ALTERED ETHANOL EFFECTS ON OSTEOCALCIN NULL MUTANT MICE

改变乙醇对骨钙素无效突变小鼠的影响

• 批准号: 6873112
• 负责人: PATRICIA E PATTERSON-BUCKENDAHL
• 金额: $ 19.74万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6873112
• 关键词: alcoholic beverage consumption; behavioral /social science research tag; behavioral genetics; binding proteins; calcium metabolism; ethanol; genetically modified animals; hormone regulation /control mechanism; immunocytochemistry; laboratory mouse; microinjections; neural transmission; neuroanatomy; osteoblasts; osteocalcin; preference; psychological stressor; southern blotting; stress

Preliminary data showed that preference for and consumption of alcohol (EtOH) by osteocalcin (OC) null mutant mice (KO) differs significantly from wildtype C57BL/6 (WT) mice. Their behavior both in terms of basic behaviors and in response to EtOH administration also differed. Circulating osteocalcin (pOC), a protein of bone origin which generally reflects osteoblastic activity, may be decreased or increased following EtOH consumption. Variation in EtOH intake depends on dose, age; gender, and duration of alcohol consumption. Furthermore, pOC is strongly affected by stressful stimuli, again varying with the type and duration of the stressor and the elapsed time following stress exposure, pOC is rapidly increased within 3 to 5 minutes of exposure to acute severe stressors and returns toward basal levels after 1 to 2 hours. Milder, anxiety producing stressors slowly decrease pOC over 1 to 2 hours. It is also accepted that stressful experiences tend to increase alcohol consumption. There have been reports of immunolocalization of OC protein in sympathetic ganglia of rats, and of expression of OC mRNA in brain of rats and mice. Experimental approach: Research proposed here will (1) evaluate alcohol consumption and preference in KO and WT mice, (2) determine effect of acute supplementation of OC on EtOH drinking in KO mice, (3) evaluate EtOH drinking. and hormonal responses to restraint stress, and (4) probe for presence of OC by immunolocalization in extraskeletal tissues, including sympathetic ganglia, spinal cord, and brain. Significance: To date: pOC has been used primarily as a biomarker of bone formation, yet it is consistently altered by EtOH ingestion and by various psychological stressors in ways that cannot be attributed to osteoblast activity. The literature on the association of EtOH ingestion and stress has been somewhat controversial. The proposed experiments are expected to provide at least one potential answer to this controversial field. The hormonal changes that affect ingestion of EtOH in response to a severe stressor may differ in KO mice. Thus we expect to provide initial evidence for a novel mechanism for stress associated changes in EtOH ingestion. Positive identification by immunocytochemistry of OC interaction with various neural locations will provide essential information regarding its potential site of action in regulating alcohol ingestion. Furthermore, the proposed experiments will demonstrate that pOC is not a passive component of plasma, but rather part of a feedback loop emanating from bone to regulate neural input involving EtOH consumption and response to stressful conditions.

初步数据表明，骨钙素（OC）无效突变小鼠（KO）对酒精（ETOH）的偏爱与WildType C57BL/6（WT）小鼠明显不同。他们的行为在基本行为方面和对ETOH给药的响应也有所不同。循环骨钙素（POC）是一种通常反映成骨细胞活性的骨骼来源的蛋白质，在ETOH消耗后可能会降低或增加。 EtOH摄入量的变化取决于剂量，年龄；性别和饮酒持续时间。此外，POC受到压力刺激的强烈影响，再次随压力源的类型和持续时间而变化，压力暴露后经过的时间变化，POC在暴露于急性严重压力源的3至5分钟内迅速增加，并在1至2小时后返回基础水平。温和的，产生焦虑的压力慢慢降低了1至2小时的POC。还接受压力的经历倾向于增加酒精消耗。有报道称，OC蛋白在大鼠交感神经节中的免疫定位以及大鼠和小鼠大脑中OC mRNA的表达。 实验方法：此处提出的研究将（1）评估KO和WT小鼠的酒精消耗和偏好，（2）确定急性补充OC对KO小鼠中ETOH饮用的影响，（3）评估ETOH饮酒。对约束应激的荷尔蒙反应，以及（4）通过在包括交感神经节，脊髓和大脑在内的骨架外组织中免疫定位来探测OC的探针。 意义：迄今为止：POC主要用作骨形成的生物标志物，但是通过EtOH摄入和各种心理压力源以无法归因于成骨细胞活性的方式会持续改变。关于EtoH摄入和压力关联的文献引起了争议。预计拟议的实验将为这一有争议的领域提供至少一个潜在的答案。响应严重的应激源响应ETOH摄入的激素变化可能在KO小鼠中有所不同。因此，我们期望为应激摄入的应力变化提供新的机制提供初步证据。 OC相互作用与各种神经位置的免疫细胞化学通过免疫细胞化学的阳性鉴定将提供有关其在调节酒精摄入的潜在作用部位的基本信息。此外，提出的实验将证明POC不是等离子体的被动组成部分，而是从骨骼发出的反馈回路的一部分，以调节涉及EtOH消耗和对压力条件的反应的神经输入。