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.
Original language | English |
---|---|
Pages (from-to) | 91-98 |
Number of pages | 8 |
Journal | Brain Research |
Volume | 594 |
Issue number | 1 |
DOIs | |
State | Published - Oct 23 1992 |
Bibliographical note
Funding Information:Acknowledgements. We would like to acknowledge the excellent technical assistance of Drs. L. Tian and W. Zhang. We also thank Mrs. Loretta Moore for excellent editorial assistance. X.-P.H. is a recipient of a Fogarty International Fellowship (1 FO5 TWO4205-01 BI-5) from the Fogarty International Center of the National Institutes of Health.
Keywords
- Dentate granule cell
- Dynorphin
- Hippocampus
- Immunocytochemistry
- In situ hybridization
- Neuronal cell culture
ASJC Scopus subject areas
- General Neuroscience
- Molecular Biology
- Clinical Neurology
- Developmental Biology