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Morphine does not affect astrocyte survival in developing primary mixed-glial cultures
1Department of Anatomy and Neurobiology, University of Kentucky School of Medicine, Lexington 40536-0084.
Brain Research. Developmental Brain Research
|December 17, 1993
Summary
High concentrations of morphine do not kill glial cells. Instead, morphine reduces glial cell numbers by decreasing the proliferation rate of astrocytes, not by increasing cell death.
Area of Science:
- Neuroscience
- Cell Biology
Background:
- High morphine concentrations (1 microM) previously inhibited glial proliferation but not DNA synthesis in astrocytes.
- This suggested potential morphine toxicity to glial cells.
Purpose of the Study:
- To assess morphine toxicity in mixed-glial cultures.
- To determine if morphine increases glial cell death.
Main Methods:
- Mixed-glial cultures were treated with morphine.
- Cell viability was assessed using calcein-AM (live) and ethidium homodimer (dead) dyes.
- Immunocytochemistry for glial fibrillary acidic protein (GFAP) and A2B5 was combined with viability markers.
Main Results:
- No significant difference in dead cell numbers between morphine-treated and untreated groups at 3, 5, and 7 days in vitro.
- Cell death rates decreased over time in vitro (16-19% at 3 days, 8-12% at 7 days).
- Only GFAP-positive and A2B5-positive cells underwent cell death; flat, polyhedral GFAP-positive cells did not die.
Conclusions:
- Morphine does not increase glial cell death.
- Morphine-induced reductions in glial numbers are due to decreased proliferation, not increased toxicity.
- Culture conditions may influence cell survival more than morphine treatment.