Versatile and Simple Approach to Determine Astrocyte Territories in Mouse Neocortex and Hippocampus
Versatile and Simple Approach to Determine Astrocyte Territories in Mouse Neocortex and Hippocampus
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DOI：
10.1371/journal.pone.0069143
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发表时间：
2013-07-23
期刊：
影响因子：
3.7
通讯作者：
Reichenbach, Andreas
中科院分区：
文献类型：
--
作者：
Grosche, Antje;Grosche, Jens;Reichenbach, Andreas
背景：星形胶质细胞除了支持神经元的功能外，还是神经元信息处理的积极伙伴。典型的领土结构的星形胶质细胞（体积的神经元占据的单个星形胶质细胞）是关键的胶质细胞-神经元interactions.Methods的许多方面：个别星形胶质细胞领土体积测量高尔基体浸渍，和星形胶质细胞密度测定S100 β免疫标记。这些数据进行比较，从传统应用的方法，如染料填充和测定生物胞素负载的星形胶质细胞网络的密度的结果。最后，我们实施了我们的新方法，以调查年龄相关的变化，星形胶质细胞领土在皮质和海马的5-和21个月大的mice.Results：我们的简化方法的基础上高尔基体浸渍获得的数据进行了比较，以前发表的染料填充实验，并产生了显着可比的结果，星形胶质细胞领土体积。此外，我们发现，几乎所有的耦合星形胶质细胞（如生物胞素负载所示）的S100 β免疫阳性。这种新的实验方法的第一个应用提供了洞察星形胶质细胞领土体积的年龄依赖性变化。它们随着年龄的增长而增加，而细胞密度保持稳定。在5个月大的小鼠中，重叠因子接近1，显示星形胶质细胞区域很少或没有交错。然而，在21个月大的小鼠，重叠因子超过2，这表明相邻的星形胶质细胞interdigitate.Conclusion的过程：在这里，我们验证了一个简单的，通用的方法来评估星形胶质细胞的领土和相邻领土之间的重叠因子的可用性。其次，我们发现星形胶质细胞的领土体积与年龄相关，导致非重叠领土中的严格组织丧失。未来的研究应该阐明这种适应性反应的星形胶质细胞在衰老的大脑和本研究中提出的方法可能是一个强大的工具，这样做的生理相关性。
Background: Besides their neuronal support functions, astrocytes are active partners in neuronal information processing. The typical territorial structure of astrocytes (the volume of neuropil occupied by a single astrocyte) is pivotal for many aspects of glia-neuron interactions.Methods: Individual astrocyte territorial volumes are measured by Golgi impregnation, and astrocyte densities are determined by S100 beta immunolabeling. These data are compared with results from conventionally applied methods such as dye filling and determination of the density of astrocyte networks by biocytin loading. Finally, we implemented our new approach to investigate age-related changes in astrocyte territories in the cortex and hippocampus of 5- and 21-month-old mice.Results: The data obtained by our simplified approach based on Golgi impregnation were compared to previously published dye filling experiments, and yielded remarkably comparable results regarding astrocyte territorial volumes. Moreover, we found that almost all coupled astrocytes (as indicated by biocytin loading) were immunopositive for S100 beta. A first application of this new experimental approach gives insight in age-dependent changes in astrocyte territorial volumes. They increased with age, while cell densities remained stable. In 5-month-old mice, the overlap factor was close to 1, revealing little or no interdigitation of astrocyte territories. However, in 21-month-old mice, the overlap factor was more than 2, suggesting that processes of adjacent astrocytes interdigitate.Conclusion: Here we verified the usability of a simple, versatile method for assessing astrocyte territories and the overlap factor between adjacent territories. Second, we found that there is an age-related increase in territorial volumes of astrocytes that leads to loss of the strict organization in non-overlapping territories. Future studies should elucidate the physiological relevance of this adaptive reaction of astrocytes in the aging brain and the methods presented in this study might be a powerful tool to do so.