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

doi:10.1016/0006-8993(92)91032-A

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

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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 language	English
Pages (from-to)	91-98
Number of pages	8
Journal	Brain Research
Volume	594
Issue number	1
DOIs	https://doi.org/10.1016/0006-8993(92)91032-A
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.

Funding

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.

Funders	Funder number
National Institutes of Health (NIH)
Fogarty International Center	F05TW004205

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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Cite this

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title = "Characterization of dynorphin-containing neurons on dissociated dentate gyrus cell cultures",

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.",

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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.",

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AU - Mar, Eng Chun

AU - Thai, Linda

AU - Hong, Jau Shyong

N1 - 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.

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N2 - 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.

AB - 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.

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

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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