Novel Molecular Markers Governing Obesity in the Ventromedial Hypothalamus

描述（由申请人提供）： 在美国，儿童和青少年肥胖是一个严重的健康问题。在过去20年中，其流行率增加了10倍。2至18岁的儿童中有近15%现在超重（BMI>年龄和性别的第95百分位）。肥胖的增长趋势给我们的社会带来了巨大的健康和经济负担。控制肥胖的机制很复杂，尚未完全阐明。肥胖涉及无数环境和遗传因素之间的复杂相互作用。大脑中的下丘脑对调节和维持食物摄入和能量平衡至关重要。下丘脑腹内侧核（VMH）是下丘脑腹内侧核（VMH）的一个重要中枢。当啮齿类动物的VMH被破坏时，它们会变得贪食和肥胖。了解VMH功能的分子机制对肥胖的研究至关重要;然而，由于缺乏明确的VMH标志物，VMH的研究受到阻碍。迄今为止，类固醇生成因子（sf-1）是唯一的特异性标记VMH神经元，是关键的终末分化的VMH神经元。我们的实验室已经确定了一系列新的VMH富集基因与基因图谱和微阵列分析的使用。我的研究计划的总体目标是了解VMH在能量稳态中的生物学。该方法是研究新的VMH富集基因的功能。本研究的具体目标是：第一，通过评价一种新的转录因子前脑胚胎锌指（fez）的作用及其与sf-1的相互作用来研究VMH的发育;第二，通过研究囊泡谷氨酸转运体2（VGLUT 2）的作用及其在选择性地从VMH中去除时对食物摄入和体重的影响来研究VMH的兴奋功能;第三，通过鉴定在怀孕期间营养不良或营养过剩的雌性所生幼崽中差异表达的基因，来检查VMH的适应性和表观遗传功能。将使用稳定细胞系和正常、转基因和条件性敲除背景的小鼠进行研究。用于完成这些项目的研究技术将包括标准的分子生物学和遗传学技术，包括细胞培养，原位杂交和免疫细胞化学，转基因技术和微阵列分析。这项研究将有助于我作为一名独立的医生科学家的培训，以及确定管理儿童肥胖症的潜在治疗和预防方法。了解VMH的功能和机制将有助于我们了解下丘脑如何控制某些营养失调，如肥胖和II型糖尿病，以及其他复杂的人类行为，从神经性厌食症到性别焦虑症。

DESCRIPTION (provided by applicant): Childhood and adolescent obesity is a significant health problem in the United States. Its prevalence has increased 10 fold in the last 20 years. Almost 15% of children between 2 and 18 years old are now overweight (BMI > 95th percentile for age and gender). The increasing trend of obesity places a significant health and economic burden in our society. Mechanisms governing obesity are complex and not fully elucidated. Obesity involves a complex interaction among a myriad of environmental and genetic factors. The hypothalamus in the brain is critical to the regulation and maintenance of food intake and energy balance. Within the hypothalamus, a key center for satiety is the ventromedial hypothalamic nucleus (VMH). When VMH is destroyed in rodent species, they become hyperphagic and obese. Understanding the molecular mechanisms of VMH function is critical to the study of obesity; however, research in the VMH has been impeded by a lack of definitive VMH markers. To date, steroidogenic factor (sf-1) is the only specific marker for VMH neurons and is critical for the terminal differentiation of VMH neurons. Our laboratory has identified a list of novel VMH-enriched genes with the use of gene profiling and microarray analysis. The overall goal of my research proposal is to understand the biology of the VMH in energy homeostasis. The approach is to study the function of novel VMH-enriched genes. The specific aims are first, to study the development of the VMH by evaluating the role of a novel transcription factor, forebrain embryonic zinc-finger (fez), and its interaction with sf-1; second, to study the excitatory function of the VMH by examining the role of vesicular glutamate transporter 2 (VGLUT2) and its impact on food intake and body weight when it is selectively removed from the VMH; and third, to examine the adaptive and epigenetic function of the VMH by identifying genes that are differentially expressed in pups born to females that are undernourished or overnourished during pregnancy. The studies will be performed with stable cell lines and mice of normal, transgenic, and conditional knockout background. The research techniques used to accomplish these projects will include standard molecular biology and genetics techniques, including cell culture, in situ hybridization and immunocytochemistry, transgenic technologies, and microarray analysis. This research will facilitate my training as an independent physician scientist as well as identifying potential therapeutic and preventative approaches in managing childhood obesity. Understanding the function and mechanism of the VMH will help us to understand how the hypothalamus controls certain nutritional disorders, such as obesity, and type II diabetes, as well as other complex human behaviors, ranging from anorexia nervosa to gender dysphoria.