Characterization of dynorphin-containing neurons on dissociated dentate gyrus cell cultures.
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.
Neuropharmacology Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709.
X P
XP
He
P H
PH
Lee
K R
KR
Pennypacker
R K
RK
Tuominen
E C
EC
Mar
L
L
Thai
J S
JS
Hong
D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D002455:Cell Division / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004399:Dynorphins / Q000032:analysis*; D006624:Hippocampus / Q000737:chemistry* / Q000166:cytology; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D009474:Neurons / Q000737:chemistry*; D009479:Neuropeptides / Q000032:analysis; D012333:RNA, Messenger / Q000235:genetics*; D011863:Radioimmunoassay; D051381:Rats; D011916:Rats, Inbred F344
D015415:Biomarkers; D009479:Neuropeptides; D012333:RNA, Messenger; D004399:Dynorphins