Development and Aging of Neuronal Synapse

(A)树突棘细胞骨架蛋白的分析我们使用与抗树突棘单克隆抗体M2 F6结合的珠制备树突棘中的细胞骨架蛋白复合物。所制备的样品含有肌动蛋白，肌球蛋白I，肌球蛋白II，肌球蛋白V和凝溶胶蛋白除了dreglutamine本身。此外，该样品中还有许多其他未鉴定的蛋白质;因此，我们使用该样品作为免疫原制备了单克隆抗体。因此，我们提出了抗肌球蛋白抗体和抗CaMK Ⅱ抗体。抗肌球蛋白抗体特异性识别非肌肉肌球蛋白II重链，其除了树突棘之外还存在于细胞索马和树突干中。这表明棘细胞因子的特征是肌动蛋白结合蛋白，而不是肌球蛋白。(B)脊髓细胞骨架在发育和衰老过程中的变化。在发育中的脑中，在迁移的神经元和生长的轴突和树突中观察到drexine E。在成年人的大脑中，dreploid A位于树突中， 关于我们 IC棘。在这项研究中，我们调查了在发育过程中，树突状细胞何时从细胞索马体中消失并在树突棘中富集。用日本MBL公司生产的抗dreplastin单克隆抗体对大脑和小脑进行免疫组化染色，结果表明，dreplastin在出生后0 ~ 10天的细胞索马和树突中呈免疫反应性，尽管dreplastin的表达总体上随出生后天数的增加而减少。而14日龄大鼠的神经元中仅观察到点状的dreximab免疫反应。树突索马胞体和树突内几乎未观察到树突免疫反应。这些数据表明，dreplant亚细胞定位在出生后12天左右同步变化，表明神经元细胞骨架的最终成熟可能发生在出生后12天。应用100 μM谷氨酸可减弱树突棘上的drexin免疫反应性。转染实验表明，dreplastin A在神经元中的过表达导致脊髓长度的延长。这些数据表明，树突状细胞从树突轴到树突棘的分布的发育变化可能与突触功能的成熟有关。少

(A) Analysis of spine cytoskeletonsWe prepare the cytoskeletal protein complex in the dendritic spine using the bead bound to anti-drebrin monoclonal antibody M2F6. The prepared sample contained actin, myosin I, myosin II, myosin V and gelsolin in addition to drebrin itself. Further, there are many other unidentified proteins in that sample ; therefore, we raised monoclonal antibodies using this sample as an immunogen. As a consequence we raised anti-myosin antibody and anti-CaMKII antibody. The anti-myosin antibody specifically recognized non-muscle myosin II heavy chan, which is present in cell soma and dendritic shaft in addition to dendritic spine. This indicates that spine cytoskelton is characterized by actin binding proteins but not by myosins.(B) The change of spine cytoskeletons during development and aging.In the developing brain, drebrin E is observed in the migrating neurons and within the growing axons and dendrite. In the adult brain, drebrin A is localized in the dendrit … More ic spines. In this study we investigated when drebrin disappeared from cell soma and enriched in dendritic spines during development. Immunohistochemical staining of the cerebrum and cerebellum using anti-drebrin monoclonal antibody (MBL, Japan) demonstrated that drebrin immunoreactivity was observed in the cell soma and dendrites from postnatal 0 day to 10 day, although drebrin expression as a whole was decreased according to the postnatal days. However, in 14-day-old rat, drebrin immunoreactivity was only observed as dot-like pattern in neuropil. Drebrin immunoreactivity was hardly observed within the cell soma and dendrites. These data indicates that drebrin subcellular localization was changed synchronously around postnatal 12 days, indicating that the final maturation of neuronal cytoskeleton may occurred at postnatal 12 days. Application of 100 μM glutamate weakened drebrin immunoreactivity at dendritic spines. Transfection experiments demonstrated that overexpression of drebrin A in neuron resulted the elongation of the spine length. These data indicate that developmental changes in the distribution of drebrin from dendritic shafts into dendritic spines may be related to the maturation of synaptic function. Less