Cellular neurochemical characterization and subcellular localization of phospholipase C β1 in rat brain
Cellular neurochemical characterization and subcellular localization of phospholipase C β1 in rat brain
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DOI：
10.1016/j.neuroscience.2012.06.039
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发表时间：
2012-10
期刊：
影响因子：
3.3
通讯作者：
Mario Montaña;Gontzal García del Caño;M. L. D. Jesús;Imanol González-Burguera;L. Echeazarra;S. Barrondo;J. Sallés
中科院分区：
文献类型：
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作者：
Mario Montaña;Gontzal García del Caño;M. L. D. Jesús;Imanol González-Burguera;L. Echeazarra;S. Barrondo;J. Sallés
本研究描述了磷脂酶C β 1（PLCβ1）亚型沿着在成年大鼠神经轴上的完整和详细的神经解剖分布图，并通过双重免疫荧光染色和共聚焦激光扫描确定了表达PLCβ1的细胞的表型，沿着其在皮层神经元中的亚细胞定位。免疫组织化学标记显示，皮质PLCβ1阳性细胞之间存在相当大的形态异质性，尽管锥体细胞形态明显占优势。作为GFAP和PLCβ1分布不匹配的一个例外，在脊髓白色物质的放射状胶质样突起中观察到高度的共表达。在躯体感觉皮层中，与PLCβ1共染色的GABA能神经元的比例在I、II-III、IV和VI层中相似（约2/3），而在V层中显著较低（约2/5）。在大鼠皮层分离的核中，针对PLCβ1和核斑点标记物SC-35和NeuN/Fox 3的双重免疫荧光显示，核基质中这两种标记物与PLCβ1高度重叠。与此相反，有没有明显的共定位标记的核包膜和椎板。最后，为了评估PLCβ1的亚细胞表达模式是否特别涉及PLCβ1的两种剪接变体之一，我们在皮质亚细胞组分中进行了Western印迹实验。值得注意的是，PLCβ1a/1b比值在细胞质中比在核膜和质膜组分中更高。这些结果提供了PLCβ1亚型在大鼠脑不同细胞亚型中的细胞分布及其在神经元核区室中的存在的更深入的知识。
The present study describes a complete and detailed neuroanatomical distribution map of the phospholipase C beta1 (PLCβ1) isoform along the adult rat neuraxis, and defines the phenotype of cells expressing PLCβ1, along with its subcellular localization in cortical neurons as assessed by double-immunofluorescence staining and confocal laser scanning. Immunohistochemical labeling revealed a considerable morphological heterogeneity among PLCβ1-positive cells in the cortex, even though there was a marked predominance of pyramidal morphologies. As an exception to the general non-matching distribution of GFAP and PLCβ1, a high degree of co-expression was observed in radial glia-like processes of the spinal cord white matter. In the somatosensory cortex, the proportion of GABAergic neurons co-stained with PLCβ1 was similar (around 2/3) in layers I, II–III, IV and VI, and considerably lower in layer V (around 2/5). Double immunofluorescence against PLCβ1 and nuclear speckle markers SC-35 and NeuN/Fox3 in isolated nuclei from the rat cortex showed a high overlap of both markers with PLCβ1 within the nuclear matrix. In contrast, there was no apparent co-localization with markers of the nuclear envelope and lamina. Finally, to assess whether the subcellular expression pattern of PLCβ1 involved specifically one of the two splice variants of PLCβ1, we carried out Western blot experiments in cortical subcellular fractions. Notably, PLCβ1a/1b ratios were statistically higher in the cytoplasm than in the nuclear and plasma membrane fractions. These results provide a deeper knowledge of the cellular distribution of the PLCβ1 isoform in different cell subtypes of the rat brain, and of its presence in the neuronal nuclear compartment.