ESTROGENIC REGULATION OF GENE EXPRESSION DURING NEURONAL

近年来，雌激素受体(ER)在初级感觉中被发现。 大鼠背根神经节(DRG)的神经元，提示特异性 这些神经元的特性受性腺类固醇的调节 荷尔蒙。初步研究结果表明，几种主要的细胞骨架 蛋白质mRNAs和神经营养素受体mRNAs是靶标 背根神经节神经元的雌激素调节。这表明雌激素和雌激素受体 可能在调节神经突起生长特性中起重要作用。 背根神经节神经元。这个项目的长期目标是定义 雌激素对感觉神经元分子程序的影响 了解性腺类固醇如何影响卵巢的生长和再生 这些神经元。这项提案的具体目的是测试 以下是3个假设。第一个假设是内质网在卵巢中的表达 DRG受循环性腺类固醇激素的调节。就地 杂交，RNA印迹；免疫细胞化学将用于评估 雌激素治疗对雌激素受体和雌激素受体分布及水平的影响 其编码基因在去卵巢雌性大鼠背根神经节中的表达。 确定背根神经节ER/ER基因表达是否存在性别差异。 逆转录-聚合酶链式反应(RT-PCR)将用于 评估ER mRNA的选择性剪接作为激素状态的函数 和性别。第二个假设是雌激素调节 特定的细胞骨架和神经营养因子受体基因可以调节 背根节神经元的轴突切断反应。原位杂交、核糖核酸酶保护 检测和RNA印迹方法将被用来研究 雌激素对编码特异性mRNAs的稳态水平的影响 细胞骨架蛋白(BetaII、BetaIII和α-微管蛋白、外周蛋白和 Tau)和编码神经营养因子受体(p75、TrkA、TrkB)的基因 以及再生背根神经节神经元。第三个假设是 雌激素影响背根神经节神经元的再生特性。我们会 确定雌激素是否改变了最快的 生长的DRG轴突或再生较慢的人的生长特性 去卵巢雌性大鼠坐骨神经轴突的研究 使用快速轴突运输模式的周围神经挤压。性别 将在这些研究中评估再生参数的差异。 这项提议与健康的相关性在于潜在的识别 在神经元损伤方面的性别差异和新的信息中 将获得关于雌激素的潜在治疗作用的结论 神经元损伤。

Recently, estrogen receptors (ER) have been discovered in primary sensory neurons of the rat dorsal root ganglion (DRG), suggesting that specific properties of these neurons are subject to regulation by gonadal steroid hormones. Preliminary findings indicate that several major cytoskeletal protein mRNAs as well as neurotrophin receptor mRNAs are targets of estrogenic regulation in DRG neurons. This suggests that estrogen and ER may have an important role in regulating the neurite growth properties of DRG neurons. The long-term objectives of this project are to define the molecular programs affected by estrogen in sensory neurons and to understand how gonadal steroids influence the growth and regeneration of these neurons. The specific aims of this proposal are to test the following 3 hypotheses. The first hypothesis is that ER expression in the DRG is regulated by circulating gonadal steroid hormones. In situ hybridization, RNA blotting; immunocytochemistry will be used to evaluate the effects of estrogen treatment on the distribution and levels of ER and its encoding mRNA in the DRG of ovariectomized female rats, and to determine if sex differences in ER/ER mRNA expression in the DRG exist. Reverse transcription-polymerase chain reaction (RT-PCR) will be used to evaluate alternative splicing of ER mRNA as a function of hormone state and gender. The second hypothesis is that estrogenic regulation of specific cytoskeletal and neurotrophin receptor genes can modulate the axotomy response of DRG neurons. In situ hybridization, RNase protection assays and RNA blotting methods will be used to study the effects of estrogen on the steady-state levels of mRNAs encoding specific cytoskeletal proteins (BetaII, BetaIII and alpha1-tubulin, peripherin and tau) and those encoding neurotrophin receptors (p75, trkA, trkB) in normal as well as in regenerating DRG neurons. The third hypothesis is that estrogen influences the regenerative properties of DRG neurons. We will determine if estrogen alters the regeneration rate of either the fastest growing DRG axons or the growth properties of the more slowly regenerating axons in the sciatic nerve of ovariectomized female rats after a peripheral nerve crush using the fast axonal transport paradigm. Gender differences in regeneration parameters will be assessed in those studies. The health-relatedness of this proposal is in the potential identification of gender differences in neuronal injury and in the new information that will be gained concerning the potentially therapeutic role of estrogen in neuronal injury.