Brain capillaries and cholinergic neurons persist in organotypic brain slices in the absence of blood flow
Brain capillaries and cholinergic neurons persist in organotypic brain slices in the absence of blood flow
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DOI：
10.1046/j.1460-9568.2003.02728.x
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发表时间：
2003-07-01
影响因子：
3.4
通讯作者：
Humpel, C
中科院分区：
文献类型：
--
作者：
Moser, KV;Schmidt-Kastner, R;Humpel, C
血管生成在大脑发育过程中起着重要作用，在病理条件下也是如此。本研究的目的是观察器官型脑片中脑毛细血管和胆碱能神经元之间的相互作用。免疫组织化学方法显示脑毛细血管样结构(RecA-1抗原)和胆碱能神经元(胆碱乙酰转移酶)。在正常培养条件下，2周龄和4周龄的大脑皮层脑片中的毛细血管数量非常少。用酸性介质(PH 6)或高温(42℃)处理脑片可显著增加脑毛细血管的数量。与10 ng/mL血管内皮生长因子孵育只能促进更发达的切片中的血管生成。胆碱能神经元在Meynert基底核的切片中存活，高温而不是酸中毒使其数量显著减少。在Meynert基底核和皮质(用酸性介质处理)的切片中，大量的RecA-1阳性毛细血管和胆碱能神经元持续存在，并显示出大量的神经纤维生长到皮质内。总之，我们已经证明，在没有血液灌流的情况下，可以在切片培养中研究RecA-1阳性的毛细血管和胆碱能神经元，该模型可以为研究与血管性痴呆有关的机制提供一个系统。
Angiogenesis plays an important role during development of the brain and under pathological conditions. The aim of the present study was to observe interaction of brain capillaries and cholinergic neurons in organotypic brain slices. Immunohistochemistry was used to visualize brain capillary-like structures (RECA-1 antigen) and cholinergic neurons (choline acetyltransferase). Under normal culture conditions, a very low number of brain capillaries was found in 2- and 4-week-old cortex brain slices. Treatment of slices with acidic medium (pH 6) or hyperthermia (42 degreesC) markedly enhanced the number of brain capillaries. Incubation with 10 ng/mL vascular endothelial growth factor only enhanced angiogenesis in more developed slices. Cholinergic neurons survived in slices of the basal nucleus of Meynert; however, hyperthermia but not acidosis markedly decreased their number. In coslices of the basal nucleus of Meynert and cortex (pretreated with acidic medium), a high number of RECA-1-positive capillaries and cholinergic neurons persisted and displayed strong nerve fibre growth of cholinergic fibres into the cortex. In conclusion, we have demonstrated that RECA-1-positive capillaries and cholinergic neurons can be studied in slice cultures in the absence of blood perfusion, and that this model could provide a system to study mechanisms involved in vascular dementia.