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Electron microscopy techniques employed to explore mitochondrial defects in the developing rat brain following ketamine treatment. | LitMetric

Electron microscopy techniques employed to explore mitochondrial defects in the developing rat brain following ketamine treatment.

Exp Cell Res

NCTR/ORA Nanotechnology Core Facility, Office of Scientific Coordination, National Center for Toxicological Research, US Food and Drug Administration, 3900 NCTR Road, Jefferson, AR 72079, United States. Electronic address:

Published: December 2018

AI Article Synopsis

  • Ketamine, an FDA-approved anesthetic, has been found to cause significant cell death in developing brains of rats, raising concerns about its safety in pediatric use.
  • This pilot study utilized advanced electron microscopy techniques, namely transmission electron microscopy (TEM) and serial block-face scanning electron microscopy (SBF-SEM), to analyze mitochondrial changes in the brains of rats treated with ketamine.
  • Results indicated that ketamine exposure leads to notable mitochondrial swelling and potential fission in neurons, with three-dimensional analysis revealing larger mitochondrial volumes compared to untreated neurons, highlighting ketamine's neurotoxic effects.

Article Abstract

Ketamine, an FDA-approved N-methyl-D-aspartate (NMDA) receptor antagonist, is commonly used for general pediatric anesthesia. Accumulating evidence has indicated that prolonged exposure to ketamine induces widespread apoptotic cell death in the developing brains of experimental animals. Although mitochondria are known to play a pivotal role in cell death, little is known about the alterations in mitochondrial ultrastructure that occur during ketamine-induced neurotoxicity. The objective of this pilot study was to utilize classic and contemporary methods in electron microscopy to study the impact of ketamine on the structure of mitochondria in the developing rat brain. While transmission electron microscopy (TEM) was employed to comprehensively study mitochondrial inner membrane topology, serial block-face scanning electron microscopy (SBF-SEM) was used as a complementary technique to compare the overall mitochondrial morphology from a representative treated and untreated neuron. In this study, postnatal day 7 (PND-7) Sprague-Dawley rats were treated with ketamine or saline (6 subcutaneous injections × 20 mg/kg or 10 ml/kg, respectively, at 2-h intervals with a 6-h withdrawal period after the last injection, n=6 each group). Samples from the frontal cortex were harvested and analyzed using TEM or SBF-SEM. While classic TEM revealed that repeated ketamine exposure induces significant mitochondrial swelling in neurons, the newer technique of SBF-SEM confirmed the mitochondrial swelling in three dimensions (3D) and showed that ketamine exposure may also induce mitochondrial fission, which was not observable in the two dimensions (2D) of TEM. Furthermore, 3D statistical analysis of these reconstructed mitochondria appeared to show that ketamine-treated mitochondria had significantly larger volumes per unit surface area than mitochondria from the untreated neuron. The ultrastructural mitochondrial alterations demonstrated here by TEM and SBF-SEM support ketamine's proposed mechanism of neurotoxicity in the developing rat brain.

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Source
http://dx.doi.org/10.1016/j.yexcr.2018.10.009DOI Listing

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