Use of the teleost saccule to identify genes involved in inner ear function.
The vertebrate inner ear sensory epithelia contain different types of hair cells and supporting cells. The teleost saccule is anatomically similar to the mammalian saccule and is primarily involved in the detection of translational acceleration and orientation with respect to gravity. To facilitate molecular studies of the teleost saccule cDNA libraries were constructed from microdissected Lepomis macrochirus (bluegill sunfish) saccular maculae. To our knowledge, this is the first report of cDNA libraries constructed from the saccule. In one instance, a non-polymerase chain reaction-based method of amplifying a mRNA population from limited amounts of starting tissue was employed that allowed construction of cDNA libraries from nanogram amounts of tissue mRNA. Conventional cDNA libraries were constructed from the sunfish saccular maculae as well. These cDNA libraries enriched in hair cell and supporting cell transcripts should facilitate molecular biological studies of inner ear sensory epithelia. As an example of their utility, efforts to identify tyrosine kinases expressed in the saccular endorgan using low-stringency hybridization screening of these cDNA libraries and the partial sequence of a cDNA found to encode an erbB-2-related tyrosine kinase are also reported.
Davis
J G
JG
Department of Pathology, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Oberholtzer
J C
JC
Burns
F R
FR
Lee
A M
AM
Saunders
J
J
Eberwine
J H
JH
Greene
M I
MI
D000595:Amino Acid Sequence; D000818:Animals; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics; D004790:Enzyme Induction; D005784:Gene Amplification; D015723:Gene Library; D006198:Hair Cells, Auditory / Q000502:physiology; D008322:Mammals / Q000033:anatomy & histology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010473:Perciformes / Q000033:anatomy & histology / Q000235:genetics*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D012444:Saccule and Utricle / Q000502:physiology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D018076:DNA, Complementary; D020794:Receptor Protein-Tyrosine Kinases; D018719:Erb-b2 Receptor Tyrosine Kinases