Differential regulation of glial cell line‐derived neurotrophic factor (GDNF) mRNA expression during hypoxia and reoxygenation in astrocytes isolated from stroke‐prone spontaneously hypertensive rats
Differential regulation of glial cell line‐derived neurotrophic factor (GDNF) mRNA expression during hypoxia and reoxygenation in astrocytes isolated from stroke‐prone spontaneously hypertensive rats
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DOI：
10.1002/glia.10003
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发表时间：
2002-01
期刊：
影响因子：
6.2
通讯作者：
K. Yamagata;M. Tagami;K. Ikeda;S. Tsumagari;Y. Yamori;Y. Nara
中科院分区：
文献类型：
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作者：
K. Yamagata;M. Tagami;K. Ikeda;S. Tsumagari;Y. Yamori;Y. Nara
胶质细胞源性神经营养因子（GDNF）在缺血期间成熟神经元的存活和恢复中发挥着多种重要作用。我们研究了以下可能性：在缺氧和复氧 (H/R) 期间以及暴露于谷氨酸和过氧化氢 (H2O2) 期间，与 Wistar 京都大鼠 (WKY) 相比，易中风自发性高血压大鼠 (SHRSP) 培养的星形胶质细胞中 GDNF mRNA 的表达和 GDNF 蛋白的释放受到差异性调节。基于荧光 TaqMan 方法，通过逆转录聚合酶链反应（RT-PCR）对 mRNA 表达进行定量。一种能够实时测量荧光的新仪器被用来量化星形胶质细胞的基因扩增。通过酶联免疫吸附测定（ELISA）研究GDNF蛋白。正常氧条件下，SHRSP 中的 GDNF mRNA 表达和 GDNF 蛋白释放量高于 WKY 星形胶质细胞。然而，在 H/R 期间，与 WKY 相比，SHRSP 的 mRNA 表达和蛋白质释放趋于减少。谷氨酸和 H2O2 以剂量依赖性方式诱导 WKY 和 SHRSP 中 GDNF mRNA 的表达和 GDNF 蛋白的释放。 SHRSP 中的 GDNF mRNA 和蛋白水平显着低于 WKY。这些发现表明，与 WKY 中观察到的相比，SHRSP 星形胶质细胞中对 H/R、谷氨酸和 H2O2 的 GDNF 产生较低。我们得出的结论是，星形胶质细胞中 GDNF 产生的减弱与 H/R 期间 SHRSP 的神经元脆弱性有关，因为谷氨酸和 H2O2 刺激的 GDNF 产生与针对 H/R 介导的神经毒性的保护作用密切相关。 GLIA 37:1–7, 2002。© 2002 Wiley‐Liss, Inc.
Glial cell line‐derived neurotrophic factor (GDNF) plays several important roles in the survival and recovery of mature neurons during ischemia. We examined the possibility that the expression of GDNF mRNA and the release of GDNF protein are regulated differentially in cultured astrocytes from the stroke‐prone spontaneously hypertensive rat (SHRSP) compared with those from Wistar Kyoto rats (WKY) during hypoxia and reoxygenation (H/R) and after exposure to glutamate and hydrogen peroxide (H2O2). The mRNA expression was quantitated by reverse transcription‐polymerase chain reaction (RT‐PCR) based on the fluorescent TaqMan methodology. A new instrument capable of measuring fluorescence in real‐time was used to quantify gene amplification in astrocytes. GDNF protein was investigated by enzyme‐linked immunosorbent assay (ELISA). GDNF mRNA expression and GDNF protein release at normoxia were greater in SHRSP than in WKY astrocytes. During H/R, however, the mRNA expression and protein release tended to be reduced in SHRSP compared with WKY. Glutamate and H2O2 induced the expression of GDNF mRNA and the release of GDNF protein in both WKY and SHRSP in a dose‐dependent manner. Levels of GDNF mRNA and protein in SHRSP were significantly lower than in WKY. These findings indicate that GDNF production in SHRSP astrocytes was low in response to H/R, glutamate, and H2O2, compared with that observed in WKY. We conclude that the attenuated production of GDNF in astrocytes is involved in neuronal vulnerability in SHRSP during H/R, as GDNF production, which is stimulated by glutamate and H2O2, is closely related to the protective effect against H/R‐mediated neurotoxicity. GLIA 37:1–7, 2002. © 2002 Wiley‐Liss, Inc.