Expression of prostaglandin E2 synthases in mouse postnatal cortical neurons.
Expression of prostaglandin E2 synthases in mouse postnatal cortical neurons.
复制标题
小鼠出生后皮质神经元中前列腺素 E2 合酶的表达。
DOI：
10.1111/j.1749-6632.2005.tb00056.x
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发表时间：
2005
影响因子：
5.2
通讯作者：
Doré,Sylvain
中科院分区：
文献类型：
--
作者：
Echeverria,Valentina;Greenberg,DavidL;Doré,Sylvain
摘要：类二十烷酸及其代谢相关的酶在神经炎症过程中发挥着积极作用，而神经炎症过程通常是神经退行性疾病的标志。大脑皮层神经元构成神经系统疾病中高度受影响的细胞群。为了获得细胞模型来分析皮质神经元中前列腺素的作用和代谢，我们在无血清培养基中开发了小鼠皮质的产后神经元培养物。培养的皮层细胞在神经元中高度富集，仅含有 5 ± 2% 的神经胶质细胞。检测培养物中几种神经元成熟蛋白标记物的表达：突触标记物（突触蛋白 I 和突触素）和谷氨酸受体亚基（NMDA 受体 1 和谷氨酸受体 1）。还研究了类二十烷酸代谢酶的蛋白质表达，包括环氧合酶-2、微粒体和胞质 PGE2 合酶。出生后神经元在体外无血清培养基中成功存活了长期（长达 40 天），其特征是突触蛋白 I、突触素和微管相关蛋白 2 的表达。谷氨酸受体亚基表达在培养物中随着时间的推移而增加，在 15 天时达到最高水平。参与类二十烷酸代谢的酶遵循独特的表达模式，表明在基础条件下培养物中 PGE2 合成随时间的潜在调节。使用出生后脑培养物具有多种优点，特别是在神经元成熟程度方面，使用出生后幼崽代替怀孕小鼠，并可能增加神经炎症过程和前列腺素细胞神经生物学模型的临床相关性。
Abstract:Eicosanoids and the enzymes associated with their metabolism play an active role in the neuroinflammatory process that is often a hallmark of neurodegenerative disorders. Cerebral cortical neurons constitute a highly affected cell population in neurologic disorders. To obtain a cellular model to analyze prostaglandin action and metabolism in cortical neurons, we developed postnatal neuronal cultures from mouse cortex in a serum‐free medium. Cultured cortical cells were highly enriched in neurons containing only 5 ± 2% glial cells. The cultures were assayed for expression of several protein markers of neuronal maturity: synaptic markers (synapsin I and synaptophysin) and glutamate receptor subunits (NMDA receptor 1 and glutamate receptor 1). The protein expression of eicosanoid‐metabolizing enzymes, including cyclooxygenase‐2 and microsomal and cytosolic PGE2synthases, was investigated as well. Postnatal neurons successfully survived for a long term (up to 40 days)in vitroin serum‐free media, as characterized by the expression of synapsin I, synaptophysin, and microtubule‐associated protein 2. Glutamate receptor subunit expression increased over time in cultures, with the highest levels at 15 days. Enzymes involved in the eicosanoid metabolism followed a distinct pattern of expression, suggesting potential regulation of PGE2synthesis with time in cultures under basal conditions. Use of postnatal brain cultures offers several advantages, especially regarding degree of neuronal maturation, use of postnatal pups instead of pregnant mice, and potentially increased clinical relevance in models of neuroinflammation processes and prostanoid cellular neurobiology.