权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Imbalances in Calcium Homeostasis: Mechanism Behind Ethanol-Related Dendritic Deg

钙稳态失衡：乙醇相关树突状脱位背后的机制

基本信息

批准号：
7918174
负责人：
CYNTHIA A DLUGOS
金额：
$ 17.11万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-20 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7918174
关键词：
ATP phosphohydrolase Accidents Acute Address Adult Aftercare Age Aging Alcohol consumption Alcohols Apoptotic Ca(2+)-Transporting ATPase Calcium Calcium Signaling Calcium ion Calcium-Binding Proteins Cell Culture Techniques Cerebellar cortex structure Cerebellum Chronic Cognitive Confocal Microscopy Coupled Dendrites Distal Dose Effectiveness Elderly Ethanol Fluorescence Fura-2 Future GRP78 gene Goals Health Homeostasis Impaired cognition Laboratories Measurement Measures Modeling Motor Neurons Output Pathway interactions Population Preparation Process Proteins Pump Rattus Second Messenger Systems Signal Transduction Slice Smooth Endoplasmic Reticulum Structure System Thapsigargin Time Vertebral column aging population base calreticulin caspase 12 chronic alcohol ingestion density driving force falls feeding frontal lobe functional decline human subject immunocytochemistry in vivo male neurotoxic public health relevance ratiometric response sarcoplasmic reticulum calcium ATPase second messenger smooth endoplasmic reticulum membrane treatment duration

项目摘要

DESCRIPTION (provided by applicant): Consistent, albeit moderate alcohol consumption in human adults results in permanent alterations in the cerebellum that discretely disrupt motor functions and become more pronounced with age. This is a major health concern as the aging population continues to escalate. In aging rats, chronic ethanol consumption results in regression of Purkinje neuron (PN) dendrites, a process that surely alters cerebellar function as PN are the sole output neurons from the cerebellum. Ethanol-related dendritic regression is coupled with other degenerative changes in dendrites such as dilation of the smooth endoplasmic reticulum (SER), the major calcium storage component of PN dendrites. Dilation of the SER in PN dendrites is strongly suggestive of increases in intradendritic calcium, a second messenger signal for a myriad of neuronal pathways. Ethanol-related dilation of the SER also suggests that ethanol-related alterations may have occurred in the sarcoplasmic reticulum calcium ATPase (SERCA) pump that resequesters calcium into the SER lumen following calcium signaling. The overall goal of this proposal is to show that ethanol-related increases in intracellular calcium levels and declines in the levels and/ or function of the SERCA 2b calcium ATPase pump may be the driving force that culminates in PN dendritic degeneration in aging and alcohol-fed rats. Calreticulin, a calcium binding protein that interacts with the SERCA pump will also be examined. To achieve this goal, 12 mo old F344 male rats will be fed the AIN-93M ethanol (EF rats), AIN-93M control (PF) rats, or rat chow (CF rats) for a period of weeks. Young control rats will also be included in the analysis. To measure calcium within dendrites and specific dendritic components, acute cerebellar slices will be made following 10, 20, or 40 weeks of ethanol treatment, electroporated to facilitate Fura-2 entry into PN, and ratiometric calcium measures made on the slices. Levels of the SERCA 2b ATPase pump and calreticulin will be assessed with confocal microscopy, and fluorescence intensity correlation after treatment durations of 10, 20 or 40 weeks in proximal and distal dendritic branches, spines, and branchpoints. SERCA 2b pump function will be assessed by observing the ability of the PN to handle large calcium transients when calcium sequestration systems other than the SERCA pump are blocked and by assessing whether ethanol predisposes the neuron to thapsigargin-induced expression of caspase-12.and GRP78 It is predicted that chronic ethanol consumption will result in increases in intracellular calcium ions that will be related to ethanol-related declines in the function or expression of the SERCA 2b pump. Results from this study will provide a basis for future studies of the actions of ethanol on intracellular calcium homeostasis and the mechanisms that result in PN dendritic degeneration. PUBLIC HEALTH RELEVANCE: The elderly U.S. population will double by 2050, an escalation accompanied by a tremendous increase in the number of elderly lifetime alcohol consumers. Recent studies show that consistent, albeit moderate alcohol consumption in human adults results in permanent alterations in cerebellar- based motor coordination and also in the cognitive activities ascribed to the cerebellum's connections with the frontal lobe. Permanent deficits result from these alterations that become more prevalent with age and predispose our increasing elderly population to falls, accidents, and cognitive dysfunctions.

描述（由申请人提供）：成年人持续适度饮酒导致小脑永久性改变，离散地破坏运动功能，并随着年龄增长而变得更加明显。随着人口老龄化的不断加剧，这是一个重大的健康问题。在老龄大鼠中，慢性乙醇消耗导致浦肯野神经元（PN）树突的退化，这一过程肯定会改变小脑功能，因为PN是小脑唯一的输出神经元。乙醇相关的树突退化与树突中的其他退行性变化相结合，例如滑面内质网（SER）扩张，滑面内质网是PN树突的主要钙储存组分。PN树突中SER的扩张强烈提示树突内钙的增加，这是无数神经元通路的第二信使信号。乙醇相关的SER扩张也表明，乙醇相关的改变可能发生在肌浆网钙ATP酶（SERCA）泵，重新分配钙到SER腔后，钙信号。本提案的总体目标是表明，乙醇相关的细胞内钙水平增加和SERCA 2b钙ATP酶泵的水平和/或功能下降可能是衰老和酒精喂养大鼠PN树突状变性的驱动力。还将检查钙网蛋白，一种与SERCA泵相互作用的钙结合蛋白。为了实现这一目标，12月龄的F344雄性大鼠将喂食AIN-93 M乙醇（EF大鼠）、AIN-93 M对照（PF）大鼠或大鼠饲料（CF大鼠）数周。幼龄对照大鼠也将纳入分析中。为了测量树突和特定树突成分内的钙，将在乙醇处理10、20或40周后制备急性小脑切片，电穿孔以促进Fura-2进入PN，并在切片上进行比率钙测量。在治疗持续时间为10、20或40周后，将使用共聚焦显微镜和荧光强度相关性评估近端和远端树突状分支、棘和分支点中SERCA 2b ATP酶泵和钙网蛋白的水平。SERCA 2b泵的功能将通过观察当除SERCA泵以外的钙螯合系统被阻断时PN处理大的钙瞬变的能力以及通过评估乙醇是否使神经元倾向于毒胡萝卜素诱导的半胱天冬酶-12和GRP 78的表达来评估。SERCA 2b泵的功能或表达的相关下降。本研究结果将为进一步研究乙醇对细胞内钙稳态的作用以及导致PN树突变性的机制提供基础。公共卫生关系：到2050年，美国老年人口将翻一番，伴随着老年人终生饮酒人数的大幅增加。最近的研究表明，成年人持续适度饮酒会导致小脑运动协调性的永久性改变，以及小脑与额叶连接的认知活动。永久性缺陷是由这些改变引起的，这些改变随着年龄的增长而变得更加普遍，并使我们不断增加的老年人口更容易发生福尔斯、事故和认知功能障碍。