解剖学报

解剖学报 ›› 2016, Vol. 47 ›› Issue (4): 456-461.doi: 10.16098/j.issn.0529-1356.2016.04.004

• 神经生物学 • 上一篇    下一篇

甘油二酯激酶γ参与大鼠胚胎脑神经上皮细胞的发育

牛宝贝1 崔慧林1* 曹锡梅1 师亮1 马晋峰2 乔从进1   

  1. 1. 山西医科大学组织学胚胎学教研室,太原 030001; 2. 山西医科大学附属肿瘤医院普外科，太原 030013
  • 收稿日期:2016-03-10 修回日期:2016-04-17 出版日期:2016-08-06 发布日期:2016-08-06
  • 通讯作者: 崔慧林 E-mail:cuihuilin99@163.com
  • 基金资助:

    山西省优势重点学科项目；山西医科大学基础医学院331基础医学科技培植基金计划

Diacylglycerol kinase γ is involved in the neuroepithelium development of rat embryonic brain

NIU Bao-bei1 CUI Hui-lin 1* CAO Xi-mei1 SHI Liang1 MA Jin-feng2 QIAO Cong-jin1   

  1. 1.Department of Histology and Embryology，Shanxi Medical University，Taiyuan 030001，China; 2.Department o f General Surgery， the Tumor Hospital Affiliated to Shanxi Medical University，Taiyuan 030013，China
  • Received:2016-03-10 Revised:2016-04-17 Online:2016-08-06 Published:2016-08-06
  • Contact: CUI Hui-lin E-mail:cuihuilin99@163.com

PDF

摘要：

目的 探讨甘油二酯激酶γ(DGKγ)在大鼠胚胎脑神经上皮发育中的作用及可能机制。 方法 18只胎龄（E）11.5、E12.5、E14.5、E16.5、E18.5大鼠胚胎用于Western blotting方法检测DGKγ在大鼠胚胎脑组织的表达，E9.5～E18.5大鼠胚胎各5只用于免疫组织化学和免疫荧光染色方法检测DGKγ在大鼠胚胎脑的分布以及DGKγ与Ki67、乙酰胆碱转移酶（ChAT）及酪氨酸羟化酶（TH）的细胞内共定位。 结果 DGKγ蛋白含量在E11.5～E14.5表达逐渐增多，E16.5表达较E14.5显著减少，E18.5表达显著增多且高于E14.5水平。E10.5～E12.5，DGKγ在脑泡壁出现表达，并随着脑泡的发育表达在端脑、除大脑脚外的中脑、后脑和末脑；E13.5～E14.5，DGKγ的强阳性表达从海马及周围的新皮质、苍白球、嗅脑和大脑脚阳性延伸到除新皮质外的脑的各区域；E15.5至出生前，DGKγ在新皮质阳性表达增强，但在大脑脚和脑桥表达减弱。免疫荧光染色显示，E14.5，与海马和苍白球相比，嗅脑的DGKγ阳性细胞中Ki67阳性率较高（87%），ChAT 阳性率较低（9%）（均P＜0.05），中脑顶盖的DGKγ阳性细胞中62%为TH阳性；E16.5，新皮质的DGKγ阳性细胞中Ki67阳性率为26%，较E14.5时（49%）显著降低（P＜0.05），ChAT阳性率为70%，较E14.5时（45%）显著增高（P＜0.05）；DGKγ多位于神经上皮细胞的胞质，也可见于胞膜上，或者同时位于胞质和胞核。结论 DGKγ在胚胎脑的表达模式和亚细胞分布，提示其参与了胚胎脑神经上皮的增殖、迁移和分化后神经元的发育。

关键词: 甘油二酯激酶γ, 神经上皮, 胚胎脑, 免疫印迹法, 免疫组织化学, 免疫荧光, 大鼠

Abstract:

Objective To investigate the effects and possible mechanisms of diacylglyceral kinase γ (DGKγ) in the neuroepithelium development of the rat embryonic brain.Methods Western blotting was performed to examine the expression of DGKγ in 18 rat embryonic brains at embryonic day(E)11.5, E12.5, E14.5, E16.5 and E18.5. Five rat embryos each day from E9.5 to E18.5 were stained by immunohistochemistry and immunofluorescence to detect the distribution of DGKγ in rat embryonic brains and the colocalization of DGKγ with Ki67/choline acetyltramsferase(ChAT)/tyrosine hydroxylase(TH) in neuroepithelial cells. Results The expression of DGKγ protein increased gradually from E11.5 to E14.5, decreased significantly at E16.5 than that at E14.5, and the expression of DGKγ at E18.5 increased obviously to the level and higher than that at E14.5. At E10.5-E12.5, DGKγ was expressed from one brain vesicle to telencephalon, mesencephalon, rhombencephalon and myelencephalon except the tegmentum. At E13.5-E14.5, DGKγ was strongly expressed in hippocampus and its adjacent neocortex, pallidum, rhinencephalon and tegmentum, and then extending to all regions of brain except the neocortex. From E15.5 to before birth, the expression of DGKγ increased in neocortex and weakened in tegmentum and pons. Immunofluorescence staining showed that in DGKγ positive cells of rhinencephalon, Ki67 positive rate was higher (87%) and ChAT positive rate was lower (9%) than that of hippocampus and pallidum (all P<0.05). In tectum of E14.5 rat, 62% DGKγ positive cells showed TH positive reaction. In DGKγ positive cells of neocortex, Ki67 positive rate significantly decreased from 49% (E14.5) to 26% (E16.5), while ChAT positive rate was significantly increased from 45% (E14.5) to 70% (E16.5) (all P<0.05). The expression of DGKγ was observed in cytoplasm of neuroepithelial cells, or in membrane, or in both nucleus and cytoplasm. Conclusion The expression pattern of DGKγ in the rat embryonic brain and the subcellular distribution of DGKγ suggest that DGKγ is involved in the proliferation, migration and development of neurons.

Key words: Diacylglyceral kinase γ, Neuroepithelium, Embryonic brain, Western blotting, Immunohistochemistry, Immunofluorescence, Rat