Breese C R, Leonard S S
Department of Pharmacology, University of Colorado Health Sciences Center, Denver 80262.
J Mol Neurosci. 1993 Winter;4(4):263-75. doi: 10.1007/BF02821558.
Antibodies to functional glutamate receptor subunits were utilized as probes to characterize glutamatergic receptors in human postmortem brain tissue. Crude membranes from rat, monkey, and various dissected human postmortem brain regions were fractionated by SDS-PAGE and electrotransferred to nitrocellulose. Using antisera raised against rat antigens for AMPA/kainate (GluR1-3) and kainate (GluR5) glutamate receptor subunits, we have been able to detect specific bands on Western blots in rat, monkey, and human postmortem brain tissue. These antisera recognized bands at approx 105 kDa for the GluR1-3 and 115 kDa for GluR5 in humans, monkeys, and rats. All of these glutamate receptor subtypes appear to be glycosylated. We observed varying levels of expression in the human brain areas examined, with the highest degree of expression in the hippocampus and temporal cortex for AMPA/kainate receptor subunits, and in the cortex and cerebellum for the kainate receptor subunits. In addition, considerable heterogeneity in expression was observed between protein, NCAM. Our studies indicate that glutamatergic receptor protein changes related to various human diseases states may be examined in human postmortem tissue by Western blotting techniques utilizing these antibodies raised to the rat protein.
利用针对功能性谷氨酸受体亚基的抗体作为探针,对人死后脑组织中的谷氨酸能受体进行表征。来自大鼠、猴子和各种解剖的人死后脑区的粗制膜通过SDS-PAGE进行分级分离,并电转移至硝酸纤维素膜上。使用针对大鼠抗原产生的抗血清,用于检测AMPA/海人藻酸(GluR1-3)和海人藻酸(GluR5)谷氨酸受体亚基,我们已经能够在大鼠、猴子和人死后脑组织的蛋白质印迹上检测到特异性条带。这些抗血清在人、猴和大鼠中识别出GluR1-3约105 kDa和GluR5约115 kDa的条带。所有这些谷氨酸受体亚型似乎都被糖基化。我们在所检查的人脑区域中观察到不同的表达水平,AMPA/海人藻酸受体亚基在海马体和颞叶皮质中表达程度最高,而海人藻酸受体亚基在皮质和小脑中表达程度最高。此外,在蛋白质、NCAM之间观察到表达的显著异质性。我们的研究表明,利用针对大鼠蛋白质产生的这些抗体,通过蛋白质印迹技术可以在人死后组织中检测与各种人类疾病状态相关的谷氨酸能受体蛋白变化。