Labatut R, Buda M, Bérod A
INSERM U 171, CNRS UA 1195, Centre Hospitalier Lyon-Sud, Pierre Bénite, France.
J Neurochem. 1988 May;50(5):1375-80. doi: 10.1111/j.1471-4159.1988.tb03019.x.
An immunoblot procedure was developed to quantify the amount of tyrosine hydroxylase protein in homogenate of small brain regions. With the use of this method we have studied the variations in tyrosine hydroxylase activity and protein levels in some catecholaminergic neurons at different times following a single reserpine injection (10 mg/kg s.c.) and reevaluated the anatomical specificity of tyrosine hydroxylase induction by this drug. Reserpine administration provoked a long-lasting increase in both tyrosine hydroxylase activity and protein levels within locus ceruleus neurons. This effect culminated at day 4 after injection. At this time, the enzyme activity and protein levels in treated animals were respectively 2.7 and 2.6 times that measured in vehicle-treated animals. Both parameters varied in parallel so that tyrosine hydroxylase specific activity did not change over time. In contrast, reserpine did not cause any changes in tyrosine hydroxylase activity in the dopaminergic neurons of the substantia nigra, but provoked a moderate increase in tyrosine hydroxylase protein level. This latter effect was maximal (1.5 times) 4 days after treatment. In the adjacent dopaminergic area, i.e., the ventral tegmental area, a small decrease in the enzyme activity was recorded at day 2 without any significant change in the level of the protein. In conclusion, first, our data show the capacity of our method to assay tyrosine hydroxylase protein amounts in small brain catecholaminergic nuclei. Second, our results confirm and extend previous studies on the effect of reserpine on the regulation of tyrosine hydroxylase level within brain noradrenergic neurons.(ABSTRACT TRUNCATED AT 250 WORDS)
我们开发了一种免疫印迹法来定量测定小脑小区域匀浆中酪氨酸羟化酶蛋白的含量。利用该方法,我们研究了单次注射利血平(10 mg/kg,皮下注射)后不同时间一些儿茶酚胺能神经元中酪氨酸羟化酶活性和蛋白水平的变化,并重新评估了该药物诱导酪氨酸羟化酶的解剖学特异性。给予利血平后,蓝斑核神经元内酪氨酸羟化酶活性和蛋白水平均出现持久升高。这种效应在注射后第4天达到顶峰。此时,处理组动物的酶活性和蛋白水平分别是溶剂处理组动物的2.7倍和2.6倍。两个参数平行变化,因此酪氨酸羟化酶比活性没有随时间变化。相比之下,利血平对黑质多巴胺能神经元的酪氨酸羟化酶活性没有任何影响,但引起酪氨酸羟化酶蛋白水平适度升高。后一种效应在处理后4天最大(1.5倍)。在相邻的多巴胺能区域,即腹侧被盖区,第2天记录到酶活性略有下降,而蛋白水平没有显著变化。总之,首先,我们的数据表明我们的方法能够测定小脑儿茶酚胺能核团中酪氨酸羟化酶蛋白的含量。其次,我们的结果证实并扩展了先前关于利血平对脑内去甲肾上腺素能神经元中酪氨酸羟化酶水平调节作用的研究。(摘要截短至250字)