Expression of genes implicated in oxidative stress in the cochlea of newborn rats

Oxidative stress is an important mechanism inducing ototoxicity-, age- and noise-induced hearing loss. To better understand this phenomenon, we examined cochlear tissues for the expression of following genes involved directly or indirectly in the oxidative stress response: glyceraldehyde-3-phosphate dehydrogenase (Gapdh); solute carrier family-2 (facilitated glucose transporter), member-1 (Slc2a1); heme oxygenase-1 (Hmox1); heme oxygenase-2 (Hmox2); inducible nitric oxide synthase-2 (Nos2); transferrin (Tf); transferrin receptor (Tfrc); glutathione S-transferase A3 (Gsta3) and metallothionein-1a (Mt1a). Cochlear tissues were dissected from the p3-p5 Wistar rats, divided into the organ of Corti (OC), modiolus (MOD) and stria vascularis together with spiral ligament (SV + SL) and processed immediately or cultured under normoxic conditions or a short-term, mild hypoxia followed by re-oxygenation. After 24 h, explants were collected and total RNA isolated, transcribed and amplified in the real time RT-PCR. We found all genes listed above expressed in the freshly isolated cochlear tissues. In the OC and MOD, Slc2a1, Tf, and Mt1a were expressed on a lower level than in the SV + SL. In the OC, Hmox1 was expressed on a lower level than in the MOD and SV + SL. Hypoxic and normoxic cultures increased the transcript number of Gapdh, Slc2a1 and Hmox1 in all cochlear tissues. The expression of Nos2, Tf, Gsta3 and Mt1a increased in a tissue-specific manner. In the SV + SL, Mt1a expression decreased after normoxic and hypoxic conditions. Taken together, using real time RT-PCR, our results imply that oxidative stress may be an important component of cochlear injury during the developing period. In spite of the immaturity of the tissue, a differential response of antioxidant enzymes/proteins with respect to the pathway, the expression levels and regions was observed.

►Genes implicated in the cochlear oxidative stress. ► Region-specific gene expression in cochlear tissues ex vivo. ► Regulation of gene expression during normoxic and hypoxic culture conditions. ► Region-specific changes in the gene expression after different culture conditions.