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Ketamine-Induced Toxicity in Neurons Differentiated from Neural Stem Cells

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Abstract

Ketamine is used as a general anesthetic, and recent data suggest that anesthetics can cause neuronal damage when exposure occurs during development. The precise mechanisms are not completely understood. To evaluate the degree of ketamine-induced neuronal toxicity, neural stem cells were isolated from gestational day 16 rat fetuses. On the eighth day in culture, proliferating neural stem cells were exposed for 24 h to ketamine at 1, 10, 100, and 500 μM. To determine the effect of ketamine on differentiated stem cells, separate cultures of neural stem cells were maintained in transition medium (DIV 6) for 1 day and kept in differentiation medium for another 3 days. Differentiated neural cells were exposed for 24 h to 10 μM ketamine. Markers of cellular proliferation and differentiation, mitochondrial health, cell death/damage, and oxidative damage were monitored to determine: (1) the effects of ketamine on neural stem cell proliferation and neural stem cell differentiation; (2) the nature and degree of ketamine-induced toxicity in proliferating neural stem cells and differentiated neural cells; and (3) to provide information regarding receptor expression and possible mechanisms underlying ketamine toxicity. After ketamine exposure at a clinically relevant concentration (10 μM), neural stem cell proliferation was not significantly affected and oxidative DNA damage was not induced. No significant effect on mitochondrial viability (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay) in neural stem cell cultures (growth medium) was observed at ketamine concentrations up to 500 μM. However, quantitative analysis shows that the number of differentiated neurons was substantially reduced in 10 μM ketamine-exposed cultures in differentiation medium, compared with the controls. No significant changes in the number of GFAP-positive astrocytes and O4-positive oligodendrocytes (in differentiation medium) were detected from ketamine-exposed cultures. The discussion focuses on: (1) the doses and time-course over which ketamine is associated with damage of neural cells; (2) how ketamine directs or signals neural stem cells/neural cells to undergo apoptosis or necrosis; (3) how functional neuronal transmitter receptors affect neurotoxicity induced by ketamine; and (4) advantages of using neural stem cell models to study critical issues related to ketamine anesthesia.

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Abbreviations

CNS:

Central nervous system

l-Ca:

Acetyl-l-carnitine

DMEM:

Dulbecco’s modified Eagle’s medium

ELISA:

Enzyme-linked immunosorbant assay

TUNEL:

Terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labeling

MTT:

3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide

LDH:

Lactate dehydrogenase

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Competing Interests

No external funding and no competing interests declared.

Disclaimer

This document has been reviewed in accordance with United States Food and Drug Administration (FDA) policy and approved for publication. Approval does not signify that the contents necessarily reflect the position or opinions of the FDA. The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the FDA.

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Authors and Affiliations

  1. Division of Neurotoxicology, National Center for Toxicological Research, HFT-132/Food and Drug Administration, Jefferson, AR, 72079, USA

    Fang Liu, Tucker A. Patterson, Natalya Sadovova, Merle G. Paule & Cheng Wang

  2. Department of Anesthesiology, University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA

    Shuo W. Rainosek

  3. Center for Drug Evaluation and Research/Food and Drug Administration, Silver Spring, MD, 20993, USA

    Joseph P. Hanig

  4. Office of the Director, National Center for Toxicological Research/Food and Drug Administration, Jefferson, AR, 72079, USA

    William Slikker Jr.

Authors
  1. William Slikker Jr.
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  2. Fang Liu
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  3. Shuo W. Rainosek
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  4. Tucker A. Patterson
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  5. Natalya Sadovova
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  6. Joseph P. Hanig
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  7. Merle G. Paule
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  8. Cheng Wang
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Correspondence to William Slikker Jr. or Cheng Wang.

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Slikker, W., Liu, F., Rainosek, S.W. et al. Ketamine-Induced Toxicity in Neurons Differentiated from Neural Stem Cells. Mol Neurobiol 52, 959–969 (2015). https://doi.org/10.1007/s12035-015-9248-5

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  • DOI: https://doi.org/10.1007/s12035-015-9248-5

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