Transient Recovery in a Rat Model of Familial Amyotrophic Lateral Sclerosis After Transplantation of Motor Neurons Derived From Mouse Embryonic Stem Cells
Transient Recovery in a Rat Model of Familial Amyotrophic Lateral Sclerosis After Transplantation of Motor Neurons Derived From Mouse Embryonic Stem Cells
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DOI：
10.3727/096368909x12483162197123
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发表时间：
2009-01-01
影响因子：
3.3
通讯作者：
Velasco, Ivan
中科院分区：
文献类型：
--
作者：
Lopez-Gonzalez, Rodrigo;Kunckles, Philip;Velasco, Ivan
胚胎干细胞可以被诱导分化为运动神经元。类似于在家族性肌萎缩侧索硬化症（ALS）中观察到的MN变性和瘫痪的动物模型先前已被报道。在这项工作中，我们的目的是调查是否移植MN可以防止转基因大鼠表达的突变形式的人超氧化物歧化酶I（hSOD 1（G93 A））与遗传性ALS相关的运动退化。将小鼠ES细胞分化为在MN特异性基因09的启动子下表达绿色荧光蛋白（GFP）以及指示MN身份的分子标记的神经元。将细胞移植到10周龄的成年野生型（WT）或hSOD 1（G93 A）大鼠的腰脊髓中，此时转基因动物是症状前的。具有MN表型的移植细胞可以在hSOD 1（G93 A）动物中存活至少1周。为了定量评估WT和转基因大鼠的运动表现，我们在动物14周龄时开始每周进行转棒测试。假手术和移植WT动物在旋转杆上维持自身的能力没有下降。相比之下，假hSOD 1（G93 A）大鼠从第16周开始运动表现下降，到第19周达到瘫痪。在移植的转基因动物中，当与假hSOD 1（G93 A）相比时，在第16周和第17周旋转棒能力有显著改善。然而，在接下来的几周内，移植hSOD 1（G93 A）大鼠表现出运动恶化，并最终在第19周表现出瘫痪。在终末期，我们发现只有少数内源性MN假手术和移植hSOD 1（G93 A）大鼠通过甲酚紫染色;没有胆碱乙酰转移酶阳性或GFP阳性MN存在于移植转基因受试者。相比之下，在相同年龄分析的WT大鼠在其脊髓中具有移植的GFP阳性MN。这些结果强烈表明，转基因hSOD 1（G93 A）的环境是有害的，从长远来看，移植MN。
Embryonic stem (ES) cells can be induced to differentiate into motor neurons (MN). Animal models resembling MN degeneration and paralysis observed in familial amyotrophic lateral sclerosis (ALS) have been previously reported. In this work, we aimed to investigate whether transplanted MN could prevent motor deterioration in transgenic rats expressing a mutant form of human superoxide dismutase I (hSOD1(G93A)) associated with inherited ALS. Mouse ES cells were differentiated to neurons that express green fluorescent protein (GFP) under the promoter of the MN-specific gene 09, as well as molecular markers indicative of MN identity. Cells were grafted into the lumbar spinal cord of adult wild-type (WT) or hSOD1(G93A) rats at 10 weeks of age, when transgenic animals are presymptomatic. Grafted cells with MN phenotype can survive for at least I week in hSOD1(G93A) animals. To quantitatively evaluate motor performance of WT and transgenic rats, we carried out weekly rotarod tests starting when the animals were 14 weeks old. Sham and grafted WT animals showed no decline in their ability to sustain themselves on the rotating rod. In contrast, sham hSOD1(G93A) rats decreased in motor performance from week 16 onwards, reaching paralysis by week 19 of age. In grafted transgenic animals, there was a significant improvement in rotarod competence at weeks 16 and 17 when compared to sham hSOD1(G93A). However, in the following weeks, transplanted hSOD1(G93A) rats showed motor deterioration and eventually exhibited paralysis by week 19. At end-stage, we found only a few endogenous MN in sham and grafted hSOD1(G93A) rats by cresyl violet staining; no choline acetyl transferase-positive nor GFP-positive MN were present in grafted transgenic subjects. In contrast, WT rats analyzed at the same age possessed grafted GFP-positive MN in their spinal cords. These results strongly suggest that the transgenic hSOD1(G93A) environment is detrimental to grafted MN in the long term.