Cisplatin induces cytoplasmic to nuclear translocation of nucleotide excision repair factors among spiral ganglion neurons

Preincision complex-I from the excision nuclease reaction among cochlear spiral limbus and outer hair cells

“…The sections were then pretreated with a blocking solution of normal horse or goat serum, 10% Triton X-100 and 2% bovine serum albumin (BSA; Sigma, St. Louis, MO, USA) in PBS for 1 h. The primary antibodies were diluted in the blocking solution at a 1:200 concentration. The primary antibodies anti-XPC and anti-XPA (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA) have been characterized previously through preabsorption experiments, immunoreactivity and Western blots (Guthrie, in press;Guthrie et al 2008;Wu et al 2007a;Romanenko et al 2002;Jaitovich-Groisman et al 2001;Yang et al 2008). For negative controls the primary antibody was omitted Yang et al 2008).…”

“…The expression of several excision nuclease genes including XPC and XPA have been quantified from the kidney using quantitative RT-PCR (Cheng et al 1999;Guthrie et al 2008). Additionally, excision nuclease rates in the rat kidney have been shown to be higher than other tissues such as brain, heart, muscle, lung and spleen, (Gospodinov et al 2003).…”

“…Such adducts have been localized among the various cochlear cell-types and recent experiments have shown an increase in cochlear XPA and XPC gene expression and immunoreactivity among spiral ganglion neurons, the stria vascularis and spiral ligament as a function of systemic cisplatin (2 mg/kg/day, i.p.) treatment (Guthrie in press;Guthrie et al 2008;Thomas et al 2006;van Ruijven et al 2005). However, human cochleae harvested from patients treated with cisplatin reveal that cochlear OHC are among the most vulnerable to cell death while cochlear spiral limbus cells (SLCs) are among the least vulnerable (Cheng et al 2001;Hoistad et al 1998;Strauss et al 1983).…”

Noise Stress Induces an Epidermal Growth Factor Receptor/Xeroderma Pigmentosum–A Response in the Auditory Nerve

“…The primary antibodies are commercially available and included goat anti-EGFR (sc-31155) and rabbit anti-XPA (sc-853) (Santa Cruz Biotechnology, Inc.; Dallas, TX). We and others have previously characterized the validity of these antibodies in preabsorption experiments, immunofluorescent assays, IHC, and Western blots, and the results are as follows 15,21 : (1) Western blots revealed that the EGFR antibody selectively detects the 170-kDa protein product and the XPA antibody selectively detects the 40-kDa protein product, (2) incubating the antibody with the immunogen yields no immunoreactivity in tissue sections, and (3) negative control sections where the antibody is omitted from the IHC procedure do not result in immunoreactivity. Chromogen consisted of 3,3′-diaminobenzidine tetrahydrochloride (DAB) to produce a brown reaction product or DAB+1% cobalt (Co) to produce a blue reaction product.…”

“…For instance, ototoxic exposures such as noise exposure and chemical intoxications result in DNA damage within spiral ganglion neurons. 21,24 In addition to disrupting the functions of active genes, DNA damage is a potent inducer of cell death. 25 Therefore, future therapies designed to rescue spiral ganglion functions after ototoxic exposure could target XPA activity by manipulating EGFR signaling.…”