Zhuxia Zhang1, Jie Yang1, Xiaolei Liu1, Xiaoe Jia1, Shengdi Xu1, Kerui Gong2, Shaochun Yan1, Chunyang Zhang3, Guo Shao4. 1. Biomedicine Research Center and Basic Medical Department, Baotou Medical College, Inner Mongolia, People's Republic of China. 2. Biomedicine Research Center and Basic Medical Department, Baotou Medical College, Inner Mongolia, People's Republic of China; Department of Oral and Maxillofacial Surgery, University of California San Francsico, San Francisco, USA. 3. Department of Neurosurgery, The First Affiliated Hospital of Baotou Medical College, Inner Mongolia, People's Republic of China. Electronic address: zhangchunyangbyyfy@163.com. 4. Biomedicine Research Center and Basic Medical Department, Baotou Medical College, Inner Mongolia, People's Republic of China; Beijing key laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, People's Republic of China. Electronic address: shao_guo_china@163.com.
Abstract
BACKGROUND: Learning and memory is a complex process. Some reports have shown that protein kinases (PKs) and phosphatases (PPs) are important mediators in this process. And it is also well known that protein serine/threonine phosphatase 1 (PP1) and DNA methylation are critically involved in learning and memory. METHODS: In the current study, the mice and cultured cells (NG108-15) were treated with vehicle or 5-Aza-2'-deoxycytidine (5-aza-cdR), a DNA methyltransferase (DNMT) inhibitor. The ability of learning and memory of mice was detected by Morris Water Maze, while the mRNA and protein expression levels of DNMTs and PP1γ in mice hippocampus were measured by real-time PCR and western-blot. To further clarify whether the 5-aza-cdR effects on learning and memory depend on cell proliferation/apoptosis or not, the effects of 5-aza-cdR on the cell proliferation, apoptosis, and PP1γ transcriptional activity were analyzed by using the xCelligence system, flow cytometer and Luciferase reporter assay, respectively. RESULTS: The ability of learning and memory was increased while the expressions of DNMTs and PP1γ were decreased in the hippocampus of mice which were injected with 5-aza-cdR. In vitro experiments showed 10μM 5-aza-cdR inhibited cell proliferation, decreased PP1γ transcription without inducing apoptosis. CONCLUSION: Our data demonstrate that the 5-aza-cdR restrains the expression of PP1γ which is related to learning and memory in the mice.
BACKGROUND: Learning and memory is a complex process. Some reports have shown that protein kinases (PKs) and phosphatases (PPs) are important mediators in this process. And it is also well known that protein serine/threonine phosphatase 1 (PP1) and DNA methylation are critically involved in learning and memory. METHODS: In the current study, the mice and cultured cells (NG108-15) were treated with vehicle or 5-Aza-2'-deoxycytidine (5-aza-cdR), a DNA methyltransferase (DNMT) inhibitor. The ability of learning and memory of mice was detected by Morris Water Maze, while the mRNA and protein expression levels of DNMTs and PP1γ in mice hippocampus were measured by real-time PCR and western-blot. To further clarify whether the 5-aza-cdR effects on learning and memory depend on cell proliferation/apoptosis or not, the effects of 5-aza-cdR on the cell proliferation, apoptosis, and PP1γ transcriptional activity were analyzed by using the xCelligence system, flow cytometer and Luciferase reporter assay, respectively. RESULTS: The ability of learning and memory was increased while the expressions of DNMTs and PP1γ were decreased in the hippocampus of mice which were injected with 5-aza-cdR. In vitro experiments showed 10μM 5-aza-cdR inhibited cell proliferation, decreased PP1γ transcription without inducing apoptosis. CONCLUSION: Our data demonstrate that the 5-aza-cdR restrains the expression of PP1γ which is related to learning and memory in the mice.