目的研究宫内暴露尼古丁对生后小鼠大脑皮层神经细胞凋亡的影响。方法建立妊娠期宫内暴露尼古丁小鼠动物模型,观察生后21天小鼠大脑皮层组织结构变化,免疫组织化学方法和W estern b lot方法检测宫内暴露尼古丁模型大脑皮层caspase-3的...目的研究宫内暴露尼古丁对生后小鼠大脑皮层神经细胞凋亡的影响。方法建立妊娠期宫内暴露尼古丁小鼠动物模型,观察生后21天小鼠大脑皮层组织结构变化,免疫组织化学方法和W estern b lot方法检测宫内暴露尼古丁模型大脑皮层caspase-3的表达。结果尼古丁组小鼠大脑皮层脑回厚度较对照组变薄(P<0.05),免疫组织化学法检测到大脑皮层caspase-3阳性反应物的平均光密度值在尼古丁组高于对照组(P<0.05),W estern b lot方法检测到大脑皮层caspase-3酶原(32KD)形式及活性片段形式(17KD、20KD)的相对表达量高于对照组(P<0.01)。结论孕期宫内暴露尼古丁激活生后小鼠大脑皮层神经细胞caspase-3的过表达,参与大脑皮层神经细胞凋亡。展开更多
BACKGROUND： Biological and morphological characteristics of neural stem/progenitor cells （NSPCs） have been widely investigated. OBJECTIVE： To explore the ultrastructure of human embryo-derived NSPCs and neurospher...BACKGROUND： Biological and morphological characteristics of neural stem/progenitor cells （NSPCs） have been widely investigated. OBJECTIVE： To explore the ultrastructure of human embryo-derived NSPCs and neurospheres cultivated in vitro using electron microscopy. DESIGN, TIME AND SETTING： A cell biology experiment was performed at the Brain Tumor Laboratory of Soochow University, and Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University between August 2007 and April 2008. MATERIALS： Human fetal brain tissue was obtained from an 8-week-old aborted fetus; serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium was provided by Gibco, USA; scanning electron microscope was provided by Hitachi Instruments, Japan; transmission electron microscope was provided by JEOL, Japan. METHODS： NSPCs were isolated from human fetal brain tissue and cultivated in serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium. Cells were passaged every 5-7 days. After three passages, NSPCs were harvested and used for ultrastructural examination. MAIN OUTCOME MEASURES： Ultrastructural examination of human NSPCs and adjacent cells in neurospheres. RESULTS： Individual NSPCs were visible as spherical morphologies with rough surfaces under scanning electron microscope. Generally, they had large nuclei and little cytoplasm. Nuclei were frequently globular with large amounts of euchromatin and a small quantity of heterochromatin, and most NSPCs had only one nucleolus. The Golgi apparatus and endoplasmic reticulum were underdeveloped; however, autophagosomes were clearly visible. The neurospheres were made up of NSPCs and non-fixiform material inside. Between adjacent cells and at the cytoplasmic surface of apposed plasma membranes, there were vesicle-like structures. Some membrane boundaries with high permeabilities were observed between some contiguous NSPCs in neurospheres, possibly attributable to plasmalemmal fusion between adjacent cells. CONCLUSION： A large number of autophagosomes were observed in NSPCs and gap junctions were visible between adjacent NSPCs.展开更多
文摘目的研究宫内暴露尼古丁对生后小鼠大脑皮层神经细胞凋亡的影响。方法建立妊娠期宫内暴露尼古丁小鼠动物模型,观察生后21天小鼠大脑皮层组织结构变化,免疫组织化学方法和W estern b lot方法检测宫内暴露尼古丁模型大脑皮层caspase-3的表达。结果尼古丁组小鼠大脑皮层脑回厚度较对照组变薄(P<0.05),免疫组织化学法检测到大脑皮层caspase-3阳性反应物的平均光密度值在尼古丁组高于对照组(P<0.05),W estern b lot方法检测到大脑皮层caspase-3酶原(32KD)形式及活性片段形式(17KD、20KD)的相对表达量高于对照组(P<0.01)。结论孕期宫内暴露尼古丁激活生后小鼠大脑皮层神经细胞caspase-3的过表达,参与大脑皮层神经细胞凋亡。
基金the National Natural Science Foundation of China,No.30400457the National Natural Science Foundation of China,No.30672164+1 种基金the National Natural Science Foundation of China,No.30772241the Natural Science Foundation of Jiangsu Province,China, No.BK2007507
文摘BACKGROUND： Biological and morphological characteristics of neural stem/progenitor cells （NSPCs） have been widely investigated. OBJECTIVE： To explore the ultrastructure of human embryo-derived NSPCs and neurospheres cultivated in vitro using electron microscopy. DESIGN, TIME AND SETTING： A cell biology experiment was performed at the Brain Tumor Laboratory of Soochow University, and Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University between August 2007 and April 2008. MATERIALS： Human fetal brain tissue was obtained from an 8-week-old aborted fetus; serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium was provided by Gibco, USA; scanning electron microscope was provided by Hitachi Instruments, Japan; transmission electron microscope was provided by JEOL, Japan. METHODS： NSPCs were isolated from human fetal brain tissue and cultivated in serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium. Cells were passaged every 5-7 days. After three passages, NSPCs were harvested and used for ultrastructural examination. MAIN OUTCOME MEASURES： Ultrastructural examination of human NSPCs and adjacent cells in neurospheres. RESULTS： Individual NSPCs were visible as spherical morphologies with rough surfaces under scanning electron microscope. Generally, they had large nuclei and little cytoplasm. Nuclei were frequently globular with large amounts of euchromatin and a small quantity of heterochromatin, and most NSPCs had only one nucleolus. The Golgi apparatus and endoplasmic reticulum were underdeveloped; however, autophagosomes were clearly visible. The neurospheres were made up of NSPCs and non-fixiform material inside. Between adjacent cells and at the cytoplasmic surface of apposed plasma membranes, there were vesicle-like structures. Some membrane boundaries with high permeabilities were observed between some contiguous NSPCs in neurospheres, possibly attributable to plasmalemmal fusion between adjacent cells. CONCLUSION： A large number of autophagosomes were observed in NSPCs and gap junctions were visible between adjacent NSPCs.
