ETHANOL CEREBELLAR PATHOLOGY--RETINOIDS AND GLYCOLIPIDS

描述：(改编自《调查者摘要》)小脑病理 胎儿酒精综合征(FAS)是酒精暴露的一种有害影响 人类和大鼠的发育。我们的体内数据显示，乙醇给药 胃内增加小脑中维甲酸(RA)的水平，维甲酸是一种有效的 形成剂。此外，我们的小脑颗粒培养的体外数据 细胞，小脑星形胶质细胞，或两者的共同培养表明，乙醇 外源RA减少岩藻糖化阶段特异性产物的产生 胚胎抗原-1(SSEA-1)和9-O-乙酰神经节苷脂(Jones) 被认为是细胞黏附和迁移的基本糖偶联物 沿着发育中的小脑的神经胶质突起。此外，乙醇和 这些培养物中外源RA降低岩藻糖基转移酶mRNA的表达， 可能是SSEA-1生物合成途径中的速率决定酶。这些 数据表明，酒精神经病理与小脑RA的增加有关 改变细胞表面对细胞至关重要的糖结合物表达的水平 黏附、迁徙和生存。这一点得到了 外源性维甲酸对出生后大鼠延缓死亡的观察 阻止小脑颗粒细胞迁移和/或存活，以及SSEA-1 在体外，特异性抗体可阻止颗粒细胞与神经胶质细胞的黏附。 我们的假设是，酒精小脑病理与RA的增加有关 这会减少细胞表面糖结合物的表达。我们的具体目标 测定：1)小脑RA及其合成物的出生后水平 乙醇或维甲酸对新生大鼠体内酶和受体的影响 9-O-乙酰神经节苷脂、GD3-9-O-乙酰基转移酶的出生后水平 活性、SSEA-1糖结合物、岩藻糖基转移酶活性、 岩藻糖基转移酶mRNA，并评价浦肯野细胞存活、颗粒细胞 出生后大鼠体内治疗后的存活/迁移和胶质细胞含量 乙醇或RA；3)RA水平、合成酶和RA的表达 乙醇对培养的小脑颗粒细胞、星形胶质细胞受体的影响 细胞，以及两种细胞类型的共培养；4)细胞存活的变化 以及这些细胞在外源性RA或抗体存在时的细胞黏附 SSEA-1或9-O-乙酰神经节苷脂的特异性；5) 9-O-乙酰神经节苷脂、SSEA-1糖结合物、岩藻糖基转移酶活性和 乙醇或RA暴露的这些细胞培养中岩藻糖基转移酶基因的表达。

DESCRIPTION: (Adapted from the Investigator's Abstract) Cerebellar pathology in fetal alcohol syndrome (FAS) is a detrimental effect of ethanol exposure during development in humans and rats. Our in vivo data show that ethanol administered intragastrically increases cerebellar levels of retinoic acid (RA), a potent morphogen. Further, our in vitro data from cultures of cerebellar granule cells, cerebellar astrocytes, or co-cultures of the two show that both ethanol and exogenous RA decrease the production of fucosylated stage-specific embryonic antigen-1 (SSEA-1) and 9-O-acetyl gangliosides (JONES); antigens thought to be essential glycoconjugates for cell adhesion to and migration along glial processes in the developing cerebellum. Further, both ethanol and exogenous RA in these cultures decrease expression of fucosyltransferase mRNA, likely the rate-determining enzyme in the SSEA-1 biosynthetic pathway. These data imply that ethanol neuropathology involves an increase in cerebellar RA levels which alter cell surface expression of glycoconjugates critical to cell adhesion, migration, and survival. This is further supported by the observations that exogenous RA administrated to postnatal rats delays or prevents cerebellar granule cell migration and/or survival and that SSEA-1 specific antibodies prevent granule cell adhesion to glial processes in vitro. Our hypothesis is that ethanol cerebellar pathology involves an increase in RA which decreases cell surface expression of glycoconjugates. Our specific aims are to determine: 1) The postnatal levels of cerebellar RA and its synthetic enzymes, and receptors in neonatal rats treated in vivo with ethanol or RA; 2) The postnatal levels of 9-O-acetyl gangliosides, GD3-9-O-acetyltransferase activity, SSEA-1 glycoconjugates, fucosyltransferase activity, fucosyltransferase mRNA, and to evaluate Purkinje cell survival, granule cell survival/migration, and glial cell content in postnatal rats treated in vivo with ethanol or RA; 3) The RA levels, synthetic enzymes, and expression of RA receptors in ethanol-exposed cultures of cerebellar granule cells, astroglial cells, and co-cultures of the two cell types; 4) Alterations in cell survival and cell adhesion of these cells in the presence of exogenous RA, or antibodies specific to SSEA-1 or 9-O-acetyl gangliosides; 5) The content and expression of 9-O-acetylgangliosides, SSEA-1 glycoconjugates, fucosyltransferase activity and fucosyltransferase mRNA in ethanol- or RA-exposed cultures of these cells.