Transcript Profiling of Functionally Related Groups of Genes During Conditional Differentiation of a Mammalian Cochlear Hair Cell Line

Abstract

We have used Affymetrix high-density gene arrays to generate a temporal profile of gene expression during differentiation of UB/OC-1, a conditionally immortal cell line derived from the mouse cochlea. Gene expression was assessed daily for 14 days under differentiating conditions. The experiment was replicated in two separate populations of cells. Profiles for selected genes were correlated with those obtained by RT-PCR, TaqMan analysis, immunoblotting, and immunofluorescence. The results suggest that UB/OC-1 is derived from a population of nonsensory epithelial cells in the greater epithelial ridge that have the potential to differentiate into a hair-cell-like phenotype, without the intervention of Math1. Elements of the Notch signaling cascade were identified, including the receptor Notch3, with a transient up-regulation that suggests a role in hair cell differentiation. Several genes showed a profile similar to Notch3, including the transcriptional co-repressor Groucho1. UB/OC-1 also expressed Me1, a putative partner of Math1 that may confer competence to differentiate into hair cells. Cluster analysis revealed expression profiles for neural guidance genes associated with Gata3. The temporal dimension of this analysis provides a powerful tool to study genetic mechanisms that underlie the conversion of nonsensory epithelial cells into hair cells

Molecular characterization of conditionally immortalized cell lines derived from mouse early embryonic inner ear

“…Apparently, expression of Math-1 is either not essential for expression of certain HC genes, including myosin VIIa and Brn3.1, or is down-regulated early, at least in these cell lines. An identical situation was found in conditionally immortalized cells from the organ of Corti (UB/OC-1, Rivolta et al, 2002), which express numerous HC genes in the absence of Math-1. All HC phenotype markers were easily detected in the inner ear and in whole embryo tissues.…”

“…Under our MTA agreement with GI, Wyeth (who absorbed GI) was allowed to examine this manuscript for 30 days before its submission. Monoclonal anti-pan cytokeratin (Sigma C2562) was used at 1:100 (157 g/ ml; as in Rivolta et al, 2002).…”

Information for gene networks in inner ear development: A study centered on the transcription factor gata2

“…The temporal proWle for gata3 in OC-1 was previously analysed with the Mas v 5 software for AVymetrix GeneChips (Rivolta et al, 2002b). This proWle was re-calculated using gMOS and plotted against that for gata2 (Fig.…”

Hair cell regeneration: An exciting phenomenon . . . But will restoring hearing and balance be possible?

“…Microarray analysis has been used to study inner-ear genes in mice, rats, and chicks [94][95][96][97]. To gain insights into the genetic programs that govern the regenerative capacity of hair cells, researchers have compared gene expression profiles of sensory epithelia taken from chick utricles and basilar papillae (cochlea) [97].…”