Genetic and cellular sex differences in cerebellum

描述（由申请人提供）：许多神经运动疾病都有很强的性别差异，然而，运动系统的实验工作很少包括两种性别，即使这样，也没有进行性别比较。这一差距至关重要，将通过本提案中的工作加以解决。小脑是大脑中的主要协调中心，其失调会导致多种运动疾病。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态性神经运动障碍中起关键作用。该提议的基础是观察到雌性小鼠或具有XX性染色体互补的任何性别的小鼠在小脑中比雄性或任何XY个体具有更多的钙结合蛋白表达。钙结合蛋白D28 K仅存在于浦肯野细胞中，浦肯野细胞是小脑中唯一的投射神经元。这种蛋白质与大脑中的神经保护有关，并且水平降低与许多神经系统疾病有关。这个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的树突结构是否存在性别差异。最后，我们将确定特定的子 小脑的区域比其他区域更具有性别二态性。除蛋白质印迹和qRT-PCR外，还将通过组织学染色和形态测定分析实现这些目标。我们将在允许单独分析性染色体补体和性腺性别（即卵巢和睾丸）的小鼠品系中进行所有研究。接下来，将揭示导致性别差异的基因。这将使用工程小鼠模型的组合来实现，所述工程小鼠模型能够通过基因表达微阵列对mRNA进行细胞特异性定量。我们的假设是，性染色体基因（S）调节钙结合蛋白，这两个因素一起修改浦肯野细胞连接的性二态性的方式。该计划提供了基于性染色体基因的小脑功能性别差异的新视角;这将导致神经运动疾病患者诊断和干预的创新。

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.