Volume 42, Pages 117-128 (January 2015) Urokinase-type plasminogen activator deficiency has little effect on seizure susceptibility and acquired epilepsy phenotype but reduces spontaneous exploration in mice  J. Rantala, S. Kemppainen, X.E. Ndode-Ekane, L. Lahtinen, Tamuna Bolkvadze, K. Gurevicius, H. Tanila, A. Pitkänen  Epilepsy & Behavior  Volume 42, Pages 117-128 (January 2015) DOI: 10.1016/j.yebeh.2014.11.001 Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 1 FFT analysis of cortical EEG in wild-type (Wt, n=8) and uPA−/− (n=8) mice. Percentage of delta, theta, alpha, and beta power during mobility (A) and immobility (B). No differences were observed between the genotypes. The changes in the different frequency bands between the behavioral states were similar between the genotypes. (C) Peak (P) frequency in each frequency band during mobility and immobility. No difference was observed between the genotypes. (D) Also, no differences were found between the genotypes when the 24-hour epochs of the frequency band, up to 80Hz, were analyzed. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 2 (A) A representative sample from a control mouse showing the responses evoked by the first and the second auditory stimulus in the left cortical electrode. Arrows point to responses at 25ms, 50ms, and 180ms after tone onset. (B) Mean amplitudes of the cortical component of auditory-evoked potentials (AEPs) in wild-type (Wt, n=7) and uPA−/− (n=8) mice. Responses to the first and the second stimulus at different latencies (25ms, 50ms, and 180ms) are shown. There were no differences between the phenotypes. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 3 Dot plots summarizing the response of wild-type littermates (Wt, n=7) and uPA−/− mice (n=9) to the administration of pentylenetetrazol (PTZ, 50mg/kg, i.p.). Mice were connected to a video-EEG immediately after PTZ injection and were followed up for 60min. (A) Latency to the first epileptiform spike. (B) Total number of spikes during the 60-minute follow-up. There was no difference between the genotypes. Dashed lines indicate mean value. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 4 Bar graphs summarizing the response of wild-type littermates (Wt, n=6) and uPA−/− mice (n=6) to the administration of pilocarpine (280mg/kg, i.p.). Mice were connected to video-EEG before the administration of pilocarpine and were followed up for up to 60min (or until death). Each bar shows the number of spikes per 10-minute epoch. Note that uPA−/− mice developed spiking more slowly compared with Wt mice, and the spike frequency remained lower than in Wt mice. Data are shown as mean±SEM. Statistical significance: *p<0.05. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 5 Bar graphs summarizing the response of wild-type littermates (Wt, n=7) and uPA−/− mice (n=7) to the administration of intrahippocampal kainate. Mice were connected to video-EEG immediately after kainate injection and were followed for up to 30days (24/7). (A) Analysis of the severity of SE included the first 48h. Each bar shows the number of spikes per 1-hour epoch (no difference between the genotypes). Inset in panel A shows the total number of spikes per 48h in each animal. (B) Latency to the first generalized seizure was not different between the genotypes. (C) Bar graphs summarizing the daily seizure frequency at 1-week epochs in Wt and uPA−/− mice (no difference between the genotypes). (D) Bar graphs summarizing the seizure duration at 1-week epochs in Wt and uPA−/− mice (no difference between the genotypes). Number of seizures included in the analysis is indicated in bars. (E) Daily seizure frequency during the 30-day follow-up. Note the similar cyclicity of seizures in both Wt and uPA−/− mice peaking approximately every 5days (no difference between the genotypes). Data are shown as mean±SEM. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 6 Spontaneous locomotion in wild-type (Wt) and uPA−/− mice. (A) Compared with the Wt mice, the uPA−/− mice showed reduced exploratory activity as indicated by reduced distance traveled during the 10-minute test period [F(1,18)=4.7, p<0.05]. (B) uPA−/− showed reduced rearing time from the first session to the second one compared with Wt mice (session by genotype: F(1,18)=6.1, p<0.05). Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 7 Acoustic startle reaction in wild-type (Wt) and uPA−/− mice. (A) The startle force at different sound intensities was similar between the genotypes (p=0.97). (B) Prepulse administered at 40–60dB attenuated the startle response in uPA mice [F(3,27)=7.5, p=0.001], while Wt mice showed no attenuation [F(3,27)=1.3, p=0.29]. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 8 Fear-conditioning task. (A) uPA−/− mice showed enhanced freezing to a conditioned stimulus (tone) in a fear-conditioning task compared with Wt mice, even before the tone was paired with the unconditioned stimulus (foot shock). The difference was present also on testing day 3 when testing was performed in a new environment (p<0.01). (B) No difference was observed in conditioning to context. Epilepsy & Behavior 2015 42, 117-128DOI: (10.1016/j.yebeh.2014.11.001) Copyright © 2014 Elsevier Inc. Terms and Conditions