Transgenic resources for neuroscience research

用于神经科学研究的转基因资源

• 批准号: 8342304
• 负责人: James Pickel
• 金额: $ 199.03万
• 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/8342304
• 关键词: Ablation; Adult; Age; Animal Model; Animals; Anxiety; Archives; Basic Science; Behavior; Behavioral; Binding; Biochemical; Biological Models; Brain; Calcium; Callithrix; Cells; Characteristics; Collaborations; Complex; Core Facility; Development; Disease; Disease model; Drug Addiction; Embryo; Embryo Transfer; Engineering; Enterobacteria phage P1 Cre recombinase; FOS Family Genes; Familial Dysautonomia; Family; Female; Fright; Functional disorder; Ganglioside Sialidase Deficiency Disease; Genes; Genetic; Genetic Research; Germ Cells; Goals; Harvest; Health Status; Hereditary Disease; Human; Imagery; Injection of therapeutic agent; Interneurons; Intramural Research Program; Ion Channel; Laboratories; Lead; Learning; Light; Link; Memory; Methods; Methyltransferase; Mus; N-Methyl-D-Aspartate Receptors; National Institute of Drug Abuse; National Institute of Mental Health; Neurodegenerative Disorders; Neuroglia; Neurons; Neuropathy; Neurosciences; Neurosciences Research; Oocytes; Operative Surgical Procedures; Paper; Partner in relationship; Patients; Peptides; Pharmaceutical Preparations; Predisposition; Pregnancy; Production; Property; Protein Biosynthesis; Rattus; Recording of previous events; Reporter; Research Personnel; Research Support; Resources; Risk; Rodent; Role; Schizophrenia; Societies; Staging; Stress; Subfamily lentivirinae; Techniques; Transcriptional Regulation; Transgenes; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Ultrasonography; United States National Institutes of Health; Work; addiction; design; embryonic stem cell; experience; human disease; interest; mature animal; mouse model; neural circuit; neural patterning; neurogenesis; neuropsychiatry; offspring; pathogen; promoter; response; tissue culture; 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) 48 transgenic mouse lines produced by oocyte injection. b) 14 rat lines have been produced by oocyte injection. c) 13 mouse projects have first altering the genes of embryonic stem 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) development of a panel of rats that produce CRE recombinase under the control of transcriptional promoters that are specific for subpopulations of neurons. 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) 98 transgenic rodent lines have been archived by cryopreserving germ cells or embryos. b) 68 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: The role of a specific gene (catachol-O-methyltransferase) in the susceptibility to stress was demonstrated in mice that were engineered to have reduced levels of this gene. Neurogenesis: Transgenic mice and, more recently, transgenic rats have been generated to study the role of neurogenesis in adults. From mid-gestation and continuing into old age, new neurons are produced in the brain. The role of new cells appearing in adults is especially interesting, and suggests a function in learning and memory and potential treatments for neurodegenerative disease. Schizophrenia: Mice with behavioral characteristics that resemble schizophrenia were produced by mating effecter mice from the core facility with responder mice. The latter carried a conditional ablation of the NMDA receptor. The offspring of these matings were engineered to lack NMDA receptors in a subset of corticolimbic interneurons. They displayed behavioral deficits in normal mating, nest-building and anxiety-like behavior. Memory dysfunction was also revealed in these animals. Stress and neurogenesis: A recent paper using mice developed in the transgenic core has shown that neurogenesis is critical for maintaining a normal response to stress. This finding is significant because it conlusively identifies this role for neurogenesis in adult animals. Following other work that shows that stress reduces neurogenesis, this new finding implies a cycle in which stress can increase. 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 role 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. Drug addiction: 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. . 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. 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. Glial activity reporters: The core produced a mouse line with a transgene that indicated the concentration of calcium in glial cells. By changes in its fluorescent properties, the calcium concentration and associated activity has been demonstrated in these cell. Reporter and effector mice and rats:

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）通过卵母细胞注射产生的48个转基因小鼠系。 B）通过卵母细胞注射产生了14个大鼠品系。 c）13个小鼠项目首先改变胚胎干细胞的基因，然后用这些基因来制造小鼠。 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）开发一组大鼠，其在对神经元亚群特异的转录启动子的控制下产生CRE重组酶。 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，已将98个转基因啮齿动物品系存档。 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68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：一个特定的基因（catachol-O-甲基转移酶）在对压力的易感性中的作用在被改造成具有降低的该基因水平的小鼠中得到了证明。 神经发生：转基因小鼠和最近的转基因大鼠已经被用来研究成年人神经发生的作用。从妊娠中期到老年，大脑中产生新的神经元。成年人中出现的新细胞的作用特别有趣，表明其在学习和记忆中的功能以及神经退行性疾病的潜在治疗方法。 精神分裂症：具有类似精神分裂症的行为特征的小鼠是通过将来自核心设施的效应小鼠与应答小鼠交配产生的。后者携带NMDA受体的条件性消融。这些交配的后代经过设计，在皮质边缘中间神经元的一个子集中缺乏NMDA受体。它们在正常交配、筑巢和焦虑行为方面表现出行为缺陷。在这些动物中也发现了记忆功能障碍。 压力和神经发生：最近一篇使用转基因核心小鼠的论文表明，神经发生对于维持对压力的正常反应至关重要。这一发现意义重大，因为它最终确定了成年动物神经发生的作用。在其他研究表明压力会减少神经发生之后，这一新发现暗示了压力会增加的循环。 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。 药物成瘾：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对成瘾药物反应的神经活动模式。 . 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。 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。 神经胶质活性报告基因：核心产生了一个转基因小鼠品系，该转基因指示神经胶质细胞中的钙浓度。通过其荧光特性的变化，已经证明了这些细胞中的钙浓度和相关活性。 报告子和效应子小鼠和大鼠：