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We counted cytoplasmic TDP43- or FUS/TLS-positive neurons in which diffuse TDP43 or FUS/TLS signal was detected in cytoplasm, in combination with the absence of nuclear immunoreactivity. Data are expressed as mean?��?SD. At first, we tested for normal distribution of samples with SPSS. Then, statistical analysis was performed by using a one-factor ANOVA, followed by a Tukey post hoc comparison. Statistical analyses were performed in SPSS (version 13.0; SPSS Inc., Chicago, IL). All tests were considered statistically significant at P?GUCY1B3 2A, top; n?=?5). There was no change of the density of the CA1 pyramidal neurons between the SC group and the gerbil group subjected to 2-min tCCAO once from 1 day to 6 months (Fig. 1; n?=?5 at each time point; Fig. 2A, left, B, left, open bars). However, in the gerbil group subjected to 2-min tCCAO three times (Fig. 1; n?=?5 at each time point), time-dependent damaged neurons with pycnotic nucleus and neuronal loss were observed in the CA1 region (Fig. 2A, right, B, right, solid bars). Although most hippocampal CA1 neurons survived at 1 day after 3 �� 2-min tCCAO, large numbers of CA1 neurons were gradually lost from 3 to 7 days (2,693.2?��?303.1 cells/mm2 at mTOR inhibitor 3 days, *P?Pexidartinib supplier At 6 months, the CA1 cell density had recovered to 86.5% of SC (Fig. 2). TDP43 immunostaining in the CA1 region is shown in Figure 3. In SC gerbils, TDP43 was weakly stained in the nuclei of some CA1 pyramidal neurons (Fig. 3, top). After a single 2-min tCCAO, the TDP43 immunoreactivity was slowly but progressively induced in CA1 pyramidal neurons, with a peak at 3 months, which slightly returned at 6 months (Figs. 3, left, 5A, left, open bars). From the baseline level of SC (1,396.8?��?130.6 cells/mm2), the total numbers of TDP43-positive cells increased to 1,514.5?��?151.2 cells/mm2 at 1 month (*P?