Differential display and gene arrays to examine auditory plasticity.
Differential gene expression forms the basis for development, differentiation, regeneration, and plasticity of tissues and organs. We describe two methods to identify differentially expressed genes. Differential display, a PCR-based approach, compares the expression of subsets of genes under two or more conditions. Gene arrays, or DNA microarrays, contain cDNAs from both known genes and novel genes spotted on a solid support (nylon membranes or glass slides). Hybridization of the arrays with RNA isolated from two different experimental conditions allows the simultaneous analysis of large numbers of genes, from hundreds to thousands to whole genomes. Using differential display to examine differential gene expression after noise trauma in the chick basilar papilla, we identified the UBE3B gene that encodes a new member of the E3 ubiquitin ligase family (UBE3B). UBE3B is highly expressed immediately after noise in the lesion, but not in the undamaged ends, of the chick basilar papilla. UBE3B is most similar to a ubiquitin ligase gene from Caenorhabditis elegans, suggesting that this gene has been conserved throughout evolution. We also describe preliminary experiments to profile gene expression in the cochlea and brain with commercially available low density gene arrays on nylon membranes and discuss potential applications of this and DNA microarray technology to the auditory system.
Lomax
M I
MI
Kresge Hearing Research Institute, Department of Otolaryngology/Head- Neck Surgery, University of Michigan Medical School, Ann Arbor 48109, USA. mlomax@umich.edu
Huang
L
L
Cho
Y
Y
Gong
T L
TL
Altschuler
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D001489:Basilar Membrane / Q000503:physiopathology; D002645:Chickens; D015870:Gene Expression; D006317:Hearing Loss, Noise-Induced / Q000235:genetics* / Q000503:physiopathology*; D006801:Humans; D007245:Inferior Colliculi / Q000503:physiopathology; D008025:Ligases / Q000235:genetics; D008969:Molecular Sequence Data; D009473:Neuronal Plasticity / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D017386:Sequence Homology, Amino Acid; D044767:Ubiquitin-Protein Ligases
D044767:Ubiquitin-Protein Ligases; D008025:Ligases