Cell Biiology Core

核心B细胞生物学核心（核心B）的目的是通过提供特定年龄小鼠的睾丸、卵巢和富集的生殖细胞来实现该计划的目标，以支持项目B Petkov、C Hibbs和D Handel中的ChIP-seq实验;项目C Hibbs和D Handel中的RNA-seq实验;以及项目D Handel中的RNA和蛋白质表达分析。核心B将利用严格控制的方法从幼年睾丸中获得生殖细胞，用于遵循精子发生高度同步化的第一波前沿的试验。此外，核心B将使用单位重力下的细胞沉降技术，以富集来自成年睾丸的粗线期精母细胞和圆形精子。核心将为项目中提出的实验执行RNA纯化和染色质免疫沉淀的初始步骤。核心B将在Handel博士的指导下进行，Handel博士在减数分裂和雄性生殖细胞生物学研究方面经验丰富，并具有分离小鼠精母细胞的专业知识。总的来说，核心B的功能确保所有项目都将拥有高质量和一致的实验材料，以实现单个项目目标以及共同实现计划的总体目标。155细胞生物学核心（Handel）Paigen，Kenneth具体目标细胞生物学核心（核心B）将支持该计划的细胞和分子需求。这一目标将确保所有项目的生殖细胞和生殖细胞提取物的高质量和一致性，以便该计划的实验结果具有直接可比性。此外，在经验丰富的领导者的指导下，将这些职能集中到一个核心中，可以提高成本和职能的效率。核心将为利用性腺或生殖细胞的实验提供材料，包括表达分析（项目D Handel）、RNA-seq（项目C Hibbs和D Handel）和ChlP-seq（项目B Petkov、C Hibbs和D Handel）。细胞生物学核心的目标是：具体目标1。核心将从小鼠中获取性腺，并从幼龄和成熟小鼠中制备富集的生殖细胞，用于该计划的实验。具体目标2。核心将从性腺和/或生殖细胞中制备RNA和蛋白质提取物，用于该计划的表达分析和RNA-seq实验。具体目标3。核心将对性腺或生殖细胞进行染色质免疫沉淀，以支持该计划的ChIP-seq实验。总的来说，这些目标确保所有研究人员都将拥有必要的实验材料，以实现他们的个人项目目标以及共同实现该计划的总体目标。

CORE B The objective of the Cell Biology Core (Core B) is to enable goals of the Program by providing testes, ovaries and enriched germ cells from mice of specific ages to support ChlP-seq experiments in Projects B Petkov, C Hibbs and D Handel; RNA-seq experiments in Projects C Hibbs and D Handel; and RNA and protein expression analyses in Project D Handel. Core B will utilize carefully controlled methods to obtain germ cells from juvenile testes for assays that follow the leading edge of the highly synchronized first wave of spermatogenesis. Additionally, Core B will use a technique of cell sedimentation at unit gravity to enrich pachytene spermatocytes and round sperm fids from adult testes. The Core will perform initial steps of RNA purification and chromatin-immunoprecipitation for experiments proposed in the Projects. Core B will be under the direction of Dr. Handel, who is highly experienced in investigation of meiosis and male germ-cell biology, and has expertise in isolation of mouse spermatocytes. Taken together, the functions of Core B ensure that all Projects will have high-quality and consistent experimental materials to achieve the individual Project goals as well as to collectively achieve the overall goals of the Program. 155 Cell Biology Core (Handel) Paigen, Kenneth SPECIFIC AIMS The Cell Biology Core (Core B) will support cellular and molecular needs of the Program. This goal will ensure high-quality and consistent preparations of germ cells and germ-cell extracts across all Projects, so that experimental results in the Program will be directly comparable. Moreover, centralizing these functions into a Core, under the direction of an experienced leader, provides efficiencies of cost and function. The Core will provide material for experiments that utilize gonads or germ cells, including expression analyses (Project D Handel), RNA-seq (Projects C Hibbs and D Handel), and ChlP-seq (Projects B Petkov, C Hibbs and D Handel). The aims for the Cell Biology Core are: Specific Aim 1. The Core will retrieve gonads from mice and prepare enriched germ cells from juvenile and mature mice to be used in experiments of the Program. Specific Aim 2. The Core will prepare extracts of RNA and protein from gonads and/or germ cells to be used for expression analyses and RNA-seq experiments of the Program. Specific Aim 3. The Core will conduct chromatin-immunoprecipitation on gonads or germ cells to support ChlP-seq experiments of the Program. Overall, these aims ensure that all investigators will have the necessary experimental materials to achieve their individual Project goals as well as to collectively achieve the overall goals of the Program.