Characterization of dynorphin-containing neurons on dissociated dentate gyrus cell cultures

Xiao Ping He, Paul H.K. Lee, Keith R. Pennypacker, Raimo K. Tuominen, Eng Chun Mar, Linda Thai, Jau Shyong Hong

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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 languageEnglish
Pages (from-to)91-98
Number of pages8
JournalBrain Research
Volume594
Issue number1
DOIs
StatePublished - 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

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