Sphingolipid Biosynthesis Is Necessary for Dendrite Growth and Survival of Cerebellar Purkinje Cells in Culture
Sphingolipid Biosynthesis Is Necessary for Dendrite Growth and Survival of Cerebellar Purkinje Cells in Culture
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鞘脂生物合成对于培养中小脑浦肯野细胞的树突生长和存活是必要的
DOI：
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发表时间：
1995
影响因子：
4.7
通讯作者：
Y. Hirabayashi
中科院分区：
文献类型：
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作者：
S. Furuya;K. Ono;Y. Hirabayashi
摘要：在发育中的大鼠小脑浦肯野神经元中，使用分离的培养系统研究了神经元生长对复合鞘脂生物合成的需求。浦肯野细胞在无血清营养条件下2周后发育出分化良好的树突和轴突。添加2 µM伏马菌素B1（一种哺乳动物神经酰胺合成酶的真菌抑制剂）可抑制[3 H]半乳糖/葡糖胺和[14 C]丝氨酸掺入培养的小脑神经元的复合鞘脂中。在此条件下，浦肯野细胞富集鞘脂（包括GD 1 α、9-O-乙酰化LD 1和GD 3）和鞘磷脂的表达显著降低。暴露于伏马菌素B1 2周后，在伏马菌素处理的浦肯野细胞中观察到剂量依赖性可测量的存活率降低和目视可辨别的形态学差异。浦肯野细胞树突表现出两种类型的异常;一个细胞群体在轻微的时间滞后后发育出细长但分支较少的树突，但它们的分支开始退化。在一些细胞中，细长的树突树的形成严重受损。然而，伏马菌素B1的治疗也导致浦肯野细胞树突上的棘状突起的形成，在控制文化。与在浦肯野细胞中观察到的改变相反，用伏马菌素B1处理后，包括颗粒神经元在内的其他细胞类型的形态似乎几乎正常。这些观察结果强烈表明，膜鞘脂参与树突的生长和维持，并在小脑浦肯野细胞的生存。事实上，伏马菌素B1的这些作用通过添加神经酰胺的合成衍生物6-[[N-（7-硝基苯并-2-氧杂-1，3-二唑-4-基）-氨基]己酰基]鞘氨醇（C6-NBD-神经酰胺）而逆转，但不完全逆转。因此，我们得出结论，剥夺膜鞘脂在培养环境中是负责浦肯野细胞的异常生长。
Abstract: The requirement of complex sphingolipid biosynthesis for growth of neurons was examined in developing rat cerebellar Purkinje neurons using a dissociated culture system. Purkinje cells developed well‐differentiated dendrites and axons after 2 weeks in a serum‐free nutrient condition. Addition of 2 µM fumonisin B1, a fungal inhibitor of mammalian ceramide synthase, inhibited incorporation of [3H]galactose/glucosamine and [14C]serine into complex sphingolipids of cultured cerebellar neurons. Under this condition, the expression of Purkinje cell‐enriched sphingolipids, including GD1α, 9‐O‐acetylated LD1 and GD3, and sphingomyelin, was significantly decreased. After 2 weeks' exposure to fumonisin B1, dose‐dependent measurable decreases in the survival and visually discernible differences in the morphology were seen in fumonisin‐treated Purkinje cells. The Purkinje cell dendrites exhibited two types of anomalies; one population of cells developed elongated but less‐branched dendrites after a slight time lag, but their branches began to degenerate. In some cells, formation of elongated dendrite trees was severely impaired. However, treatment with fumonisin B1 also led to the formation of spinelike protrusions on the dendrites of Purkinje cells as in control cultures. In contrast to the alterations observed in Purkinje cells, morphology of other cell types including granule neurons appeared to be almost normal after treatment with fumonisin B1. These observations indicated strongly that membrane sphingolipids participate in growth and maintenance of dendrites and in the survival of cerebellar Purkinje cells. Indeed, these effects of fumonisin B1 were reversed, but not completely, by the addition of 6‐[[N‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)‐amino]caproyl]sphingosine (C6‐NBD‐ceramide), a synthetic derivative of ceramide. Thus, we conclude that deprivation of membrane sphingolipids in a culture environment is responsible for aberrant growth of Purkinje cells.
神经酰胺的荧光衍生物：物理性质及其在研究动物细胞高尔基体中的用途。
影响因子：
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作者：
Pagano,RE
通讯作者：
Pagano,RE