultured in DMEM with 10% fetal bovine serum with or without forskolin (final concentration 100 ?M, delivered in 1% DMSO) for either 24, 48, or 72 hours at 37ﾰC. Control samples received DMSO at 1% as a vehicle control. At the end of 3 days, the tegmentum vasculosum was dissected off to expose the auditory epithelium, which was delicately freed from the underlying cartilaginous plates. All explants were kept in culture for 3 days because new hair cells are first seen in basilar papillae treated with forskolin after ~3 days of exposure to the drug . Therefore, at even earlier time points in culture the molecular events that subserve hair cell proliferation are well underway. Each sample was comprised of 3 auditory epithelia from 3 different chicks which were put in ~100 ?L of DMEM and then immediately frozen at -80ﾰC until RNA isolation could be performed. There were a total of 24 samples in this experiment: six 72-hour forskolin, six 72-hour control, three 48-hour forskolin, three 48-hour control, three 24-hour forskolin, and three 24-hour control.
Project description:A comprehensive transcriptome profile of chicken utricle hair cell across a 7 days regenerative time course. Compare gene expression changes in chicken utricle treated with streptomycin to the control. A total of 60 samples were collected from 11 different time points. Each time point contains at least 2 biological samples.
Project description:Single-cell proteomics can reveal the changing protein composition of differentiating cells. We used shotgun mass spectrometry to determine the abundant proteins present in single or small pools of subpicoliter-sized cells from the embryonic day 15 (E15) utricle of the chicken inner ear, when many hair cells are differentiating from progenitor (supporting) cells. The actin monomer binding protein thymosin β4 (TMSB4X) was present in E15 progenitor cells at nearly equimolar levels relative to actin, but dropped to one-tenth that value in hair cells, with little change in total actin. Single-cell RNA-seq analysis of E15 utricle cells showed that TMSB4X transcripts fell in abundance once hair-cell differentiation initiated. These results suggest that most actin is sequestered in progenitor cells, but upon differentiation to hair cells, actin is released, permitting assembly of the sensory hair bundle.
Project description:The inner ear utilizes sensory hair cells as mechano-electric transducers for sensing sound and balance. In mammals, these hair cells lack the capacity for regeneration. Unlike mammals, hair cells from non-mammalian vertebrates, such as birds, can be regenerated throughout the life of the organism making them a useful model for studying inner ear genetics pathways. The zinc finger transcription factor GATA3 is required for inner ear development and mutations cause sensory neural deafness in humans. In the avian cochlea GATA3 is expressed throughout the sensory epithelia; however, expression is limited to the striola of the utricle. The striola corresponds to an abrupt change in morphologically distinct hair cell types and a 180° shift in hair cell orientation. We used 3 complimentary approaches to identify potential downstream targets of GATA3 in the avian utricle. Specifically we used microarray expression profiling of GATA3 knockdown by siRNA and GATA3 over-expression treatments as well as direct comparisons of GATA3 expressing cells from the striola and non GATA3 expressing cells from the extra-striola. Whole utricle specimens were treated with streptomycin for 24 hrs, rinsed and allowed to recover for an additional 24 hrs. Whole utricles were transfected with either GATA3 or GFP 21mer synthetic siRNAs for an additional 48 hrs and pure sensory epithelia were isolated. There are 2 biological samples and experiments include technical replicates as well as dye-switches for a total of 8 microarrays.