ADF/cofilin-actin rods in neurodegenerative diseases

ADF/丝切蛋白-肌动蛋白棒在神经退行性疾病中的作用

基本信息

批准号：
7591023
负责人：
JAMES R BAMBURG
金额：
$ 32.12万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-12-15 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7591023
关键词：
ATP Hydrolysis Actins Affect Affinity Aftercare Alzheimer&apos s Disease American Amyloid Amyloid beta-Protein Amyloid beta-Protein Precursor Animals Apoptosis Binding Brain Brain Diseases Brain region Cell Culture Techniques Cells Cerebellum Cleaved cell Culture Media Deposition Disease Disease Progression Distal Dose Endocytosis Glutamates Grant Growth Hippocampus (Brain)Human Injury Intervention Ischemia Lead Mediating Membrane Lipids Mitochondria Modeling Molecular Conformation Mus Mutation Nature Nerve Growth Factor Receptors Neurites Neurodegenerative Disorders Neurons Neuropil Niemann-Pick Diseases Pathway interactions Peptides Peroxides Pertussis Toxin Process Production Property Proteins Public Health Quality of life Rattus Role Senile Plaques Shapes Side Site Slice Small Interfering RNA Stress Structure Surface Synapses Testing Tg2576 Transgenic Mice Transport Vesicles Vesicle Western Blotting beta secretase cofilin conformer immunoreactivity mutant prevent protein aggregate response retinal rods secretase transcriptional coactivator p75

项目摘要

Within neurites of nearly all cultured hippocampal neurons, transient ATP depletion rapidly inducesrod- shaped structures composed primarily of actin and ADF/cofilin (AC). Rod formation, which sequesters a portion of the actin but virtually all of the AC, is transiently beneficial to the stressed neuron because it spares ATP associated with actin turnover. However, rods can completely occlude the neurite, blocking transport and causing distal neurite withering. Rods are prominent features of Alzeimer's disease (AD) brain but not of control human brain lacking amyloid plaques. Similar structures are found in brains of animals with Niemann-Pick disease typed (NPC1) and of transgenic mice (Tg2576) expressing mutant human amyloid precursor protein (APP). In cultured neurons and mouse brain slices, rods are induced by ischemia, peroxide, NO, and excitotoxic glutamate. In up to 20% of hippocampal neurons, whether from region CA1 or CAS, the AD amyloid beta peptide (Ab) also induces rods: induction is dose-dependent, occuring within 6 h after treatment and plateauing 12-24 h later. As little as 10 nM of Ab oligomer has a significant effect compared to the scrambled peptide control. The nature of the sensitivity of only a subset of neurons to Ab will be explored. Rods block vesicular transport of APP. APP-containing vesicles accumulate at the ends and sides of rods. Within these stalled vesicles is beta-secretase cleaved APP, suggesting that these may be sites of Ab production and/or conversion into more damaging conformers. Taken together, these results suggest a model for AD in which neuronal stress, including Ab formed in familial AD, induces rods that stall vesicle transport and increase toxic Ab, thus inducing rods in neighboring cells. Such a model could explain the formation of amyloid plaques, which would enlarge around the initial site of injury. Using cell culture and organotypic brain slices, we will determine: 1) what activities of cofilin are required for rod formation; 2) if mutations in AC can be identified that prevent rod formation; 3) if rods promote the production or oligomerization of Ab; 4) what makes a subset of neurons sensitive to Ab: and 5) how organotypic brain slices can be used as a model to study where rods form and how they disrupt synapses. Relevance to public health: AD dramatically impacts life quality of senior Americans, affecting 25% of those > 85. This proposal tests a new hypothesis for AD progression and identifies possible sites for targeted intervention.

