发育中的大鼠脑中神经元特异性二酰基甘油激酶β和γ的免疫细胞化学定位

Naoko Adachi, Miho Oyasu, Taizo Taniguchi, Yasuto Yamaguchi, Rika Takenaka, Yasuhito Shirai, Naoaki Saito
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引用次数: 36

摘要

二酰基甘油激酶(DGK)磷酸化二酰基甘油(DG)产生磷脂酸(PA),因此是DG信号的潜在终止物。DG和PA是细胞内重要的第二信使。DG直接结合蛋白激酶C (PKC)并激活该多功能酶。Ca2+依赖性和脑特异性DGKs, α, β和γ,被认为在中枢神经系统中起关键作用。为了阐明DGK在神经元发育中的作用,我们研究了DGKα、β和γ在出生后大鼠脑中的发育变化。免疫印迹分析发现,DGKα和γ亚型在出生后逐渐升高,而DGKβ亚型在出生后第3天不存在,从第14天开始迅速升高,在第28天达到最大值。免疫组化结果显示,DGKβ和γ分布于不同脑区。在大多数脑区,DGKγ在出生后发育期间持续表达。有趣的是,DGKγ在第3天至第14天在内侧展状核中短暂表达,而DGKγ在浦肯野细胞中表达延迟,这与浦肯野细胞的树突状生长一致。在海马锥体细胞中,DGKβ和γ均丰富,但亚细胞定位不同。DGKγ定位于细胞质中,DGKβ定位于细胞膜结构。这些结果表明,DGK亚型在发育神经元中具有不同的时空功能。
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Immunocytochemical localization of a neuron-specific diacylglycerol kinase β and γ in the developing rat brain

Diacylglycerol kinase (DGK) phosphorylates diacylglycerol (DG) to produce phosphatidic acid (PA) and is, therefore, a potential terminator of DG signaling. DG and PA are important intracellular second messengers. DG directly binds protein kinase C (PKC) then activates this multifunctional enzyme. Ca2+-dependent and brain-specific DGKs, α, β, and γ, are suggested to play pivotal roles in the central nervous system. To elucidate the DGK function in neuronal development, we studied the developmental changes of DGKα, β, and γ in the postnatal rat brain. By immunoblot analysis, DGKα and γ subtypes were present at birth and then gradually increased, while DGKβ was not present at birth or postnatal day 3, then increased rapidly from day 14 to reach maximum at day 28. Immunohistochemically, DGKβ and γ were distributed in different brain regions. In most brain regions, DGKγ showed sustained expression throughout the postnatal developmental periods. Interestingly, a temporal expression of DGKγ was observed in the medial geniculate nucleus during day 3 to 14, and a delay of DGKγ expression was seen in Purkinje cells, which was coincident with dendritic growth of Purkinje cells. In the hippocampal pyramidal cell, both DGKβ and γ were abundant but subcellular localization was different. DGKγ localized in the cytosol while DGKβ localized along the membrane structure. These findings suggest that each DGK subtype has a spatio-temporally different function in the developmental neurons.

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