Springer J E, Mu X, Bergmann L W, Trojanowski J Q
Department of Neurology, Hahnemann University School of Medicine, Philadelphia, Pennsylvania 19102-1192.
Exp Neurol. 1994 Jun;127(2):167-70. doi: 10.1006/exnr.1994.1091.
A recently cloned neurotrophic factor, termed glial cell line-derived neurotrophic factor (GDNF), has been reported to exhibit selective neurotrophic properties on ventral mesencephalon dopaminergic neurons, which degenerate in patients with Parkinson's disease. In the present study, we used reverse transcriptase followed by polymerase chain reaction (PCR) and in situ hybridization to study the expression of GDNF messenger RNA (mRNA) in the adult rat and human central nervous system (CNS). GDNF transcripts were identified using PCR in all regions of the rat CNS analyzed including striatum, hippocampus, cortex, cerebellum, and spinal cord. Interestingly, the rat hippocampal formation contained two transcripts, i.e., a larger form in addition to the amplified GDNF cDNA found in all other areas analyzed. GDNF PCR products also were observed in human striatum, hippocampus, cortex, and spinal cord, but not cerebellum, and both the striatum and hippocampal formation contained two GDNF transcripts. Finally, GDNF transcripts were detected in a rat Schwann cell line previously shown to secrete a factor that exerts a neurotrophic effect on dopaminergic neurons. In situ hybridization experiments using a cRNA probe hybridized to adult rat brain sections demonstrated no positive GDNF mRNA signal. However, intense GDNF mRNA hybridization signal was found to be associated with dorsal root ganglia in Postnatal Day 1 rats. These findings provide evidence that GDNF is detectable using PCR in a number of nervous system structures and, in some areas, GDNF is expressed in more than one form.
一种最近克隆的神经营养因子，称为胶质细胞系源性神经营养因子（GDNF），据报道对中脑腹侧多巴胺能神经元具有选择性神经营养特性，而这些神经元在帕金森病患者中会退化。在本研究中，我们使用逆转录酶随后进行聚合酶链反应（PCR）和原位杂交来研究GDNF信使核糖核酸（mRNA）在成年大鼠和人类中枢神经系统（CNS）中的表达。在分析的大鼠CNS的所有区域，包括纹状体、海马体、皮质、小脑和脊髓中，使用PCR鉴定出了GDNF转录本。有趣的是，大鼠海马结构包含两种转录本，即在所有其他分析区域中发现的扩增GDNF cDNA之外还有一种更大的形式。在人类纹状体、海马体、皮质和脊髓中也观察到了GDNF PCR产物，但在小脑中未观察到，并且纹状体和海马结构都包含两种GDNF转录本。最后，在先前显示能分泌对多巴胺能神经元发挥神经营养作用的因子的大鼠雪旺细胞系中检测到了GDNF转录本。使用与成年大鼠脑切片杂交的cRNA探针进行的原位杂交实验未显示阳性GDNF mRNA信号。然而，在出生后第1天的大鼠中，发现强烈的GDNF mRNA杂交信号与背根神经节相关。这些发现提供了证据，表明使用PCR可在许多神经系统结构中检测到GDNF，并且在某些区域，GDNF以不止一种形式表达。
