Guigoni Céline, Coulon Patrice
Groupe d'Etude des Réseaux Moteurs, CNRS, Marseille, France.
J Neurovirol. 2002 Aug;8(4):306-17. doi: 10.1080/13550280290100761.
Cultures of purified rat embryonic spinal cord motoneurons were used to investigate the capacity of the neurons to survive rabies virus infection in vitro. In crude primary spinal cord cultures, neurons did not survive more than 2 days after rabies virus infection with the fixed strain Challenge Virus Standard. In contrast, virus-infected purified motoneurons resisted cytolysis for at least 7 days, as also did infected motoneurons treated with conditioned medium sampled from rabies virus-infected crude spinal cord cultures. This survival rate was also observed when motoneurons were grown in the presence of astrocytes or fibroblasts and it was not dependent on the presence of growth factors in the culture medium. Moreover, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling experiments showed that only 30% of infected motoneurons were apoptotic after 7 days of infection. In vivo, despite the massive infection of the spinal cord in infected rat neonates, the moderate number of apoptotic cells in the ventral horn suggests that only a few motoneurons were affected by this mechanism of cell death. Morphometric analyses showed that motoneurons' axon elongated at a comparable rate in virus-infected and noninfected cultures, a sign of high metabolic activity maintained in rabies virus-infected motoneurons. In contrast, hippocampus neurons were susceptible to rabies virus infection, because 70% of infected neurons were destroyed within 3 days, a large proportion of them being apoptotic. These experiments suggest that spinal cord motoneurons consist in a neuronal population that survive rabies virus infection because the viral induction of apoptosis is delayed in these neurons. They suggest also that paralyses frequently observed in rabid animals could be the consequence of dysfunctions of the locomotor network or of the spinal cord motoneurons themselves, whose parameters could be studied in vitro.
使用纯化的大鼠胚胎脊髓运动神经元培养物来研究这些神经元在体外抵抗狂犬病病毒感染的能力。在粗制的原代脊髓培养物中,用固定毒株攻击病毒标准株感染狂犬病病毒后,神经元存活不超过2天。相比之下,病毒感染的纯化运动神经元至少7天抵抗细胞溶解,从狂犬病病毒感染的粗制脊髓培养物中采集的条件培养基处理的感染运动神经元也是如此。当运动神经元在星形胶质细胞或成纤维细胞存在的情况下生长时,也观察到了这种存活率,并且它不依赖于培养基中生长因子的存在。此外,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记实验表明,感染7天后只有30%的感染运动神经元发生凋亡。在体内,尽管感染的新生大鼠脊髓受到大量感染,但腹角中凋亡细胞数量适中,这表明只有少数运动神经元受这种细胞死亡机制的影响。形态计量分析表明,病毒感染和未感染的培养物中运动神经元的轴突以相当的速度伸长,这是狂犬病病毒感染的运动神经元中维持高代谢活性的一个迹象。相比之下,海马神经元易受狂犬病病毒感染,因为70%的感染神经元在3天内被破坏,其中很大一部分是凋亡的。这些实验表明,脊髓运动神经元是一群能在狂犬病病毒感染中存活的神经元,因为这些神经元中病毒诱导的凋亡被延迟。它们还表明,狂犬病动物中经常观察到的麻痹可能是运动网络或脊髓运动神经元本身功能障碍的结果,其参数可以在体外进行研究。