Figure 1.
Expressions of ER stress-related molecules GRP78 and CHOP in the mouse hippocampus with or without tunicamycin treatment.
C57BL/6J mice were intracerebroventricular injected with tunicamycin (0.02–1 mg/ml) at various time course (6–72 h). In A, the levels of GRP78 and CHOP proteins in hippocampus were determined by Western blotting 24 h after tunicamycin injection in a dose-dependent manner. Representative images of three independent experiments are shown. In B, the levels of GRP78 and CHOP proteins from hippocampus were determined by Western blotting after tunicamycin (0.1 mg/ml) injection in a dose-dependent manner. In C, the levels of GRP78 and CHOP mRNAs from hippocampus were determined by real-time PCR after tunicamycin (0.1 mg/ml) injection in a time-dependent manner. In B and C, quantification of proteins and mRNA levels were shown. Data are presented as mean ± S.E.M. (n = 3). *P<0.05 as compared with sham-control.
Figure 2.
Chop gene and protein expressions in the hippocampus isolated from wild type and chop knockout mice.
Chop genome typing and protein expression in the hippocampus isolated from C57BL/6J mice and chop−/− mice 1 day after intracerebroventricular injection of tunicamycin (0.1 and 1 mg/ml) were detected by the PCR (A) and Western blotting (B), respectively. Representative images of three independent experiments are shown.
Figure 3.
Expressions of ER stress-related molecules GRP78, IRE-1, phospho-IRE-1, XBP-1, JNK, phospho-JNK, phospho-PERK, phospho-eIF2α, and ATF6 in the hippocampus isolated from wild type and chop knockout mice.
The expressions of GRP78, IRE-1, phospho-IRE-1, XBP-1, JNK, phospho-JNK, phospho-PERK, phospho-eIF2α, and ATF6 in the hippocampus isolated from C57BL/6J and chop−/− mice 5 days after intracerebroventricular injection of tunicamycin were performed by the Western blotting (A). Quantification of protein expressions were shown in B. Data are presented as means ± S.E.M. for three independent experiments. Each data was performed in triplicate. *P<0.05 as Tun+Wild Type versus Tun+chop−/− mice.
Figure 4.
Expressions of pro-caspases 9 and 12 in the hippocampus isolated from wild type and chop−/− mice.
Tunicamycin (0.1 mg/ml) were intracerebroventricular injected in the hippocampus of C57BL/6J and CHOP−/− mouse for 3–5 days. Pro-caspase 12 and pro-caspase 9 expressions were presented by the Western blotting from day 3 to day 5 after tunicamycin injection. Representative images of three independent experiments are shown.
Figure 5.
Expressions of pro-caspases/caspases 9 and 12 in the hippocampus isolated from wild type and chop−/− mice.
(A) Tunicamycin (0.1 mg/ml) was intracerebroventricular injected in the hippocampus of C57BL/6J and CHOP−/− mouse for 3–5 days. Pro-caspase 12 and pro-caspase 9 expressions were presented by the Western blotting from day 3 to day 5 after tunicamycin injection. Representative images of three independent experiments are shown. (B) The expressions of pro-caspase 12, pro-caspase 9, and cleaved caspase 9 in the hippocampus of 57BL/6J and chop−/− mice at day 5 after treatment with tunicamycin (0.1 mg/ml) were detected by the Western blotting. Quantification of pro-caspase 12 (B-a) and cleaved caspase 9 (B-b) were shown. Data are presented as mean ± S.E.M. for each group (n = 3). *P<0.05 as Tun+Wild Type versus Tun+chop−/− mice.
Figure 6.
Cell apoptosis in the hippocampus or cortex regions were isolated from wild type and chop−/− mice.
Tunicamycin (0.1 mg/ml) were injected in the hippocampus of wild type C57BL/6J and CHOP−/− mouse. Apoptotic cells were performed by the TUNEL staining 5 days after tunicamycin injection. The staining of neuron nuclei in normal control hippocampus was shown on top (A), and the TUNEL positive cells from hippocampus or cortex (B) of wild type C57BL/6J and chop−/− mouse were quantified (C). Data are presented as mean ± S.E.M. for each group (n = 4). **P<0.01 as Tun+wild type versus Tun+chop−/− mice. H: hippocampus; C: cortex.
Figure 7.
Neuronal living cells in hippocampus of wild type and CHOP−/− mice.
Tunicamycin (0.1 mg/ml) were intracerebroventricular injected in the hippocampus of wild type C57BL/6J (W; A) and CHOP−/− (B) mouse for 5–7 days. Neuronal cells were identified by immunofluorescence staining with Neu-N (green fluorescence). Representative images of three independent experiments are shown. The star symbol indicates that the neuronal living cells are markedly decreased in hippocampus of CHOP−/− mice.
Figure 8.
Behavior analysis in wild type and CHOP−/− mice with or without tunicamycin treatment.
(A). Passive avoidance was performed 1 day before and 5 days after administration of tunicamycin (0.1 mg/ml) in wild type and CHOP−/− mice. The avoidance latency is significantly decreased in chop−/− mice than in wild type mice 5 days after tunicamycin administration. (B). Water maze was performed 1 day before and 5 days after administration of tunicamycin (0.1 mg/ml) in wild type and chop−/− mice. The time spent in finding the target platform is significantly increased in chop −/− mice than in wild type mice 5 days after tunicamycin administration. (C). In water maze, the time spent in the target quadrant is significantly decreased in chop−/− mice than in wild type mice 5 days after tunicamycin (0.1 mg/ml) administration. Data are presented as mean ± S.E.M. for each group (n = 5). *P<0.05 as compared with wild type mice or chop−/− mice without tunicamycin.