Measurement of the concentrations of dopamine and its metabolites in PC12 cells and in extracellular fluid by detection of native fluorescence

  • Kobayashi Yoko
    Department of Nutrition, Faculty of Health Care, Kiryu University

Bibliographic Information

Other Title
  • 自然蛍光検出法を用いたラット副腎褐色細胞腫(PC12)細胞から放出されるドーパミン量の測定

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Description

Rat pheochromocytoma (PC12) cells synthesize and release neurotransmitters such as dopamine and noradrenaline. Dopamine, which is synthesized from L-tyrosine in cells, is metabolized to dihydroxyphenylacetic acid (DOPAC), 3-methoxytyramine (3MT), and homovanillic acid (HVA). Dopamine is known to show native fluorescence. In this study, we aimed to establish a method to measure the concentrations of dopamine and its metabolites in PC12 cells by using high performance liquid chromatography coupled with a fluorescence detector (HPLC-FL).  First, several concentration of monoamine standard solutions, including that of dopamine, serotonin, and their metabolites, were separated and detected by HPLC-FL. The concentrations of dopamine, 3MT, and HVA were measured with a fluorescence detector set at an excitation wavelength of 225 nm and emission wavelength of 310 nm. However, the sensitivity of DOPAC detection was low. n-Hexanal (hexanal), a straight aldehyde with a 6-carbon chain length, is known to stimulate dopamine release from PC12 cells. Therefore, the concentrations of dopamine and its metabolites in samples from PC12 cells stimulated by hexanal were measured using HPLC-FL. We observed that PC12 cells released dopamine and that the concentrations of extracellular dopamine and 3MT were elevated after stimulation with hexanal. In future, we will attempt to elucidate the physiological significance of the dopamine released by hexanal by using this method.

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Details 詳細情報について

  • CRID
    1390285300182390144
  • NII Article ID
    130007887545
  • DOI
    10.34506/bku.24.0_111
  • ISSN
    24357049
    21864748
  • Text Lang
    ja
  • Data Source
    • JaLC
    • CiNii Articles
  • Abstract License Flag
    Disallowed

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