Transgenic resources for neuroscience research

用于神经科学研究的转基因资源

• 批准号: 8557119
• 负责人: James Pickel
• 金额: $ 211.96万
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8557119
• 关键词: Animals; Archives; Basic Science; Behavior; Biochemical; Biological Models; Callithrix; Cells; Collaborations; Complex; Core Facility; Development; Disease; Disease model; Embryo; Embryo Transfer; FOS Family Genes; Familial Dysautonomia; Female; Fright; Ganglioside Sialidase Deficiency Disease; Genes; Genetic; Genetic Research; Germ Cells; Goals; Harvest; Health Status; Hereditary Disease; Human; Imagery; Injection of therapeutic agent; Intramural Research Program; Ion Channel; Laboratories; Lead; Learning; Light; Link; Memory; Messenger RNA; Methods; Mus; National Institute of Drug Abuse; National Institute of Mental Health; Neurons; Neuropathy; Neurosciences; Neurosciences Research; Oocytes; Operative Surgical Procedures; Paper; Patients; Peptides; Pharmaceutical Preparations; Production; Protein Biosynthesis; RNA; Rattus; Recording of previous events; Research Personnel; Research Support; Resources; Rewards; Risk; Rodent; Role; Staging; Stress; Structure; Subfamily lentivirinae; Synapses; Techniques; Transcriptional Regulation; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Ultrasonography; United States National Institutes of Health; addiction; design; embryonic stem cell; experience; fascinate; human disease; mouse model; neural circuit; neural patterning; neurogenesis; neuropsychiatry; optogenetics; pathogen; promoter; recombinase; response; stem; tissue culture; trafficking; trait

SUMMARY The NIMH transgenic core facility has several major functions: 1) to produce transgenics for neuroscience research at NIH 2) develop new transgenic techniques and model systems 3) support research with associated techniques in genetic research in neuroscience and 4) engage in collaborative projects that promote genetic approaches to neuroscience research. 1) Production Meterics of production over the past year included: a) 23 transgenic mouse projects produced by oocyte injection, with multiple lines produced for each project. b) 14 transgenic rat projects produced by oocyte injection, with multiple lines produced for each project. c) 14 mouse projects have first altering the genes of ES cell and then using those to produce mice. 2) Development Over the last year projects to allow more productive methods have been developed. Two notable projects are a) in collaboration with NIDA the core has development of a panel of rats that produce recombinases or optogenetic probes under the control of transcriptional promoters that are specific for subpopulations of neurons. Fourteen of those lines have been produced, eight of which are being characterized by end users and will available for distribution. b) developing methods to produce transgenic marmosets. Over the past year rapid progress in developing techniques to produce transgenic marmosets have been made in the core facility. The methods to produce an excess of embryos, to optimize surgical procedures to harvest those embryos most efficiently, to culture those embryos, develop transgenic lentivirus, infect embryos, transfer those embryos into recipient females using ultrasound visualization and develop tissue culture methods for primary marmoset cells have been developed. This extensive effort has set the stage for several projects for NIH researchers to use these transgenic animals in basic research in the intramural program. 3) Technical Support a) 211 transgenic rodent lines have been archived by cryopreserving germ cells or embryos. b) 87 lines have been rederived, by transferring lines from pathogen bearing animals into those with defined health status. c) transgenic project design and assistance have continued to be significant to NIH neuroscience labs without experience in producing transgenic animals. 4) Collaborative projects: below is a list of projects that have been initiated in 2011, or have continued from last year. Stress and neurogenesis: Mice produced in the transgenic core were described in a paper that showed the necessity of neurogenesis for the normal response to stress. Since it has been shown that stress reduces neurogenesis this newer result indicates that a cycle of increasing stress. Learning and memory: The effect of specific and tightly controlled protein synthesis on learning and memory was studied. In addition, transgenic mouse models have been used to show the rrole of specific peptide-expressing cells to influence the link between fear and behavior and learning. Manipulating circuitry: Mice have been produced for two separate laboratories which have specific neurons that could be rendered transiently inactive by light activated ion channels. Those laboratories are investigating different neural circuits that are active in learning and addiction. Addictive and reward behavior: Lines of transgenic rats that express GFP in response to afferent input activation of the fos gene were generated in the core facility. These rats are being used by Bruce Hopes laboratory in NIDA to study patterns of neural activity in response to addictive drugs and most recently in the role of stress in reducing the re-establishment of rewarded behavior. . Mucolipidosis IV: The mouse model of this disease resulted from a long-standing collaboration with the Slaugenhaupt laboratory and has continued to yield results, including a description of the neuropathy that may be associated with this disease. The core facility continues to distribute these animals. More specific alterations of the Mcoln1 gene in transgenic animals are being planned. mRNA trafficking in neurons: An RNA stem loop structure is necessary for the translocation of message to specific cell compartments of the neuron. Mice that over express mRNA with this structure have been produced in an effort to disrupt this translocation machinery. By expressing this transgenic mRNA in different neuronal subtypes, the role for this mechanism for normal function is being studied. In addition this mechanism could be useful to target specific messages specifically to the synapse. Familial dysautonomia: Another collaboration with the Slaugenhaupt lab resulted in a model for this disease. Lines carrying either a human normal or disease gene are being created in the core. These lines are crossed into a null line to replace the endogenous IKBKAP gene with its human disease equivalent.

