Elsevier

Neuroscience

Volume 210, 17 May 2012, Pages 384-392
Neuroscience

Neurodegeneration, Neuroprotection, and Disease-Oriented Neuroscience
Research Paper
Strategies to defeat ketamine-induced neonatal brain injury

https://doi.org/10.1016/j.neuroscience.2012.02.015Get rights and content

Abstract

Studies using animal models have shown that general anesthetics such as ketamine trigger widespread and robust apoptosis in the infant rodent brain. Recent clinical evidence suggests that the use of general anesthetics on young children (at ages equivalent to those used in rodent studies) can promote learning deficits as they mature. Thus, there is a growing need to develop strategies to prevent this injury. In this study, we describe a number of independent approaches to address therapeutic intervention. Postnatal day 7 (P7) rats were injected with vehicle (sterile PBS) or the NMDAR antagonist ketamine (20 mg/kg). After 8 h, we prepared brains for immunohistochemical detection of the pro-apoptotic enzyme activated caspase-3 (AC3). Focusing on the somatosensory cortex, AC3-positive cells were then counted in a non-biased stereological manner. We found AC3 levels were markedly increased in ketamine-treated animals. In one study, microarray analysis of the somatosensory cortex from ketamine-treated P7 pups revealed that expression of activity dependent neuroprotective protein (ADNP) was enhanced. Thus, we injected P7 animals with the ADNP peptide fragment NAPVSIPQ (NAP) 15 min before ketamine administration and found we could dose-dependently reverse the injury. In separate studies, pretreatment of P6 animals with 20 mg/kg vitamin D3 or a nontoxic dose of ketamine (5 mg/kg) also prevented ketamine-induced apoptosis at P7. In contrast, pretreatment of P7 animals with aspirin (30 mg/kg) 15 min before ketamine administration actually increased AC3 counts in some regions. These data show that a number of unique approaches can be taken to address anesthesia-induced neurotoxicity in the infant brain, thus providing MDs with a variety of alternative strategies that enhance therapeutic flexibility.

Highlights

▶There is a growing need to develop strategies to prevent anesthesia-induced injury. ▶We describe a number of independent approaches to address therapeutic intervention. ▶Ketamine induced apoptotic injury in P7 rats. ▶NAP, vitamin D3, and low dose ketamine, all attenuated or prevented this injury. ▶Aspirin enhanced the injury.

Section snippets

Treatment groups

All in vivo procedures used in these studies were approved by the Wake Forest University Animal Care and Use Committee and in accordance with NIH guidelines. All efforts were made to reduce the numbers and suffering of animals used. Animals (Sprague–Dawley) were obtained from Harlan (Charlotte, NC, USA). Pups were maintained in the cage with the mother until the day of the experiment (water and food were available ad libitum). For all studies, at P7, pups were divided into roughly equal

Ketamine enhances or represses expression of multiple genes

We have shown elsewhere (Lema Tomé et al., 2006a, Lema Tomé et al., 2006b, Turner et al., 2010) that wholesale changes in gene expression are to be expected following NMDAR blockade. To monitor these changes in a more efficient and comprehensive manner, we turned to the powerful technique of microarray analysis. We compared gene expression in ketamine- or MK801-treated animals with that of vehicle-treated animals. We chose the time point of 8 h because peak induction of the pro-apoptotic enzyme

Discussion

With respect to anesthesia-induced brain injury in neonates, NIH, the FDA, as well as the International Anesthesiology Research Society (IARS) have urged researchers to not only define the scope of the problem but develop therapeutic strategies that target prevention. Thus, whereas past studies from our laboratory have focused on describing NMDAR blockade-induced injury as well as target mechanisms, more recent work has focused on developing ways to prevent this injury. In this article, we have

Acknowledgments

This work was supported by NIH RO1 NS051632, Wake Forest Intramural Research Support, and Mr and Mrs Tab Williams Family Neuroscience Endowment Fund. Animal handling and tissue processing performed by S.G., C.L., and C.P.T. Stereological cell counting and statistical analysis performed by all authors (except L.M. and J.C.). RNA isolation performed by C.L., and microarray procedure and analysis performed by L.M. and J.C. We are also indebted to Lou Craddock for her assistance with the microarray

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