[Abstract Title]. - Society for Neuroscience

Disclosures: B.N. Buran, None; N. Strenzke, None; A. Neef, None; T. Moser, None; M.C.
Liberman, None.
Poster
259. Cochlear Mechanoreceptors
Time: Sunday, November 16, 2008, 1:00 pm - 5:00 pm
Program#/Poster#: 259.9/BB5
Topic: D.02.a. Mechanoreceptors and cochlea
Support: NIH Grant DC05989
Title: Identification and amplification of gap junction genes and proteins from celloidinembedded
human cochlear sections
Authors: X.-H. WANG, *H.-B. ZHAO;
Dept Otolaryngol, Univ. Kentucky Med. Sch., Lexington, KY
Abstract: Gap junctional coupling plays a crucial role in hearing. Connexin mutations are
associated with a high incidence of nonsyndromic hearing loss. However, its pathological
mechanisms remain largely undetermined. In this study, the expressions of gap junction genes
and proteins in the celloidin-embedded human temporal bone sections with formalin fixation
were investigated. After de-celloidin, the genomic DNA was successfully extracted from the
long-term storage human cochlear slice (10 years) using RecoverAll TM Total Nucleic Acid
Isolation kit (Ambion). The formalin-fixed, celloidin-embedded human cochlear sections (10-20
years) were also processed by the antigen-retrieval treatment for immunofluorescent staining.
Connexin 26 (Cx26), which is a predominant isoform of gap junctions in the cochlea, was
identifiable from the PCR product and immunofluorescent staining. Clearly labeling for Cx26
could be detected in the supporting cells in the organ of Corti and at the stria vascularis (SV) and
spiral ligament (SPL) in the cochlear lateral wall. We also examined the expression of Pannexin
(Panx), which is a new found gap junction gene family in vertebrate, in the human cochlea. As
our finding in the rat and mouse cochlea, the Panx1 expression was also detected in the human
cochlea. Our preliminary data show successful extraction of DNA and immunofluorescent
staining in the long-term storage human cochlear sections. These studies provide a
methodological fundament for further identification of gap junction gene expression and
pathogenic mechanisms of deafness mutants in the human cochlea.
Supported by NIH R01 DC05989
Disclosures: X. Wang, None; H. Zhao, None.