总结 NIMH转基因核心设施有几个主要功能：1）在NIH为神经科学研究生产转基因产品2）开发新的转基因技术和模型系统3）支持神经科学遗传研究中的相关技术研究和4）参与促进神经科学研究遗传方法的合作项目。 1)生产 过去一年的生产指标包括： a）23个通过卵母细胞注射产生的转基因小鼠项目，每个项目产生多个品系。 B）通过卵母细胞注射产生的14个转基因大鼠项目，每个项目产生多个品系。 c）14个小鼠项目首先改变ES细胞的基因，然后使用这些基因制造小鼠。 2)发展 在过去的一年里，已经制定了一些项目，以允许更有成效的方法。两个值得注意的项目是 a）与NIDA合作，核心开发了一组大鼠，其在对神经元亚群特异性的转录启动子的控制下产生重组酶或光遗传学探针。其中14条线路已经制作完成，8条线路正在由最终用户确定其特点，将可供分发。 B）开发生产转基因绒猴的方法。过去一年来，核心设施在开发生产转基因绒猴的技术方面取得了迅速进展。已经开发了产生过量胚胎的方法、优化外科手术以最有效地收获那些胚胎的方法、培养那些胚胎的方法、开发转基因慢病毒的方法、感染胚胎的方法、使用超声可视化将那些胚胎转移到受体雌性体内的方法以及开发用于原代绒猴细胞的组织培养方法。这种广泛的努力为NIH研究人员在校内项目的基础研究中使用这些转基因动物的几个项目奠定了基础。 3)技术支持 a）通过冷冻保存生殖细胞或胚胎，已存档了211个转基因啮齿动物系。 B）通过将品系从携带病原体的动物转移到具有确定的健康状态的动物中，已经重新衍生了87个品系。 c）转基因项目的设计和援助对于没有生产转基因动物经验的NIH神经科学实验室仍然很重要。 4)合作项目：以下是2011年启动的项目或去年继续开展的项目清单。 压力和神经发生：在一篇论文中描述了转基因核心中产生的小鼠，该论文显示了神经发生对于对压力的正常反应的必要性。由于已经表明压力会减少神经发生，这个新的结果表明压力的增加是一个循环。 学习和记忆：研究了特异性和严格控制的蛋白质合成对学习和记忆的影响。此外，转基因小鼠模型已被用于显示特定肽表达细胞的作用，以影响恐惧与行为和学习之间的联系。 操纵电路：已经为两个单独的实验室生产了小鼠，其具有可以通过光激活离子通道暂时失活的特定神经元。 这些实验室正在研究在学习和成瘾中活跃的不同神经回路。 成瘾和奖励行为：在核心设施中产生了响应于fos基因的传入输入激活而表达GFP的转基因大鼠系。NIDA的布鲁斯·霍普实验室正在使用这些老鼠来研究成瘾药物引起的神经活动模式，最近还研究了压力在减少奖励行为重建中的作用。 . 粘脂沉积症IV：这种疾病的小鼠模型来自与Slaugenhaupt实验室的长期合作，并继续产生结果，包括可能与这种疾病相关的神经病变的描述。核心设施继续分发这些动物。正在计划在转基因动物中对Mcoln 1基因进行更具体的改变。 神经元中的mRNA运输：RNA茎环结构对于将信息转运到神经元的特定细胞区室是必需的。为了破坏这种易位机制，已经产生了过表达具有这种结构的mRNA的小鼠。通过在不同的神经元亚型中表达这种转基因mRNA，正在研究这种机制对正常功能的作用。此外，这种机制可能有助于将特定的信息专门定向到突触。 家族性自主神经功能障碍：与Slaugenhaupt实验室的另一项合作导致了这种疾病的模型。携带人类正常或疾病基因的细胞系正在核心中产生。将这些系杂交成空系，以用其人类疾病等同物替换内源性IKBKAP基因。