脑红蛋白和缺氧诱导因子-1α在牦牛后脑的表达与定位研究

doi:10.11843/j.issn.0366-6964.2021.03.023

摘要/Abstract

摘要： 脑红蛋白（neuroglobin，NGB）和缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）是存在于脊椎动物神经系统中的两种关键性神经保护因子，在低氧适应过程中具有重要的生理功能。为探究牦牛（Bos grunniens）脑组织中对机体运动、呼吸、感觉等生理活动起重要调节作用的后脑在适应低氧过程中NGB和HIF-1α的表达与分布关系，本研究利用RT-qPCR、Western blot及免疫组织化学技术，对NGB和HIF-1α在牦牛后脑不同区域的表达与定位特征进行了研究。RT-qPCR和Western blot结果显示，牦牛后脑中，NGB和HIF-1α基因与蛋白表达趋势基本一致，在小脑蚓前叶中的表达量最高，并显著高于延髓、脑桥及小脑其他部位（P<0.05），小脑半球皮质次之（P<0.05），蚓小叶表达量最低，其他不同区域间两种因子的表达量也存在一定差异。免疫组化结果显示，NGB和HIF-1α阳性产物分布特征相似，且HIF-1α蛋白免疫阳性反应的整体强度高于NGB蛋白。各区域NGB和HIF-1α主要散在表达于特定层次的神经元胞质中，如延髓、脑桥神经元，小脑半球、蚓部皮质分子层、浦肯野细胞层及颗粒层神经元等。小脑半球髓质区的神经胶质细胞内也见有少量NGB和HIF-1α阳性表达，阴性对照均无表达。上述结果提示，牦牛后脑不同区域NGB和HIF-1α的表达在长期适应低氧环境过程中存在一定的选择性差异，蚓前叶对缺氧具有较高的耐受性，小脑半球皮质次之，蚓小叶的缺氧耐受性最弱，这种差异的产生可能与后脑相关区域承担的特定功能有关联，但其具体机制尚有待进一步探讨。各区域低氧适应性功能的发挥主要依赖其特定的神经元结构。

关键词: 脑红蛋白, 低氧诱导因子-1α, 牦牛, 后脑

Abstract: As the key factors related to neuroprotective function in the nervous system of vertebrates, neuroglobin (NGB) and hypoxia inducible factor-1α (HIF-1α) play important physiological functions in the process of hypoxic adaptation. This study aimed to explore the relationship between expression and distribution of NGB and HIF-1α in the process of adaptation to hypoxia of yak (Bos grunniens) hindbrain which plays an important regulatory function in animal movement, breathing, sensory and other physiological activities. RT-qPCR, Western blot and immunohistochemical methods were used to investigate the expression and localization characteristics of NGB and HIF-1α in different regions of yak hindbrain. The results showed that the expression trend characteristics between NGB and HIF-1α in yak hindbrain were basically consistent, the highest expression level was found in the archicerebellum which was significantly higher than those in the medulla oblongata, pons and other parts of cerebellum (P<0.05), the expression level in the cerebellar hemisphere cortex was less than that in the archicerebellum and the lowest expression level was found in the palaeocerebellum, there were also some differences in the expression levels of the two factors in other regions. The distribution characteristics of NGB and HIF-1α positive products were similar, and the overall intensity of the HIF-1α protein immunopositive response was higher than that of NGB protein. NGB and HIF-1α were mainly expressed in the cytoplasm of neurons in the medulla oblongata and pons. Scattered NGB and HIF-1α positive cells could be found in the molecular layer of the cerebellar cortex, archicerebellum, neocerebellum and palaeocerebellum. There were also a small number of positive expressions in the basket cells. The dendritic cell cytoplasm of Purkinje cells in the Purkinje cell layer showed a strong positive expression of NGB and HIF-1α. Strong positive expression of NGB and HIF-1α were also observed in the granular cell cytoplasm. The regions of white matter (medulla) showed rarely NGB and HIF-1α positive expression in the cytoplasm of the neurons. No expression was observed in the negative control. The above results suggested that there were selective differences between the expression of NGB and HIF-1α in different regions of yak hindbrain during the long-term adaptation to hypoxic environment. The archicerebellum has a higher tolerance to hypoxia, followed by the cerebellar hemisphere, and the weakest tolerance to hypoxia in the palaeocerebellum. The reason for the differences might be related to the specific functions undertaken by the relevant regions of the hindbrain in yak and the adaptive function of hypoxia in each region mainly depends on its specific neuronal structure.

Key words: neuroglobin, hypoxia inducible factor-1α, yak, hindbrain

中图分类号:

S852.16

杜晓华, 米晓钰, 王海芳, 温永强, 李乔, 刘霞. 脑红蛋白和缺氧诱导因子-1α在牦牛后脑的表达与定位研究[J]. 畜牧兽医学报, 2021, 52(3): 789-798.

DU Xiaohua, MI Xiaoyu, WANG Haifang, WEN Yongqiang, LI Qiao, LIU Xia. Expression and Localization of Neuroglobin and Hypoxia Inducible Factor-1α in Yak's Hindbrain[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 789-798.

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