Method of euthanasia affects amygdala plasticity in horizontal brain slices from mice

An important consideration in any terminal experiment is the method used for euthanizing animals. Although the prime consideration is that the method is humane, some methods can have a dramatic impact on experimental outcomes. The standard inhalant anesthetic for experiments in brain slices is isoflurane, which replaced the flammable ethers used in the pioneer days of surgery. To our knowledge, there are no data available evaluating the effects of the method of euthanasia on plasticity changes in brain slices. Here, we compare the magnitude of long-term potentiation (LTP) and long-term depression (LTD) in the lateral nucleus of the amygdala (LA) after euthanasia following either ether or isoflurane anesthesia, as well as in mice decapitated without anesthesia. We found no differences in input–output curves using different methods of euthanasia. The LTP magnitude did not differ between ether and normal isoflurane anesthesia. After deep isoflurane anesthesia LTP induced by high frequency stimulation of cortical or intranuclear afferents was significantly reduced compared to ether anesthesia. In contrast to ether anesthesia and decapitation without anesthesia, the low frequency stimulation of cortical afferents induced a reliable LA-LTD after deep isoflurane anesthesia. Low frequency stimulation of intranuclear afferents only caused LTD after pretreatment with ether anesthesia. The results demonstrate that the method of euthanasia can influence brain plasticity for hours at least in the interface chamber. Therefore, the method of euthanasia is an important consideration when brain plasticity will be evaluated.

► We examine changes in amygdala plasticity that are dependent on the type of anesthesia administered prior to decapitation and brain slice preparation.
► After decapitation without anesthesia and after deep isoflurane anesthesia LTP was significantly reduced compared to ether anesthesia.
► Low frequency stimulation (LFS) of intranuclear afferents only caused LTD after pretreatment with ether anesthesia.
► LFS of cortical afferents only induced a reliable LA-LTD after deep isoflurane anesthesia.
► The results show that method of euthanasia is an important consideration when brain plasticity will be evaluated in brain slices.