Detection of a novel intraneuronal pool of insoluble amyloid β protein that accumulates with time in culture

被引:261
作者
Skovronsky, DM [1 ]
Doms, RW [1 ]
Lee, VMY [1 ]
机构
[1] Univ Penn, Sch Med, Dept Pathol & Lab Med, Ctr Neurodegenerat Dis Res, Philadelphia, PA 19104 USA
关键词
D O I
10.1083/jcb.141.4.1031
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The amyloid-beta peptide (A beta) is produced at several sites within cultured human NT2N neurons with A beta 1-42 specifically generated in the endoplasmic reticulum/intermediate compartment. Since A beta is found as insoluble deposits in senile plaques of the AD brain, and the A beta peptide can polymerize into insoluble fibrils in vitro, we examined the possibility that A beta 1-40, and particularly the more highly amyloidogenic A beta 1-42, accumulate in an insoluble pool within NT2N neurons. Remarkably, we found that formic acid extraction of the NT2N cells solubilized a pool of previously undetectable A beta that accounted for over half of the total intracellular A beta. A beta 1-42 was more abundant than A beta 1-40 in this pool, and most of the insoluble A beta 1-42 was generated in the endoplasmic reticulum/intermediate compartment pathway. High levels of insoluble A beta were also detected in several nonneuronal cell lines engineered to overexpress the amyloid-beta precursor protein. This insoluble intracellular pool of A beta was exceptionally stable, and accumulated in NT2N neurons in a time-dependent manner, increasing 12-fold over a 7-wk period in culture. These novel findings suggest that A beta amyloidogenesis may be initiated within living neurons rather than in the extracellular space. Thus, the data presented here require a reexamination of the prevailing view about the pathogenesis of A beta deposition in the AD brain.
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页码:1031 / 1039
页数:9
相关论文
共 36 条
[31]   Amyloid beta protein in plasma from patients with sporadic Alzheimer's disease [J].
Tamaoka, A ;
Fukushima, T ;
Sawamura, N ;
Ishikawa, K ;
Oguni, E ;
Komatsuzaki, Y ;
Shoji, S .
JOURNAL OF THE NEUROLOGICAL SCIENCES, 1996, 141 (1-2) :65-68
[32]   Amyloids beta(40) and beta(42) are generated intracellularly in cultured human neurons and their secretion increases with maturation [J].
Turner, RS ;
Suzuki, N ;
Chyung, ASC ;
Younkin, SG ;
Lee, VMY .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (15) :8966-8970
[33]   HUMAN NEURONS DERIVED FROM A TERATOCARCINOMA CELL-LINE EXPRESS SOLELY THE 695-AMINO ACID AMYLOID PRECURSOR PROTEIN AND PRODUCE INTRACELLULAR BETA-AMYLOID OR A4 PEPTIDES [J].
WERTKIN, AM ;
TURNER, RS ;
PLEASURE, SJ ;
GOLDE, TE ;
YOUNKIN, SG ;
TROJANOWSKI, JQ ;
LEE, VMY .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (20) :9513-9517
[34]   Intracellular generation and accumulation of amyloid beta-peptide terminating at amino acid 42 [J].
WildBode, C ;
Yamazaki, T ;
Capell, A ;
Leimer, U ;
Steiner, H ;
Ihara, Y ;
Haass, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (26) :16085-16088
[35]   Generation of Alzheimer beta-amyloid protein in the trans-Golgi network in the apparent absence of vesicle formation [J].
Xu, HX ;
Sweeney, D ;
Wang, R ;
Thinakaran, G ;
Lo, ACY ;
Sisodia, SS ;
Greengard, P ;
Gandy, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (08) :3748-3752
[36]   INTRACELLULAR A-BETA-1-42 AGGREGATES STIMULATE THE ACCUMULATION OF STABLE, INSOLUBLE AMYLOIDOGENIC FRAGMENTS OF THE AMYLOID PRECURSOR PROTEIN IN TRANSFECTED CELLS [J].
YANG, AJ ;
KNAUER, M ;
BURDICK, DA ;
GLABE, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (24) :14786-14792