Identification of genes expressed after noise exposure in the chick basilar papilla.
We used differential display of mRNA, a method based on reverse transcriptase-PCR, to identify genes whose expression increases in response to acoustic trauma in the chick basilar papilla. Identifying these genes would provide insight into processes involved in repair of the damaged epithelium or in hair cell regeneration. We compared mRNA from the basilar papilla of normal chicks, from chicks exposed to an octave band noise (center frequency: 1.5 kHz) presented at 118 dB for 6 h, and from chicks exposed to noise and allowed to recover for 2 days. Thus far, we have identified 70 bands that appear to be differentially displayed on DNA sequencing gels; approximately 40 of these bands have been subcloned and sequenced. DNA sequences were compared with sequences in the GenBank database to identify genes with significant (70-85%) sequence identity to known genes. Chick cDNAs identified included: the parathyroid hormone-related protein, an immediate early gene; the delta-subunit of the neuronal-specific Ca2+/calmodulin-regulated protein kinase II; and the GTP-binding protein CDC42, a member of the ras superfamily of G proteins. A fourth cDNA had 84% sequence identity to an uncharacterized human cDNA (expressed sequence tag), indicating that this is a novel gene. Slot-blot hybridization analysis of these cDNAs probed with labeled DNA generated from mRNA from each experimental group indicated higher levels of mRNA for each of these four genes after noise exposure. These results indicate the potential involvement of both Ca2+/calmodulin-mediated signaling and GTPase cascades in the response to noise damage and during hair cell regeneration in the chick basilar papilla.
Gong
T W
TW
Kresge Hearing Research Institute, Department of Otolaryngology/Head-Neck Surgery, University of Michigan Medical School, Ann Arbor 48109-0648, USA.
Hegeman
A D
AD
Shin
J J
JJ
Adler
H J
HJ
Raphael
Y
Y
Lomax
M I
MI
D000161:Acoustic Stimulation; D000818:Animals; D001483:Base Sequence; D001489:Basilar Membrane / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry* / Q000235:genetics; D004848:Epithelium / Q000293:injuries; D019204:GTP-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics* / Q000502:physiology; D011905:Genes, ras / Q000235:genetics; D006198:Hair Cells, Auditory / Q000166:cytology / Q000473:pathology; D008969:Molecular Sequence Data; D009622:Noise / Q000009:adverse effects; D009693:Nucleic Acid Hybridization; D010281:Parathyroid Hormone / Q000235:genetics / Q000378:metabolism; D044162:Parathyroid Hormone-Related Protein; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D010281:Parathyroid Hormone; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins; D012333:RNA, Messenger; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019204:GTP-Binding Proteins