Neural stem cells express melatonin receptors and neurotrophic factors: colocalization of the MT1 receptor with neuronal and glial markers.
Neural stem cells express melatonin receptors and neurotrophic factors: colocalization of the MT1 receptor with neuronal and glial markers.
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DOI：
10.1186/1471-2202-5-41
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发表时间：
2004-10-28
期刊：
影响因子：
2.4
通讯作者：
Kirkham DL
中科院分区：
文献类型：
--
作者：
Niles LP;Armstrong KJ;Rincón Castro LM;Dao CV;Sharma R;McMillan CR;Doering LC;Kirkham DL
为了优化神经干细胞（NSC）移植治疗神经退行性疾病的潜在益处，有必要了解其生物学特性。虽然神经营养因子转导策略是有希望的，但替代方法如通过NSC调节内源性神经营养因子表达也可能是有益的。因此，利用C17.2神经干细胞系，我们研究了在不同的体外条件下所选择的神经营养因子的表达。鉴于最近的证据表明松果体激素褪黑激素在脊椎动物发育中的作用，确定其G蛋白偶联的MT 1和MT 2受体是否在NSC中表达也是令人感兴趣的。RT-PCR分析显示，在培养中维持两天的未分化细胞中，胶质细胞系源性神经营养因子（GDNF）、脑源性神经营养因子（BDNF）和神经生长因子（NGF）的稳健表达。一周后，分化中的细胞继续表现出BDNF和NGF的高表达，但GDNF表达较低或不表达，这取决于所用的培养条件。在培养两天的神经干细胞中检测到褪黑素MT 1受体mRNA，但未见MT 2受体。在培养两天的神经干细胞中通过蛋白质印迹法检测到约30 kDa的未成熟MT 1受体，而在维持较长时间的细胞中存在约40 - 45 kDa的成熟受体。免疫细胞化学研究表明，MT 1受体在神经（β-微管蛋白III阳性）和胶质（GFAP阳性）祖细胞中表达。褪黑激素对神经营养因子表达的影响的检查显示，低生理浓度的这种激素引起治疗24小时后神经干细胞中GDNF mRNA表达的显著诱导。C17.2细胞的表型特征表明它们是包括神经和神经胶质祖细胞的NSC的异质群体，如在本研究中使用的细胞培养条件下观察到的。这些神经干细胞具有表达神经营养因子的内在能力，在培养数天后明显抑制GDNF表达。褪黑激素受体在神经干/祖细胞的检测表明，这种多效性激素在哺乳动物神经发育的参与。此外，褪黑激素诱导GDNF在C17.2细胞中表达的能力支持MT 1受体在这些NSC中表达的功能作用。鉴于GDNF在促进多巴胺能神经元存活方面的潜力，这些新的发现对褪黑激素在神经保护策略中的利用具有影响，特别是在帕金森病中。
In order to optimize the potential benefits of neural stem cell (NSC) transplantation for the treatment of neurodegenerative disorders, it is necessary to understand their biological characteristics. Although neurotrophin transduction strategies are promising, alternative approaches such as the modulation of intrinsic neurotrophin expression by NSCs, could also be beneficial. Therefore, utilizing the C17.2 neural stem cell line, we have examined the expression of selected neurotrophic factors under different in vitro conditions. In view of recent evidence suggesting a role for the pineal hormone melatonin in vertebrate development, it was also of interest to determine whether its G protein-coupled MT1 and MT2 receptors are expressed in NSCs. RT-PCR analysis revealed robust expression of glial cell-line derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in undifferentiated cells maintained for two days in culture. After one week, differentiating cells continued to exhibit high expression of BDNF and NGF, but GDNF expression was lower or absent, depending on the culture conditions utilized. Melatonin MT1 receptor mRNA was detected in NSCs maintained for two days in culture, but the MT2 receptor was not seen. An immature MT1 receptor of about 30 kDa was detected by western blotting in NSCs cultured for two days, whereas a mature receptor of about 40 – 45 kDa was present in cells maintained for longer periods. Immunocytochemical studies demonstrated that the MT1 receptor is expressed in both neural (β-tubulin III positive) and glial (GFAP positive) progenitor cells. An examination of the effects of melatonin on neurotrophin expression revealed that low physiological concentrations of this hormone caused a significant induction of GDNF mRNA expression in NSCs following treatment for 24 hours. The phenotypic characteristics of C17.2 cells suggest that they are a heterogeneous population of NSCs including both neural and glial progenitors, as observed under the cell culture conditions used in this study. These NSCs have an intrinsic ability to express neurotrophic factors, with an apparent suppression of GDNF expression after several days in culture. The detection of melatonin receptors in neural stem/progenitor cells suggests involvement of this pleiotropic hormone in mammalian neurodevelopment. Moreover, the ability of melatonin to induce GDNF expression in C17.2 cells supports a functional role for the MT1 receptor expressed in these NSCs. In view of the potency of GDNF in promoting the survival of dopaminergic neurons, these novel findings have implications for the utilization of melatonin in neuroprotective strategies, especially in Parkinson's disease.
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影响因子：
1.7
作者：
Al-Ghoul, WM;Herman, MD;Dubocovich, ML
通讯作者：
Dubocovich, ML
影响因子：
5.3
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Åkerud, P;Canals, JM;Arenas, E
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Arenas, E
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2.2
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Abecia, JA;Forcada, F;Zúñiga, O
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Zúñiga, O
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56.9
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LIN, LFH;DOHERTY, DH;COLLINS, F
通讯作者：
COLLINS, F
影响因子：
3.8
作者：
Overbergh, L;Valckx, D;Mathieu, C
通讯作者：
Mathieu, C