Transcription profiling by array of chicken quiescent and proliferating auditory epithelia

ABSTRACT: In order to gain insight into the molecular events which underlie auditory hair cell regeneration in chicken, we compared gene expression in chicken basilar papillae after 24, 48, and 72 hours in culture with or without forskolin (100uM).

Under sterile conditions, cochlear ducts containing the basilar papillae were carefully dissected out of ~4-day-old chicks and then individually cultured in DMEM with 10% fetal bovine serum with or without forskolin (final concentration 100 ?M, delivered in 1% DMSO) for either 24, 48, or 72 hours at 37ï¾°C. Control samples received DMSO at 1% as a vehicle control. At the end of 3 days, the tegmentum vasculosum was dissected off to expose the auditory epithelium, which was delicately freed from the underlying cartilaginous plates. All explants were kept in culture for 3 days because new hair cells are first seen in basilar papillae treated with forskolin after ~3 days of exposure to the drug [19]. Therefore, at even earlier time points in culture the molecular events that subserve hair cell proliferation are well underway. Each sample was comprised of 3 auditory epithelia from 3 different chicks which were put in ~100 ?L of DMEM and then immediately frozen at -80ï¾°C until RNA isolation could be performed. There were a total of 24 samples in this experiment: six 72-hour forskolin, six 72-hour control, three 48-hour forskolin, three 48-hour control, three 24-hour forskolin, and three 24-hour control.

ORGANISM(S): Gallus gallus

SUBMITTER: Corey Frucht 

PROVIDER: E-MEXP-2642 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress