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Mol Cell Neurosci. 1993 Feb;4(1):25-9. doi: 10.1006/mcne.1993.1003.

Prolonged Activation of c-fos and Optimal Activation of Pro-opiomelanocortin mRNA after Repeated Morphine Exposure in SH-SY5Y Cells.

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Department of Physiology, Louisiana State University Medical Center, New Orleans, Louisiana 70119; VA Medical Center, New Orleans, Louisiana 70146; and Department of Medicine, Tulane Medical School, New Orleans, Louisiana 70112; Department of Pharmacology, Tulane Medical School, New Orleans, Louisiana 70112; and Alexion Pharmaceutical, New Haven, Connecticut 06511.


The time course of change in the mRNA concentrations of the proto-oncogene c-fos and pro-opiomelanocortin after two different methods of morphine treatment was examined in SH-SY5Y human neuroblastoma cells. In a repeated treatment design, SH-SY5Y cells exposed to morphine sulfate (MS) for 12 h or more were periodically given fresh morphine (10 or 1 muM). In a single-dose design, 10 muM MS was added to the flasks at designated times without changing the medium. Slot-blotting hybridization analysis of total cellular RNA using a [(32)P]-fos cDNA probe revealed that repeated morphine treatment caused both an early transient induction of c-fos and a later prolonged increase in c-fos. Single treatment with morphine caused only a transient and rapid induction of c-fos. Slot-blotting hybridization with a [(32)P]-POMC cRNA probe revealed that POMC mRNA was significantly activated at 6 h and remained significantly elevated up to 7 days in the cells with repeated morphine treatment. In the single-dose experiments, however, the POMC mRNA was not significantly elevated at 2 days or less. It was significantly activated at 6 days, but at a much lower level than that seen in the repeated-dose design. These results indicate that repeated exposure to morphine induces a prolonged activation of c-fos mRNA which may be functionally related to the significant activation of POMC mRNA in SH-SY5Y cells.


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