Calretinin and calbindin D-28k delay the onset of cell death after excitotoxic stimulation in transfected P19 cells
Calretinin and calbindin D-28k delay the onset of cell death after excitotoxic stimulation in transfected P19 cells
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DOI：
10.1016/s0006-8993(01)02671-3
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发表时间：
2001-08-03
期刊：
影响因子：
2.9
通讯作者：
Celio, MR
中科院分区：
文献类型：
--
作者：
D'Orlando, C;Fellay, B;Celio, MR
在某些神经系统疾病中，神经元的损伤反映了兴奋性氨基酸对其受体的过度刺激。这种反应可能会扰乱Ca 2+稳态，并导致该离子的细胞内浓度显著和持续增加。根据相关研究的数据，钙结合蛋白被认为在这些病理中起保护作用。我们直接测试了三种钙结合蛋白calretinin（CR）、calbindin D-28 k（CB）和parvalbumin（PV）缓冲[Ca 2 +]和保护细胞免受兴奋性毒性死亡的能力。我们使用P19小鼠胚胎癌细胞，它可以特异性地诱导（维甲酸）转化为神经样细胞。分化的细胞表达功能性谷氨酸受体，易受兴奋性毒性休克的影响。用诱导分化的CR、CB或PV的cDNA稳定转染未分化的P19细胞，然后暴露于谷氨酸受体激动剂NMDA。表达CR，CB或PV的克隆的存活率进行了比较与未转染的P19细胞使用乳酸脱氢酶测定。CR-和CB-表达细胞在暴露于NMDA的第一个2小时内免于死亡。然而，这种保护是短暂的，并不足以拯救P19-细胞长时间刺激后。提出的三个PV转染的克隆中的两个容易受到NMDA诱导的兴奋性毒性;第三个，表达最低水平的PV，被保护到与CR和CB转染的克隆相似的程度。我们的研究结果表明，在P19细胞模型中，CR和CB可以帮助延迟兴奋性毒性刺激后细胞死亡的发生。(C)2001 Elsevier Science B. V.保留所有权利。
In some neurological diseases, injury to neurones reflects an over-stimulation of their receptors for excitatory amino acids. This response may disturb the Ca2+-homeostasis and lead to a pronounced and sustained increase in the intracellular concentration of this ion. On the basis of data derived from correlative studies, calcium-binding proteins have been postulated to play a protective role in these pathologies. We tested, directly, the capacity of the three calcium-binding proteins calretinin (CR), calbindin D-28k (CB) and parvalbumin (PV) to buffer [Ca2+], and to protect cells against excitotoxic death. We used P19 murine embryonic carcinoma cells, which can be specifically induced (by retinoic acid) to transform into nerve-like ones. The differentiated cells express functional glutamate-receptors and are susceptible to excitotoxic shock. Undifferentiated P19-cells were stably transfected with the cDNA for CR, CB or PV induced to differentiate, and then exposed to NMDA, a glutamate-receptor agonist. The survival rates of clones expressing CR, CB or PV were compared with those of untransfected P19-cells using the lactate-dehydrogenase assay. CR- and CB-expressing cells were protected from death during the first 2 h of exposure to NMDA. This protection was, however, transient, and did not suffice to rescue P19-cells after prolonged stimulation. Two of the three PV-transfected clones raised were vulnerable to NMDA-induced excitotoxicity; the third, which expressed the lowest level of PV, was protected to a similar degree as that found for the CR- and CB-transfected clones. Our results indicate that in the P19-cell model, CR and CB can help to delay the onset of cell death after excitotoxic stimulation. (C) 2001 Elsevier Science B.V. All rights reserved.