EXPRESSION OF TYROSINE-HYDROXYLASE IN CEREBELLAR PURKINJE NEURONS OF THE MUTANT TOTTERING AND LEANER MOUSE
EXPRESSION OF TYROSINE-HYDROXYLASE IN CEREBELLAR PURKINJE NEURONS OF THE MUTANT TOTTERING AND LEANER MOUSE
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DOI：
10.1016/0169-328x(92)90113-p
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发表时间：
1992-10-01
期刊：
影响因子：
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通讯作者：
CRAWLEY, JN
中科院分区：
文献类型：
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作者：
AUSTIN, MC;SCHULTZBERG, M;CRAWLEY, JN
采用原位杂交组织化学、北方印迹分析和免疫组织化学方法，检测了C57 BL/6 J背景下饲养的蹒跚（tg/tg）、消瘦（tg（la）/tg（la））、复合杂合子（tg/tg（la））和野生型对照（+/+）小鼠蓝斑和小脑中酪氨酸羟化酶（TH）mRNA的含量和免疫反应性。小脑浦肯野神经元，长期以来被认为是GABA能的，在tg/tg，tg/tg（la）和tg（la）/tg（la）小鼠的尾蚓部和小脑侧半球显示高水平的TH mRNA。对单个浦肯野细胞的颗粒密度分析显示，与+/+野生型对照小鼠相比，tg/tg、tg/tg（la）和tg（la）/tg（la）小鼠中TH mRNA的浓度显著更高。成年（大于或等于2个月）和年轻的，癫痫发作前（小于或等于3周）突变小鼠的比较表明，浦肯野细胞密集标记TH mRNA在两个年龄，表明TH基因表达在浦肯野细胞是独立的癫痫发作。北方印迹分析证实了原位杂交研究的结果，显示了与TH mRNA相同的单一条带。免疫组化证实TH蛋白的存在下，在浦肯野细胞的尾蚓部和小脑的侧半球在控制和突变小鼠。在成年tg/tg、tg/tg（la）和+/+对照小鼠之间，蓝斑中TH和共存的神经肽甘丙肽的mRNA的定量没有检测到显著差异。目前的研究结果表明，经典的GABA能浦肯野细胞在小脑表达低水平的TH，和突变摇摇欲坠和瘦品系的小鼠表达极高水平的TH的mRNA和蛋白质。
In situ hybridization histochemistry, Northern blot analysis and immunohistochemistry were used to examine tyrosine hydroxylase (TH) mRNA concentrations and immunoreactivity in the locus coeruleus and cerebellum of the tottering (tg/tg), leaner (tg(la)/tg(la)), compound heterozygous (tg/tg(la)) and wild type control (+/+) mice, bred on a C57BL/6J background. Cerebellar Purkinje neurons, long considered to be GABAergic, showed high levels of TH mRNA in the caudal vermis and the lateral hemispheres of the cerebellum of tg/tg, tg/tg(la), and tg(la)/tg(la) mice. Analysis of grain density over individual Purkinje cells showed significantly greater concentrations of TH mRNA in tg/tg, tg/tg(la), and tg(la)/tg(la) mice as compared to +/+ wild type control mice. Comparison of adult (greater-than-or-equal-to 2 months) and young, pre-seizure (less-than-or-equal-to 3 weeks) mutant mice showed Purkinje cells densely labelled for TH mRNA at both ages, suggesting that TH gene expression in Purkinje cells is independent of the onset of seizures. Northern blot analysis confirmed the findings from the in situ hybridization studies, demonstrating a single band identical to TH mRNA. Immunohistochemistry confirmed the presence of TH protein in Purkinje cells of the caudal vermis and the lateral hemispheres of the cerebellum in both control and mutant mice. Quantitation of mRNA for TH and the coexisting neuropeptide, galanin, in the locus coeruleus detected no significant differences between adult tg/tg, tg/tg(la) and +/+ control mice. The present findings demonstrate that the classically GABAergic Purkinje cells in the cerebellum express low levels of TH, and that the mutant tottering and leaner strains of mice express extremely high levels of mRNA and protein for TH.