Liu Zhiping, Martin Lee J
Department of Pathology, Division of Neuropathology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
J Comp Neurol. 2003 May 12;459(4):368-91. doi: 10.1002/cne.10664.
The olfactory bulb (OB) core is an extension of the rostral migratory stream and thus is a potential source of neural progenitor and neural stem cells. We characterized in vivo and in vitro neuronal progenitor and neural stem cells in the adult OB core. In mouse and rat, bromodeoxyuridine (BrdU) labeling showed that the OB core accumulates newly replicated cells. Nestin, a neuroepithelial stem cell marker, was enriched in the OB core. BrdU-positive cells were immunolabeled for nestin and TUC4, a marker for early postmitotic neurons. The distributions of cells labeled for BrdU, TUC4, and nestin were similarly concentrated in the OB core. Nestin- and TUC4-positive cells were also found in the OB of young and aged humans. Isolated and cultured OB core cells from adult rat and mouse had the capacity to generate numerous neurospheres. Adult OB core neurospheres were cryopreserved and subsequently cultured. Single cell clonal analysis of neurospheres revealed the capacity for self-renewal and multipotency. Cultured adult OB core cells differentiated into neurons, astrocytes, and oligodendrocytes. Some neurons expressed choline acetlytransferase, substance P, and glutamic acid decarboxylase. Basic fibroblast growth factor potentiated the self-renewal of cells and beta-nerve growth factor stimulated differentiation. OB-derived neural stem cells in coculture with skeletal muscle cells were induced to become neurons expressing choline acetyltransferase and substance P and formed neuromuscular synaptic junctions on myocytes displaying acetylcholinesterase-positive motor end plates. Cocultured OB-derived neural stem cells with myoblast cells also generated nonneural cell progeny. We conclude that the adult mammalian OB core is a reservoir of neural progenitor cells and pluripotent neural stem cells.
嗅球(OB)核心是吻侧迁移流的延伸部分,因此是神经祖细胞和神经干细胞的潜在来源。我们对成年OB核心中的神经元祖细胞和神经干细胞进行了体内和体外特征分析。在小鼠和大鼠中,溴脱氧尿苷(BrdU)标记显示OB核心积累了新复制的细胞。巢蛋白,一种神经上皮干细胞标志物,在OB核心中富集。BrdU阳性细胞用巢蛋白和TUC4进行免疫标记,TUC4是有丝分裂后早期神经元的标志物。标记有BrdU、TUC4和巢蛋白的细胞分布同样集中在OB核心。在年轻和老年人类的OB中也发现了巢蛋白和TUC4阳性细胞。从成年大鼠和小鼠中分离并培养的OB核心细胞有能力产生大量神经球。成年OB核心神经球被冷冻保存并随后进行培养。对神经球的单细胞克隆分析揭示了其自我更新和多能性的能力。培养的成年OB核心细胞分化为神经元、星形胶质细胞和少突胶质细胞。一些神经元表达胆碱乙酰转移酶、P物质和谷氨酸脱羧酶。碱性成纤维细胞生长因子增强细胞的自我更新,β-神经生长因子刺激分化。与骨骼肌细胞共培养的OB来源的神经干细胞被诱导成为表达胆碱乙酰转移酶和P物质的神经元,并在显示乙酰胆碱酯酶阳性运动终板的肌细胞上形成神经肌肉突触连接。与成肌细胞共培养的OB来源的神经干细胞也产生非神经细胞后代。我们得出结论,成年哺乳动物的OB核心是神经祖细胞和多能神经干细胞的储存库。