The chemokine growth-related gene product β protects rat cerebellar granule cells from apoptotic cell death through α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptors
The chemokine growth-related gene product β protects rat cerebellar granule cells from apoptotic cell death through α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptors
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DOI：
10.1073/pnas.090105997
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发表时间：
2000-05
影响因子：
11.1
通讯作者：
C. Limatola;M. Ciotti;D. Mercanti;F. Vacca;D. Ragozzino;A. Giovannelli;A. Santoni;F. Eusebi;R. Miledi
中科院分区：
文献类型：
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作者：
C. Limatola;M. Ciotti;D. Mercanti;F. Vacca;D. Ragozzino;A. Giovannelli;A. Santoni;F. Eusebi;R. Miledi
培养的小脑颗粒神经元被广泛用作研究神经细胞死亡机制的细胞模型，因为当它们从含25 mM K+的培养液切换到含5 mM K+的培养液时，它们会发生程序性细胞死亡。我们发现生长相关基因产物谷氨酸(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate，β)可部分阻止K+耗竭所致的细胞死亡，Groβ对颗粒细胞的神经保护作用是通过AMPA型离子亲和型谷氨酸受体介导的。β/海人藻酸受体的特异性拮抗剂6-氰基-7-硝基喹恶啉-2，3-二酮可抑制GRO-GRO诱导的细胞存活，但不受N-甲基-D-天冬氨酸受体抑制剂2-氨基-5-磷酸戊酸的影响，其作用与AMPA(10μM)诱导的颗粒细胞存活相当。此外，当颗粒细胞被针对β受体亚基的反义寡核苷酸处理时，Gro AMPA诱导的神经保护作用被取消，这显著降低了受体的表达，亚基特异性抗体的Western印迹分析和颗粒细胞对AMPA的电生理敏感性证实了这一点。我们的数据表明，GROβ对小脑颗粒细胞具有神经营养作用，并且这种活性依赖于AMPA受体。
Cultured cerebellar granule neurons are widely used as a cellular model to study mechanisms of neuronal cell death because they undergo programmed cell death when switched from a culture medium containing 25 mM to one containing 5 mM K+. We have found that the growth-related gene product β (GROβ) partially prevents the K+-depletion-induced cell death, and that the neuroprotective action of GROβ on granule cells is mediated through the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) type of ionotropic glutamate receptors. GROβ-induced survival was suppressed by 6-cyano-7-nitroquinoxaline-2,3-dione, which is a specific antagonist of AMPA/kainate receptors; it was not affected by the inhibitor of N-methyl-d-aspartate receptors, 2-amino-5-phosphonopentanoic acid, and was comparable to the survival of granule cells induced by AMPA (10 μM) treatment. Moreover, GROβ-induced neuroprotection was abolished when granule cells were treated with antisense oligonucleotides specific for the AMPA receptor subunits, which significantly reduced receptor expression, as verified by Western blot analysis with subunit-specific antibodies and by granule cell electrophysiological sensitivity to AMPA. Our data demonstrate that GROβ is neurotrophic for cerebellar granule cells, and that this activity depends on AMPA receptors.