Overproduction of perlecan core protein in cultured cells and transgenic mice.
Heparan sulphate proteoglycan (HSPG) and amyloid P component are the only macromolecules consistently associated with all varieties of amyloid, irrespective of the type of amyloid protein, suggesting that HSPG may play a pathogenetic role in amyloid formation through a common mechanism. In the case of Alzheimer's disease (AD), HSPG, such as perlecan, co-accumulates with amyloid-beta protein (Abeta), a main constituent of amyloid plaques, and paired helical filaments (PHFs). Additionally, in vitro, HSPG accelerates both Abeta fibril and PHF formation and protects Abeta from degradation. Therefore, this study first established lines of P19 mouse embryonic carcinoma cells stably carrying an expression vector encoding the complete perlecan core protein (approximately 400 kD). In the cell lysates, overexpressed perlecan was identified as an approximately 400 kD protein without glycosaminoglycan side-chains, while in the media, secreted perlecan was mostly glycosylated, suggesting that the secretion and glycosylation of perlecan are coupled. Next, transgenic mice were produced using the same expression vector. Marked perlecan overexpression occurred in the cytoplasm of multiple tissues including the brain, heart, kidney, and pancreas, without a discernible increase of perlecan in extracellular matrices. The transgenic mice up to 18 months of age did not develop amyloid or AD-like pathology in the brain or elsewhere, based on histochemical and immunohistochemical analyses. Thus, overproduction of perlecan core protein is insufficient to lead to amyloidosis and AD-like pathology.
Hart
M
M
Department of Comparative Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Li
L
L
Tokunaga
T
T
Lindsey
J R
JR
Hassell
J R
JR
Snow
A D
AD
Fukuchi
K
K
D000544:Alzheimer Disease / Q000378:metabolism*; D000686:Amyloidosis / Q000378:metabolism*; D000818:Animals; D001923:Brain Chemistry; D018236:Carcinoma, Embryonal / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry; D004730:Endothelium, Vascular / Q000737:chemistry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D019812:Heparan Sulfate Proteoglycans / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D007668:Kidney / Q000737:chemistry; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D009206:Myocardium / Q000737:chemistry; D009474:Neurons / Q000737:chemistry; D010179:Pancreas / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000737:chemistry; D014407:Tumor Cells, Cultured / Q000378:metabolism
D019812:Heparan Sulfate Proteoglycans; D012333:RNA, Messenger; C071927:perlecan