Dailey M E, Smith S J
Department of Molecular and Cellular Physiology, Stanford University Medical School, California 94305-5426, USA.
J Neurosci. 1996 May 1;16(9):2983-94. doi: 10.1523/JNEUROSCI.16-09-02983.1996.
Time-lapse fluorescence confocal microscopy was used to directly visualize the formation and dynamics of postsynaptic target structures (i.e., dendritic branches and spines) on pyramidal neurons within developing tissue slices. Within a 2 week period of time, pyramidal neurons in cultured slices derived from early postnatal rat (postnatal days 2-7) developed complex dendritic arbors bearing numerous postsynaptic spines. At early stages (1-2 d in vitro), many fine filopodial protrusions on dendrite shafts rapidly extended (maximum rate approximately 2.5 microM/minute) and retracted (median filopodial lifetime, 10 min), but some filopodia transformed into growth cones and nascent dendrite branches. As dendritic arbors matured, the population of fleeting lateral filopodia was replaced by spine-like structures having a low rate of turnover. This developmental progression involved a transitional stage in which dendrites were dominated by persistent (up to 22 hr) but dynamic spiny protrusions (i.e., protospines) that showed substantial changes in length and shape on a timescale of minutes. These observations reveal a highly dynamic state of postsynaptic target structures that may actively contribute to the formation and plasticity of synaptic connections during CNS development.
采用延时荧光共聚焦显微镜直接观察发育中的组织切片内锥体神经元上突触后靶结构(即树突分支和棘突)的形成和动态变化。在2周的时间内,来自新生大鼠早期(出生后第2 - 7天)的培养切片中的锥体神经元发育出带有许多突触后棘突的复杂树突分支。在早期阶段(体外培养1 - 2天),树突轴上许多细小的丝状伪足迅速伸展(最大速率约为2.5微米/分钟)并回缩(丝状伪足的平均寿命为10分钟),但一些丝状伪足会转变为生长锥和新生树突分支。随着树突分支成熟,短暂的侧向丝状伪足群体被周转速率较低的棘状结构所取代。这种发育进程涉及一个过渡阶段,在此阶段树突由持续存在(长达22小时)但动态变化的棘状突起(即原棘突)主导,这些原棘突在几分钟的时间尺度上长度和形状会发生显著变化。这些观察结果揭示了突触后靶结构的高度动态状态,这可能在中枢神经系统发育过程中积极促进突触连接的形成和可塑性。