在几乎所有培养的海马神经元的神经突内，短暂的 ATP 耗竭会迅速诱导杆状细胞主要由肌动蛋白和 ADF/丝切蛋白 (AC) 组成的形状结构。棒状结构，隔离肌动蛋白的一部分，但实际上是全部 AC，对受应激的神经元暂时有益，因为它保留与肌动蛋白周转相关的 ATP。然而，杆可以完全闭塞神经突，阻塞运输并导致远端神经突枯萎。杆状细胞是阿尔茨海默病 (AD) 大脑的显着特征但无法控制缺乏淀粉样斑块的人类大脑。在动物的大脑中也发现了类似的结构患有尼曼-匹克病类型（NPC1）和表达突变人类的转基因小鼠（Tg2576）淀粉样前体蛋白（APP）。在培养的神经元和小鼠脑切片中，杆状细胞是由缺血诱导的，过氧化物、NO 和兴奋毒性谷氨酸盐。在高达 20% 的海马神经元中，无论是来自 CA1 区还是 CAS，AD 淀粉样蛋白 β 肽 (Ab) 也可诱导杆状细胞：诱导呈剂量依赖性，在 6 小时内发生治疗后12-24小时后达到稳定水平。小至 10 nM 的 Ab 寡聚体即可产生显着效果与乱序肽对照相比。仅一部分神经元对抗体敏感的本质将被探索。杆阻断 APP 的囊泡运输。含有APP的囊泡在末端聚集和杆的侧面。在这些停滞的囊泡内是 β-分泌酶裂解的 APP，这表明这些可能是抗体产生和/或转化为更具破坏性的构象异构体的位点。综合起来，这些结果提出了一种 AD 模型，其中神经元应激（包括家族性 AD 中形成的抗体）会诱导视杆细胞停滞囊泡运输并增加有毒抗体，从而在邻近细胞中诱导杆状体。这样的模型可以解释淀粉样斑块的形成，该斑块会在最初受伤部位周围扩大。使用细胞培养和器官型脑切片，我们将确定：1）丝切蛋白的哪些活性是杆形成所必需的； 2）如果可以鉴定出 AC 突变阻止了视杆细胞的形成； 3) 如果棒促进生产或 Ab 寡聚化； 4) 是什么让一部分神经元对抗体敏感：以及 5) 器官型大脑如何切片可以用作模型来研究视杆细胞的形成位置以及它们如何破坏突触。相关性公共卫生：AD 极大地影响了美国老年人的生活质量，影响了 85 岁以上老年人中的 25%。该提案测试了 AD 进展的新假设，并确定了有针对性的干预的可能部位。

项目成果

期刊论文数量（38）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Growth cone-like waves transport actin and promote axonogenesis and neurite branching.

DOI：
10.1002/dneu.20734
发表时间：
2009-10
期刊：
DEVELOPMENTAL NEUROBIOLOGY
影响因子：
3
作者：
Flynn, Kevin C.;Pak, Chi W.;Shaw, Alisa E.;Bradke, Frank;Bamburg, James R.
通讯作者：
Bamburg, James R.

Production and use of replication-deficient adenovirus for transgene expression in neurons.

复制缺陷型腺病毒的生产和使用用于神经元转基因表达。

DOI：
10.1016/s0091-679x(03)01019-7
发表时间：
2003
期刊：
Methods in cell biology
影响因子：
0
作者：
Minamide,LS;Shaw,AE;Sarmiere,PD;Wiggan,O;Maloney,MT;Bernstein,BarbaraW;Sneider,JM;Gonzalez,JA;Bamburg,JamesR
通讯作者：
Bamburg,JamesR

Introduction to cytoskeletal dynamics and pathfinding of neuronal growth cones.

细胞骨架动力学简介和神经元生长锥寻路。

DOI：
10.1177/002215540305100401
发表时间：
2003
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
0
作者：
Bamburg,JamesR
通讯作者：
Bamburg,JamesR

Incorporation of cofilin into rods depends on disulfide intermolecular bonds: implications for actin regulation and neurodegenerative disease.

DOI：
10.1523/jneurosci.6020-11.2012
发表时间：
2012-05-09
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Bernstein BW;Shaw AE;Minamide LS;Pak CW;Bamburg JR
通讯作者：
Bamburg JR

Mapping cofilin-actin rods in stressed hippocampal slices and the role of cdc42 in amyloid-beta-induced rods.