Genetic and cellular sex differences in cerebellum

描述(由申请人提供)：在许多神经-运动疾病中，强烈的性别差异被很好地记录下来，然而，关于运动系统的实验工作很少包括性别，当它包括性别时，没有进行性别比较。这一差距是严重的，将通过本提案中的工作加以解决。小脑是大脑的主要协调中枢，它的失调会导致多种运动性疾病。这项研究的最终目标是确定性染色体基因突变是否在性二型神经运动障碍中发挥关键作用。这一建议的基础是观察到雌性小鼠或具有XX性染色体补充的雌性小鼠比雄性小鼠或任何XY个体在小脑中有更多的钙结合蛋白表达。Calbindin D28K仅存在于小脑中唯一的投射神经元--浦肯野细胞。这种蛋白质与大脑中的神经保护有关，水平降低与许多神经疾病有关。该项目的最初目标是确定性别差异是否是浦肯野细胞数量差异的结果。下一个问题是确定浦肯野细胞中是否存在性别差异的树突结构。最后，我们将确定特定的SUB 小脑的部分区域比其他区域更具性别二型性。这些目标将通过组织学染色和形态计量学分析来实现，此外还将进行蛋白质印迹和Q RT-PCR。我们将在小鼠品系中进行所有研究，允许对性别、染色体组成和性腺性别(即卵巢和睾丸)进行单独分析。接下来，我们将揭示导致性别差异的基因。这将使用一组工程小鼠模型来完成，这些模型能够通过基因表达微阵列对细胞特异性的mRNA进行量化。我们的假设是，性染色体基因(S)调节钙结合蛋白，这两个因素一起以性别二态的方式改变浦肯野细胞的连接。这一计划提供了一个基于性染色体基因的关于小脑功能性别差异的新视角；这将导致神经运动性疾病患者的诊断和干预方面的创新。

DESCRIPTION (provided by applicant): Strong gender differences are well documented for many neural-motor diseases, yet, experimental work on motor systems rarely includes both genders and when it does, sex comparisons are not made. This gap is critical and will be addressed by the work in this proposal. The cerebellum is the master coordination center in the brain and its dysregulation produces many kinds of motor diseases. The ultimate goal of this research is to determine if mutations in sex chromosome genes play key roles in sexually dimorphic neurological motor disorders. The basis for the proposal is the observation that female mice, or mice of either sex with an XX sex chromosome complement, have more calbindin expression in the cerebellum than males or any XY individuals. Calbindin D28K is only present in the Purkinje cells, the only projection neurons in the cerebellum. This protein is linke with neuroprotection in the brain and reduced levels have been associated with many neurological diseases. The initial goal of this project is to determine if sex differences are the result of differences in numbers of Purkinje cells. The next question is to determine if there are sex differences dendritic structures in Purkinje cells. Finally we will determine if particular sub regions of the cerebellum are more sexually dimorphic than others. These goals will be accomplished with histological staining and morphometric analyses in addition to western blots and q RT-PCR. We will do all the studies in a mouse line allows separate analysis of sex chromosome complement and gonadal sex (i.e. ovaries and testes). Next, the genes that contribute to the sex differences will be revealed. This will be accomplished using a combination of engineered mouse models that enable cell-specific quantification of mRNA via gene expression microarray. Our hypothesis is that a sex chromosome gene(s) regulate calbindin and together these two factors modify Purkinje cell connections in a sexually dimorphic manner. This program offers a novel perspective on sex differences in cerebellar function based on sex chromosome genes; this will lead to innovations in diagnostics and interventions for patients with neurological motor diseases.