Actin bundles in neuronal growth cone observed with the Pol-Scope.
Actin bundles in neuronal growth cone observed with the Pol-Scope.
复制标题
使用 Pol-Scope 观察神经元生长锥中的肌动蛋白束。
DOI：
10.1086/bblv193n2p219
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Oldenbourg,R
中科院分区：
文献类型：
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作者：
Katoh,K;Langford,G;Hammar,K;Smith,PJ;Oldenbourg,R
神经元生长锥出现在树突和轴突的尖端，在树突和轴突生长的导航中起重要作用。生长锥富含肌动蛋白纤维，推测其参与生长锥运动。用荧光显微镜仔细观察标记的肌动蛋白丝，或用视频增强微分干涉对比（DIC）显微镜仔细观察天然肌动蛋白束，揭示了肌动蛋白相关动力学的重要方面，如逆行（1）肌动蛋白的研磨（2），以及肌球蛋白V参与丝状伪足的伸长（3）。但是生长锥运动的分子机制仍然存在争议，因为活细胞中的肌动蛋白束很难可视化。需要新的方法来理解生长锥运动的机制。因此，我们用Pol-Scope（4）观察了活的生长锥中的肌动蛋白束。与传统的偏振光显微镜相比，Pol-Scope显示的双折射元件与其方向无关，并在整个视野内以高灵敏度和分辨率测量其延迟（4）。在本文中，我们证明了使用PollScope观察活体生长锥中未标记的肌动蛋白束的可行性，并将PollScope图像与DIC图像进行了比较。我们根据Kaczmarek和Strumwasser的方法（5）制备袋细胞的原代培养物。培养1 ~ 3天后，细胞形成薄片状的生长锥。
The neuronal growth cone appears at the tip of dendrites and axons where it plays an important role in the navigation of dendritic and axonal growth. Growth cones are rich in actin fibers, which are presumably involved in growth cone movement. Careful observations of labeled actin filaments with a fluorescent microscope or of native actin bundles with a videoenhanced differential interference contrast (DIC) microscope have revealed important aspects of actin-related dynamics, such as retrograde flow (1) treadmilling of actin (2), and the involvement of myosin V in filopodial elongation (3). But the molecular mechanisms of growth cone movement are still in debate, because actin bundles in living cells are difficult to visualize. New methodologies are needed for understanding the mechanisms of growth cone movement. Therefore, we observed actin bundles in living growth cones with the Pol-Scope (4). In contrast to the traditional polarized light microscope, the Pol-Scope reveals birefringent components independent of their orientation, and measures their retardance with high sensitivity and resolution over the whole held of view (4). In this paper, we demonstrate the feasibility of observing unlabeled actin bundles in living growth cones using the Pol-Scope and compare PollScope image with DIC image.Bag cell neurons, from the abdominal ganglion of the marine opisthobranch mollusc, Aplysiu, form relatively large growth cones. We prepared primary cultures of bag cells according to the method of Kaczmarek and Strumwasser (5). The cultured cells formed growth cones shaped like a thin lamellipodium for 1 to 3 days.