Hierarchical Cluster Analysis of Three-Dimensional Reconstructions of Unbiased Sampled Microglia Shows not Continuous Morphological Changes from Stage 1 to 2 after Multiple Dengue Infections in Callithrix penicillata
Hierarchical Cluster Analysis of Three-Dimensional Reconstructions of Unbiased Sampled Microglia Shows not Continuous Morphological Changes from Stage 1 to 2 after Multiple Dengue Infections in Callithrix penicillata
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DOI：
10.3389/fnana.2016.00023
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发表时间：
2016-03-22
影响因子：
2.9
通讯作者：
Picanco Diniz, Cristovam W.
中科院分区：
文献类型：
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作者：
Diniz, Daniel G.;Silva, Geane O.;Picanco Diniz, Cristovam W.
众所周知，小胶质细胞的形态和功能是相关的，但很少有研究探讨小胶质细胞形态变化响应特定病原体的微妙之处。在本报告中，我们定量了病毒侵袭中枢神经系统的登革热病猴模型中小胶质细胞的形态变化。为了模拟流行地区通常发生的多重感染，那里的登革热感染发病率较高，且携带不同血清型的蚊子媒介并存，受试者每周皮下注射一次 DENV3（基因型 III）感染的培养物上清液，24 小时后注射抗 DENV2 抗体。对照动物每周接受抗 DENV2 抗体注射，或不接受注射。对脑切片进行 DENV3 抗原和 IBA-1 免疫标记。从齿状回的多态性层中采集随机和系统的小胶质细胞样本进行 3D 重建，我们发现 INFut 和 DENV3 病毒抗原的强烈免疫染色。我们将小胶质细胞的所有双模态或多模态形态参数提交给层次聚类分析，发现了两种主要的形态表型，分别命名为 I 型和 II 型。与 I 型（第一阶段）相比，II 型小胶质细胞更加复杂；显示更多数量的节点、进程和树以及更大的表面积和体积（第 2 阶段）。 II 型小胶质细胞仅在受感染的猴子中发现，而 I 型小胶质细胞在对照和受感染的受试者中均发现。对对照和 ADE 登革热感染猴子中随机和系统选择的样本进行 3D 重建的形态学参数的层次聚类分析表明，从第 1 阶段到第 2 阶段的小胶质细胞形态变化可能不是连续的。
It is known that microglial morphology and function are related, but few studies have explored the subtleties of microglial morphological changes in response to specific pathogens. In the present report we quantitated microglia morphological changes in a monkey model of dengue disease with virus CNS invasion. To mimic multiple infections that usually occur in endemic areas, where higher dengue infection incidence and abundant mosquito vectors carrying different serotypes coexist, subjects received once a week subcutaneous injections of DENV3 (genotype III)-infected culture supernatant followed 24 h later by an injection of anti-DENV2 antibody. Control animals received either weekly anti-DENV2 antibodies, or no injections. Brain sections were immunolabeled for DENV3 antigens and IBA-1. Random and systematic microglial samples were taken from the polymorphic layer of dentate gyrus for 3-D reconstructions, where we found intense immunostaining for INFut and DENV3 virus antigens. We submitted all bi- or multimodal morphological parameters of microglia to hierarchical cluster analysis and found two major morphological phenotypes designated types I and II. Compared to type I (stage 1), type II microglia were more complex; displaying higher number of nodes, processes and trees and larger surface area and volumes (stage 2). Type II microglia were found only in infected monkeys, whereas type I microglia was found in both control and infected subjects. Hierarchical cluster analysis of morphological parameters of 3-D reconstructions of random and systematic selected samples in control and ADE dengue infected monkeys suggests that microglia morphological changes from stage 1 to stage 2 may not be continuous.