Kaldis P, Hemmer W, Zanolla E, Holtzman D, Wallimann T
Swiss Federal Institute of Technology, ETH-Hönggerberg, Zürich, Switzerland.
Dev Neurosci. 1996;18(5-6):542-54. doi: 10.1159/000111452.
Creatine kinase (CK) isoenzymes, with emphasis on the mitochondrial CK isoenzymes, were characterized and localized in chicken cerebellum. Chicken cerebellum extracts analyzed by two-dimensional gels, using antipeptide antibodies specific for sarcomeric muscle-type mitochondrial CK (Mib-CK) and revealed the presence of a Mib-CK variant in avian cerebellum. This CK isoform was localized by immunofluorescence staining exclusively in the Purkinje neurons. The co-expression of this Mib-CK together with cytosolic muscle-type MM-CK, as observed in the same Purkinje neurons, may reflect the specific energy requirements associated with highly fluctuating Ca2+ levels (Ca2+ spiking) in these specialized neurons. Ubiquitous brain-type mitochondrial Mia-CK was found together with cytosolic BB-CK mainly in the glomeruli structures of the cerebellar granular layer. BB-CK, but much less so Mia-CK however, was also very prominent in Bergmann glial cells of the two mitochondrial Mi-CK isoenzymes in the chicken cerebellum is demonstrated. Other hot spots of CK localization were the granule and pyramidal cells of the hippocampus in rat. There, a developmental stage-dependent immunofluorescence staining, especially with antibodies against Mia-CK was noted. Epithelial cells of the choroid plexus were also highly enriched in CK. The possible implications of a CK/PCr circuit at these various cellular locations of the brain are discussed with respect to normal brain physiology and pathology.
对肌酸激酶(CK)同工酶进行了表征,并将其定位于鸡小脑,重点研究了线粒体CK同工酶。通过二维凝胶分析鸡小脑提取物,使用针对肌节肌型线粒体CK(Mib-CK)的抗肽抗体,发现禽类小脑中存在Mib-CK变体。通过免疫荧光染色将这种CK同工型仅定位于浦肯野神经元。在同一浦肯野神经元中观察到这种Mib-CK与胞质肌型MM-CK的共表达,可能反映了这些特殊神经元中与高度波动的Ca2+水平(Ca2+尖峰)相关的特定能量需求。普遍存在的脑型线粒体Mia-CK与胞质BB-CK主要在小脑颗粒层的肾小球结构中被发现。BB-CK在鸡小脑的两种线粒体Mi-CK同工酶的伯格曼胶质细胞中也非常突出,而Mia-CK则较少。CK定位的其他热点是大鼠海马体的颗粒细胞和锥体细胞。在那里,观察到了发育阶段依赖性免疫荧光染色,尤其是针对Mia-CK的抗体。脉络丛上皮细胞中CK也高度富集。就正常脑生理学和病理学而言,讨论了CK/磷酸肌酸循环在脑的这些不同细胞位置的可能影响。