ETHANOL CEREBELLAR PATHOLOGY--RETINOIDS AND GLYCOLIPIDS

乙醇小脑病理学——类维生素A和糖脂

• 批准号: 6132916
• 负责人: M D ULLMAN
• 金额: $ 0.85万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6132916
• 关键词: cerebellum; developmental neurobiology; ethanol; fetal alcohol syndrome; glycolipids; laboratory rat; neuropathology; newborn animals; retinoids

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.

描述：（改编自研究者摘要） 胎儿酒精综合征（FAS）是一种有害的影响，乙醇暴露期间， 在人类和老鼠的发展。我们的体内数据显示， 灌胃增加小脑维甲酸（RA）水平， 形态发生素此外，我们来自小脑颗粒培养物的体外数据 细胞，小脑星形胶质细胞，或两者的共培养物显示，乙醇 和外源性RA减少岩藻糖基化阶段特异性 胚胎抗原-1（SSEA-1）和9-O-乙酰神经节苷脂（JONES）;抗原 被认为是细胞粘附和迁移所必需的糖缀合物 沿着神经胶质突起。此外，乙醇和 这些培养物中的外源性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，乙醇神经病理学涉及小脑RA的增加， 改变对细胞关键的糖缀合物的细胞表面表达水平 粘附、迁移和存活。这一点得到了 观察到外源性RA给予出生后大鼠延迟或 阻止小脑颗粒细胞迁移和/或存活， 特异性抗体在体外阻止颗粒细胞粘附到神经胶质突起。 我们的假设是，乙醇小脑病理涉及增加类风湿关节炎 其降低糖缀合物的细胞表面表达。我们的具体目标 本研究旨在测定：1）小脑RA及其合成代谢产物的生后水平 酶和受体在体内用乙醇或RA处理的新生大鼠; 2） 9-O-乙酰神经节苷脂、GD 3 -9-O-乙酰转移酶的生后水平 活性，SSEA-1糖缀合物，岩藻糖基转移酶活性， 岩藻糖基转移酶mRNA，并评估浦肯野细胞存活、颗粒细胞 存活/迁移和胶质细胞含量 3）RA水平、合成酶和RA表达 小脑颗粒细胞、星形胶质细胞和神经胶质细胞的乙醇暴露培养物中的受体 细胞和两种细胞类型的共培养物; 4）细胞存活的改变 以及在外源性RA或抗体存在下这些细胞的细胞粘附 特异于SSEA-1或9-O-乙酰基神经节苷脂; 5） 9-O-乙酰神经节苷脂、SSEA-1糖缀合物、岩藻糖基转移酶活性和 岩藻糖基转移酶mRNA在这些细胞的乙醇或RA暴露培养物中的表达。