Davis L, Burger B, Banker G A, Steward O
Department of Neuroscience, University of Virginia, School of Medicine, Charlottesville 22908.
J Neurosci. 1990 Sep;10(9):3056-68. doi: 10.1523/JNEUROSCI.10-09-03056.1990.
We have previously reported that recently synthesized RNA is selectively transported into the dendrites of hippocampal neurons grown in culture (Davis et al., 1987). The present study provides further details about this transport process, focusing especially on the velocity of transport, by comparing the velocity of dendritic transport of RNA in neurons of different ages and in the branched and unbranched dendrites of individual neurons. In our previous study, we recognized that calculations of transport velocity could be compromised because transport was being evaluated in a population of dendrites of varying lengths. The present study uses a mathematical modeling approach to determine how the morphology of the population of dendrites would affect the analysis of transport velocity. Focusing first on a simple model, we compared the distribution of transported material at various times when all dendrites were of the same length and when the population included dendrites of different lengths. We found that the distance of labeling increased linearly over time when all dendrites were of the same length, but increased with a negatively accelerating curve when dendrites were of different lengths. We then determined the actual distribution of dendritic lengths in cultured hippocampal neurons, based on immunostaining with an antibody directed against the selective dendritic marker, microtubule-associated protein 2 (MAP2). Using a computer model, we calculated the mean distance of transport as a function of time in this population of dendrites, assuming different velocities of transport. The velocity that best fit the measured distances of RNA transport in both 7- and 15-d-old neurons was 11 microns/hr (0.26 mm/d). However, for the dendrites exhibiting the longest distance of labeling, the best-fitting curve assumed a velocity of 21 microns/hr in both 7- and 15-d-old neurons (0.50 mm/d). Comparisons of transport in branched and unbranched dendrites revealed that the distance of labeling over branched dendrites was consistently longer than over unbranched dendrites of individual neurons. However, neurons with a larger proportion of branched dendrites did not exhibit a greater mean distance of transport. The density of silver grains was higher over branched than over unbranched dendrites, suggesting that a greater amount of recently synthesized RNA may be transported into branched dendrites. Taken together, these results suggest that RNA transport into dendrites is regulated differentially in the dendrites of individual neurons.
我们之前曾报道,最近合成的RNA会被选择性地转运到培养的海马神经元的树突中(戴维斯等人,1987年)。本研究提供了关于这一转运过程的更多细节,特别是通过比较不同年龄神经元以及单个神经元的分支和未分支树突中RNA的树突转运速度,重点关注转运速度。在我们之前的研究中,我们认识到转运速度的计算可能会受到影响,因为转运是在不同长度的树突群体中进行评估的。本研究采用数学建模方法来确定树突群体的形态如何影响转运速度的分析。首先关注一个简单模型,我们比较了所有树突长度相同时以及群体中包含不同长度树突时不同时间点转运物质的分布。我们发现,当所有树突长度相同时,标记距离随时间呈线性增加,但当树突长度不同时,标记距离随负加速曲线增加。然后,我们基于用针对选择性树突标记物微管相关蛋白2(MAP2)的抗体进行免疫染色,确定了培养的海马神经元中树突长度的实际分布。使用计算机模型,我们计算了在假设不同转运速度的情况下,该树突群体中转运平均距离随时间的函数关系。最符合7日龄和15日龄神经元中RNA转运测量距离的速度为11微米/小时(0.26毫米/天)。然而,对于标记距离最长的树突,最拟合曲线在7日龄和15日龄神经元中均假设速度为21微米/小时(0.50毫米/天)。对分支和未分支树突中的转运进行比较发现,分支树突上的标记距离始终长于单个神经元未分支树突上的标记距离。然而,分支树突比例较大的神经元并未表现出更大的平均转运距离。分支树突上的银颗粒密度高于未分支树突,这表明可能有更多最近合成的RNA被转运到分支树突中。综上所述,这些结果表明RNA向树突的转运在单个神经元的树突中受到不同的调节。