Neurons growing from neonatal murine olfactory explant culture responded to forskolin.
CONCLUSIONS
Olfactory epithelium explant culture represents a model that will be useful in studying the calcium signal transduction of olfactory sensory neurons (OSNs).


OBJECTIVE
The aim of this study was to test the feasibility and efficacy of olfactory epithelium explant culture in the study of calcium signal transduction.


METHODS
Using explant culture, devoid of any enzyme use, a mixed cell type culture system from newborn murine olfactory epithelium was achieved. Morphological and immunochemical criteria were employed to identify the cultured cells. Calcium imaging, using the fluorescent calcium indicator fura-2, was used to monitor changes in the intracellular calcium concentration of candidate OSNs following exposure to forskolin, a cAMP pathway stimulant.


RESULTS
The putative OSNs synthesized both neuronal proteins and olfactory-specific markers. The cultured neurons originated from the 'precursors' present in the olfactory epithelium explants, became fully developed, and could survive for at least 9 days in vitro. Upon stimulation, the intracellular calcium concentration of the OSNs increased from 58.5+/-12.8 nM (n=20) to 577.2+/-57.6 nM (n=10).
Zhang
Luo
L
Key Laboratory Otolaryngology Head and Neck Surgery, Ministry of Education of China, Beijing Institute of Otorhinolaryngology. luozhang@trhos.com
Xian
Mu
M
Fan
Erzhong
E
Wang
Hong
H
Han
Demin
D