Characterization of dynorphin-containing neurons on dissociated dentate gyrus cell cultures |
| |
Authors: | Xiao-Ping He Paul H. K. Lee Keith R. Pennypacker Raimo K. Tuominen Eng-Chun Mar Linda Thai Jau-Shyong Hong |
| |
Affiliation: | Neuropharmacology Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709. |
| |
Abstract: | In the dentate gyrus, the synthesis of the opioid peptide, dynorphin, is modulated by a variety of stimuli. In order to elucidate the cellular and molecular mechanisms regulating the synthesis of dynorphin in the hippocampus, we have established a routine primary cell culture of dentate granule neurons and identified granule-like neurons by a characteristic marker, dynorphin, in these cultures. Cultures were prepared from 7-day-old rat pups and maintained in medium with 2% fetal bovine serum. These cultures contained approximately 20% neurons and survived for over 4 weeks. After 2 weeks in culture, neurons expressing dynorphin-A and its messenger RNA were detected using immunocytochemistry and in situ hybridization, respectively. In dentate cultures, enkephalin-, cholecystokinin-, neuropeptide Y- and substance P-positive cells were observed in addition to dynorphin-positive cells with immunocytochemistry. The results suggest that dentate gyrus cell cultures provide a valid in vitro model for studying molecular mechanisms regulating prodynorphin gene expression. |
| |
Keywords: | Hippocampus Neuronal cell culture Immunocytochemistry Dentate granule cell Dynorphin In situ hybridization |
本文献已被 ScienceDirect 等数据库收录! |
|