期刊
THROMBOSIS RESEARCH
卷 128, 期 5, 页码 E55-E61出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.thromres.2011.06.002
关键词
BDNF; immunoelectron microscopy; thrombin; PAR1; platelet
资金
- Knowledge Cluster Initiative of the Ministry of Education, Sports, Science, and Technology (MEXT)
- Grants-in-Aid for Scientific Research [10J01595] Funding Source: KAKEN
Brain-derived neurotrophic factor (BDNF) is a cytokine that plays important roles in the survival, development, and plasticity of neurons. BDNF is also expressed in peripheral tissues and cells. In this article, we report the BDNF release reaction through thrombin stimulation and its localization in human platelets. Platelets from healthy volunteers were subjected to PAR1-AP or PAR4-AP stimulation. Release of BDNF was measured by ELISA. Localization of BDNF in resting and thrombin-activated platelets was examined by immunoelectron microscopy and sucrose gradient ultracentrifugation following western blotting. BDNF was released dose-dependently with PAR1-AP concentrations with drastic release at low PAR1-AP concentrations and gently release at high PAR1-AP concentrations. Maximum BDNF release was approximately 37% at 132 mu M PAR1-AP. In contrast, 3.8% BDNF was released with 1.13 mM PAR4-AP stimulation. In immunoelectron microscopy and sucrose gradient ultracentrifugation analyses, BDNF was detected not only in alpha-granules but also cytoplasm in of the resting platelets, and it was distributed in the swollen open canalicular system fused to alpha-granules at 1 min and disappeared at 5 min after stimulation by thrombin. However, BDNF in cytoplasm remained throughout platelet activation. In conclusions, we demonstrate that BDNF is released from platelets through predominately PAR1 regulation. Furthermore, we identified two pools of BDNF in the alpha-granules and cytoplasm of human platelets, and only BDNF in alpha-granules is released through platelet activation. (C) 2011 Elsevier Ltd. All rights reserved.
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