Department of Anatomy and Neurobiology, Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan 2nd Road, Guangzhou, 510080, PR China.
Neurochem Int. 2010 Apr;56(5):694-702. doi: 10.1016/j.neuint.2010.02.005. Epub 2010 Feb 11.
This study explores recently identified neurons that express the protein nestin in the medial septum-diagonal band of Broca (MS-DBB) of adult rats and humans. These nestin positive neurons from MS-DBB are known to project to the hippocampus and frontal cortex of the brain. However, their chemical identification has not been fully elucidated. In this study, we further investigated the chemical identity of the nestin-immunoreactive (ir) neurons in rats using double immunofluorescence labeling and single cell reverse transcription polymerase chain reaction (RT-PCR) techniques. The results of double labeling showed that all nestin-ir neurons exhibited choline acetyltransferase (ChAT) immunoreactivity in the MS-DBB of the basal forebrain. Conversely, only about 43% of the ChAT-ir neurons showed nestin immunoreactivity. In addition, a vast majority of the nestin-ir neurons (95%) were nerve growth factor receptor (NGFR) positive. The nestin-ir neurons were highly distributed in the rostral and intermediated regions of the MS-DBB. Single cell RT-PCR results showed that 90% of the nestin mRNA expressed neurons displayed ChAT mRNA expression as well, but neither the mRNA of the proteins glutamic acid decarboxylases 67 (GAD67) nor vesicular glutamate transporters (VGLUTs). These results provide the first evidence that nestin-ir neurons in the basal forebrain are a subpopulation of the classic cholinergic neurons. With high NGFR proteins activated in the nestin-ir neurons, we propose that the presence of nestin in the cholinergic neurons might be related to the survival and plasticity of the cholinergic neurons.
本研究探索了在成年大鼠和人类的脑内侧隔核-斜角带核（MS-DBB）中表达蛋白巢蛋白的新鉴定神经元。已知来自 MS-DBB 的巢蛋白阳性神经元投射到大脑的海马体和额叶皮质。然而，它们的化学鉴定尚未完全阐明。在这项研究中，我们使用双重免疫荧光标记和单细胞逆转录聚合酶链反应（RT-PCR）技术进一步研究了大鼠 MS-DBB 中巢蛋白免疫反应性（ir）神经元的化学特性。双重标记的结果表明，所有巢蛋白-ir 神经元在基底前脑的 MS-DBB 中均表现出胆碱乙酰转移酶（ChAT）免疫反应性。相反，只有约 43%的 ChAT-ir 神经元显示巢蛋白免疫反应性。此外，绝大多数巢蛋白-ir 神经元（95%）为神经生长因子受体（NGFR）阳性。巢蛋白-ir 神经元在 MS-DBB 的头侧和中间区域高度分布。单细胞 RT-PCR 结果表明，90%表达巢蛋白 mRNA 的神经元也表达 ChAT mRNA，但谷氨酸脱羧酶 67（GAD67）和囊泡谷氨酸转运体（VGLUTs）的蛋白质的 mRNA 均不表达。这些结果首次提供了证据表明，基底前脑的巢蛋白-ir 神经元是经典胆碱能神经元的一个亚群。由于巢蛋白-ir 神经元中存在高表达的 NGFR 蛋白，我们提出胆碱能神经元中巢蛋白的存在可能与胆碱能神经元的存活和可塑性有关。
