Responses had to occur during the last 250 ms of the trace period, and the EMG signal had to stay above the predetermined threshold for at least 10 ms for a blink to be classified as a learned response. The learning criterion was set at > 60% learned responses during at least one 100-trial block. When the effects of chemotherapy on retention of trace memories (Fig. 1D) were studied, an even more stringent criterion was used during initial training
– Rats had to express > 60% learned responses during two of three consecutive 100-trial blocks before their ability to remember the conditioned response after administration of TMZ was tested. The highest percentage of learned responses reached MK0683 mw during a 100-trial block was used as an indicator of how well a rat had learned (peak performance). To assess the effects of chemotherapy on hippocampal theta activity, Selleck Ixazomib the relative power of theta activity during a 5-min stimulus-free period immediately preceding the first eyeblink conditioning session (spontaneous) and that induced by the CS during eyeblink conditioning were derived. To examine spontaneous theta activity, the 5-min recording
was divided into 50 artefact-free 3-s sweeps that were used for analysis. To examine induced theta activity, a 500-ms time period starting 250 ms after the onset of the CS was selected for analysis from each conditioning trial, thus avoiding the effect of immediate Branched chain aminotransferase event-related potentials. Sweeps
with artefacts most commonly caused by rapid large-scale movements were automatically rejected from the analysis by simple amplitude thresholding with Matlab. Next, to determine the relative power of hippocampal theta activity [theta/(delta + theta)], a fast Fourier transform was used to analyse the frequency composition of the signal. From the result, the relative power of hippocampal theta activity was determined as the ratio between the power of the signal at 4.5–10.3 Hz and the power of the signal at 1.5–10.3 Hz (theta ratio). Naturally, induced theta ratios were analysed separately for each experiment (Fig. 1B–D). However, regarding the effects of TMZ on spontaneous theta activity, data from two experiments (Fig. 1B and C) were combined to form one group, because the rats in both experiments had been subjected to identical experimental procedures (4 weeks of TMZ/saline) until the first eyeblink conditioning session. Data from the last experiment (Fig. 1D) were used to examine the effects of only 1 week of TMZ/saline treatment on spontaneous theta activity. Rats were euthanised 1 week after the BrdU injection, when the effects of chemotherapy on the retention of a trace memory were assessed (Fig. 1D). In all other experiments (Fig. 1A–C), rats were euthanised 3 weeks after the BrdU injection(s).