Creation and characterization of GH binding protein gene disrupted mice

GH 结合蛋白基因破坏小鼠的创建和表征

• 批准号: 7303768
• 负责人: John Joseph Kopchick
• 金额: $ 22.05万
• 依托单位: OHIO UNIVERSITY ATHENS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7303768
• 关键词: Aging; Alleles; Alternative Splicing; Animals; Binding; Biochemical; Biological; Biological Assay; Blood; Bone Development; Brain; Cell physiology; Communities; Complementary DNA; Complex; Condition; Deposition; Development; Diabetes Mellitus; Diabetic Nephropathy; Disease; Disruption; Endocrinology; Enhancers; Evolution; Excision; Exons; Extracellular Domain; Fatty acid glycerol esters; Fertility; Gene Targeting; Genes; Genomics; Goals; Growth; Growth Hormone Receptor; Growth and Development function; Hormones; Human; Insulin; Insulin Resistance; Kidney; Knock-out; Knockout Mice; Laboratories; Laron Syndrome; Left; Longevity; Malignant Neoplasms; Mammals; Mammary gland; Measurement; Mediating; Metabolic; Metabolism; Modeling; Molecular; Mus; Northern Blotting; Obesity; Oxidative Stress; Personal Satisfaction; Phenotype; Physiological; Play; Process; Production; Protein Binding; Proteins; Receptor Gene; Regulation; Relative (related person); Reproduction; Research; Rodent; Role; Scientist; Serum; Signal Transduction; Somatotropin; Somatropin; Structure of beta Cell of islet; Techniques; Temperature; Testing; Thinking; Tissues; Tooth structure; Western Blotting; blood glucose regulation; carbohydrate metabolism; diabetic; ghrelin; glucose metabolism; homologous recombination; human GHR protein; in vivo; inhibitor/antagonist; insight; knockout animal; mRNA Expression; mRNA Precursor; mouse growth hormone-binding protein; mouse model; prevent; protein function; receptor binding; reproductive; somatotropin-binding protein; theories; vector

DESCRIPTION (provided by applicant): To date, the biological role of growth hormone binding protein (GHBP) remains largely unknown. While many theories have been put forth, there is no direct evidence that can identify a specific role for this protein. We and others believe that it is highly likely that the GHBP plays a significant role in the GH/IGF- 1 axis. The main argument for this is due to the convergent evolution of two distinct mechanisms in mammals to produce the same protein (i.e. in humans the GHBP is produced by proteolytic cleavage of the GH receptor (GHR) extracellular domain while in rodents GHBP is produced by alternative splicing of the GHR precursor mRNA). Thus, we believe that the best way to elucidate the function of the GHBP is to create a GHBP gene specific disrupted or "knockout" mouse line. Hence, the objective of this research is to create the first GHBP specific gene-disrupted mouse and then to determine the contributions of GHBP on the growth phenotype as well as other metabolic processes in these animals. Our laboratory has already disrupted the mouse GHR/BP genes and generated combined GHR/GHBP "knockout" mice. In the homozygous state, these mice display a dwarf phenotype and serve as a model for a human GH- insensitive state termed Laron Syndrome. Although these mice are dwarf and obese, they are extremely sensitive to the action of insulin, resistant to diabetic kidney damage and, surprisingly, they have an extended life span. With this animal providing a background, we are now ready to test the specific hypotheses concerning the in vivo importance of GHBP by isolated GHBP gene-deficiency using a targeted gene disruption approach. We are confident that many questions concerning the biological role of GHBP will be answered in addition to new questions being raised once the GHBP gene disrupted mouse is generated. This proposal seeks to elucidate the role of growth hormone binding protein (GHBP), a protein whose function remains largely unknown other than its ability to bind growth hormone (GH) in the blood. Unlike GHBP, the function of GH is well established and GH is known to play an important role in normal metabolism and in the progression of several diseases such as diabetes, cancer and even aging. Thus, results obtained from this proposal would not only further our understanding of GH, but could help further our understanding of complex metabolic conditions and of the processes involved in aging.

描述（由申请人提供）：迄今为止，生长激素结合蛋白（GHBP）的生物学作用在很大程度上尚不清楚。尽管已经提出了许多理论，但没有直接的证据可以识别该蛋白质的特定作用。我们和其他人认为，GHBP很有可能在GH/IGF-1轴上发挥重要作用。这样做的主要论点是由于哺乳动物中两种不同机制的收敛演化（即在人类中，GHBP是通过GH受体（GHR）外细胞外结构域的蛋白水解裂解产生的，而在啮齿动物中GHBP则是由GHR Protursor MRNA的替代剪接产生的）。因此，我们认为阐明GHBP功能的最佳方法是创建GHBP基因特异性中断或“敲除”小鼠系。因此，这项研究的目的是创建第一个GHBP特异性基因中断的小鼠，然后确定GHBP对生长表型以及这些动物中其他代谢过程的贡献。我们的实验室已经破坏了小鼠GHR/BP基因，并产生了组合的GHR/GHBP“敲除”小鼠。在纯合状态下，这些小鼠表现出矮人表型，并作为称为拉隆综合征的人类GH-不敏感状态的模型。尽管这些小鼠是矮人和肥胖的，但它们对胰岛素的作用极为敏感，对糖尿病肾脏损害有抵抗力，而且令人惊讶的是，它们具有延长的寿命。通过这种动物提供背景，我们现在准备通过使用靶向基因破坏方法来测试通过孤立的GHBP基因缺陷来测试有关GHBP在体内重要性的特定假设。我们相信，除了生成GHBP基因中断的鼠标时提出的新问题外，还将回答有关GHBP生物学作用的许多问题。该提案旨在阐明生长激素结合蛋白（GHBP）的作用，该蛋白的功能在血液中结合生长激素（GH）的能力以外，其功能在很大程度上仍然未知。与GHBP不同，GH的功能已经确定，GH在正常代谢以及多种疾病（例如糖尿病，癌症甚至衰老）中起着重要作用。因此，从该提案中获得的结果不仅会进一步我们对GH的理解，还可以进一步帮助我们对复杂的代谢条件以及对衰老所涉及的过程的理解。