Table 2. Variables for induction of IHKA-SE and chronic epilepsy outcomes.
For each animal we note the subject ID, sex, genotype, experimental use, treatment (KA, Saline), dose (nL of KA or Saline), total duration of anesthesia (defined as the time anesthesia started to the time anesthesia ended), latency to the first stage 5 seizure (defined as the time to a stage 5 convulsion after the end of anesthesia). We also note the total number of stage 5 seizures and chronic seizures (2–4 and 10–12 wk seizures were pooled). We also note whether animals were administered with 0.9% of Saline s.c. after IHKA (typically the day after IHKA i.e., 24 h post-IHKA).
| Subject ID | Cage ID | Sex | Genotype | Experimental use | Treatment | Dose (nL) | Total duration of anesthesia (min) | Latency to Stg 5 (min) from IHKA injection | Total number of Stg5 during SE | Total number of chronic seizures | Administration of 0.9% Saline s.c. post-IHKA |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 75a-5 | Male | Cre +/− | EEG, Anatomy | KA | 100 | 17 | 11 | 4 | 168 | No |
| 2 | 76–1 | Female | Cre +/− | EEG, Anatomy | KA | 100 | 17 | 28 | 8 | 8 | No |
| 3 | 76–4 | Female | Cre −/− | EEG, Anatomy | KA | 100 | 15 | 27 | 10 | 136 | Yes |
| 4 | 77–2 | Female | Cre +/− | EEG, Anatomy | KA | 100 | 15 | 42 | 6 | 59 | Yes |
| 5 | 78–3 | Female | Cre −/− | EEG, Anatomy | KA | 70 | 16 | 49 | 5 | 132 | No |
| 6 | 78a-5 | Male | Cre −/− | EEG, Anatomy | KA | 80 | 22 | 44 | 19 | 46 | Yes |
| 7 | 80–4 | Female | Cre −/− | EEG, Anatomy | KA | 70 | 19 | 66 | 4 | 8 | No |
| 8 | 81–1 | Female | Cre −/− | EEG, Anatomy | KA | 70 | 19 | 79 | 2 | 79 | No |
| 9 | 99a-6 | Male | Cre −/− | EEG | KA | 100 | 33 | 60 | 6 | 17 | No |
| 10 | Jack a-1 | Male | NA | EEG, Anatomy | KA | 80 | 21 | 17 | 5 | 44 | No |
| 11 | Jack a-2 | Male | NA | EEG, Anatomy | KA | 90 | 22 | 47 | 9 | 40 | No |
| 12 | CRL a-1 | Male | NA | EEG, Anatomy | KA | 70 | 18 | 30 | 8 | 54 | No |
| 13 | CRL a-2 | Male | NA | EEG, Anatomy | KA | 100 | 20 | 52 | 10 | 15 | No |
| 14 | 78a-6 | Male | Cre +/− | Anatomy | KA | 75 | 20 | 63 | 2 | NA | No |
| 15 | 79a-6 | Male | Cre −/− | Anatomy | KA | 70 | 17 | 39 | 8 | NA | No |
| 16 | 82–1 | Female | Cre +/− | Anatomy | Saline | 100 | 21 | NA | NA | NA | NA |
| 17 | 82–2 | Female | Cre +/− | Anatomy | Saline | 100 | 21 | NA | NA | NA | NA |
| 18 | 82a-3 | Male | Cre −/− | Anatomy | Saline | 80 | 21 | NA | NA | NA | NA |
| 19 | 82a-4 | Male | Cre −/− | Anatomy | Saline | 100 | 19 | NA | NA | NA | NA |
| 20 | 99–1 | Female | Cre −/− | EEG HFOs | Saline | 70 | 27 | NA | NA | NA | NA |
| 21 | 99–3 | Female | Cre +/− | EEG HFOs | Saline | 100 | 29 | NA | NA | NA | NA |
| 22 | 99a-5 | Male | Cre +/− | EEG HFOs | Saline | 70 | 28 | NA | NA | NA | NA |
| 23 | 100–4 | Female | Cre −/− | EEG HFOs | Saline | 100 | 31 | NA | NA | NA | NA |
| 24 | Jax a-3 | Male | NA | EEG HFOs | Saline | 100 | 21 | NA | NA | NA | NA |
| 25 | Jax a-4 | Male | NA | EEG HFOs | Saline | 100 | 22 | NA | NA | NA | NA |
IHKA dose: There was no significant correlation between IHKA dose and the total number of stage 5 seizures during SE (r = 0.19, p = 0.5), suggesting the dose had little influence on the severity of SE.
Anesthesia: There were no significant correlations between the total duration of anesthesia and the latency to the first stage 5 seizure (r = 0.33, p = 0.26), the total number of stage 5 seizures during SE (r = 0.07, p = 0.77), or the total number of chronic seizures (r = −0.47, p = 0.1), suggesting that the duration of anesthesia had little influence on outcome after IHKA.
Latency to the first stage 5 and number of stage 5 seizures during SE: Like the lack of correlation between anesthesia and latency to the first stage 5 seizure, there was no correlation between the latency to the first stage 5 and the total number of stage 5 seizures during SE (r = −0.23, p = 0.39) or total number of chronic seizures (r = −0.38, p = 0.19). These data suggest little effect of the latency of the first stage 5 seizures on the severity of SE or subsequent chronic seizures. The total number of stage 5 seizures during SE was not correlated with the total number of chronic seizures (r = −0.19, p = 0.53) which is surprising because one might expect more severe SE to lead to more severe epilepsy.
Genotype: We did not find any statistically significant differences between animals that were Cre −/− (n = 7) or Cre +/− (n = 4) in terms of the latency to the first stage 5 seizure (Mann-Whitney U test, U = 8, p = 0.31) or the total number of stage 5 seizures during SE (Mann-Whitney U test, U = 10, p = 0.50). Also, the total number of chronic seizures was not different between Cre −/− (n = 6) vs. Cre+/− (n = 3) animals (Mann-Whitney U test, U = 8.5, p = 0.96). These data suggest Cre−/− and Cre+/− mice were similar, regarding the measurements in this table. Also, the C57BL6 background strain was not different from Cre+/+ or Cre+/− mice in terms of the latency to the first stage 5 seizure (Cre+/+: Mann-Whitney U test, U = 7, p = 0.88, Cre+/−: Mann-Whitney U test, U = 8, p = 0.31) or the total number of stage 5 seizures during SE (Cre+/+: Mann-Whitney U test, U = 2.5, p = 0.14, Cre+/−: Mann-Whitney U test, U = 10.5, p = 0.59). Also, the mean number of chronic seizures in the background strain was not different from either Cre+/+ (Mann-Whitney U test, U = 4, p = 0.62) or Cre+/− (Mann-Whitney U test, U = 8, p = 0.47) mice. These data support the view that the use of either Cre+/+ or Cre+/− or the background strain led to no statistically significant effects in acute or chronic epilepsy outcomes examined in this study.
Administration of Saline: Animals that received Saline s.c. the day after IHKA (n = 3) because of transient body weight loss were similar to those that did not (n = 10) in terms of total number of chronic seizures (Mann-Whitney U test, U = 8, p = 0.27).
Orange: IHKA animals; Blue: Saline-injected controls. NA: Not applicable. Jack: Mice from The Jackson Laboratory; CRL: Mice from Charles River Laboratories.