Introduction [4,16], which are considered to play an important role in
Brain Research 888 2001 256–262 www.elsevier.com locate bres
Research report
Immunohistochemical distribution of the receptor for advanced glycation end products in neurons and astrocytes in Alzheimer’s
disease
a , a
a a
a
Nobuyuki Sasaki , Sadamu Toki , Hiroshi Chowei , Toshikazu Saito , Norihito Nakano ,
a b
c
Yorihide Hayashi , Masayoshi Takeuchi , Zenji Makita
a
Department of Neuropsychiatry , Sapporo Medical University, South 1, West 16, Chuo-ku, Sapporo, 060-8543 Japan
b
Department of Biochemistry , Hokuriku University, Kanazawa, Japan
c
Fourth Department of Internal Medicine , Kurume University, Kurume, Japan
Accepted 3 October 2000
Abstract
Advanced glycation end products AGE and the receptor for AGE RAGE have been implicated in the chronic complications of diabetes mellitus DM, and have been reported to play an important role in the pathogenesis of Alzheimer’s disease AD. In this study,
we established a polyclonal anti-RAGE antibody, and examined the immunohistochemical localization of amyloid b protein Ab, AGE, and RAGE in neurons and astrocytes from patients with AD and DM. Our anti-RAGE antibody recognized full-length RAGE 50 kd and
N-terminal RAGE 35 kd in human brain tissue. Ab-, AGE-, and RAGE-positive granules were identified in the perikaryon of hippocampal neurons especially from CA3 and CA4 in all subjects. The distribution and staining pattern of these immunopositive
granules showed good concordance with each antibody. In AD, most astrocytes contained both AGE-and RAGE-positive granules and their distribution was almost the same. Ab-positive granules were less common, but Ab-, AGE-, and RAGE-positive granules were
colocalized in one part of a single astrocyte. In DM patients and control cases, AGE-and RAGE-positive astrocytes were very rare. These finding support the hypothesis that glycated Ab is taken up via RAGE and is degraded through the lysosomal pathway in astrocytes. In
addition to the presence of AGE, the process of AGE degradation and receptor-mediated reactions may contribute to neuronal dysfunction and promote the progression of AD.
2001 Elsevier Science B.V. All rights reserved.
Theme : Disorders of the nervous system
Topic : Degenerative disease: Alzheimer’s disease – beta amyloid
Keywords : Alzheimer’s disease; Advanced glycation end product; Receptor for advanced glycation end product; Astrocyte; Amyloid beta protein
1. Introduction [4,16], which are considered to play an important role in
the pathogenesis of the chronic complications of diabetes Glucose and other reducing sugars react nonenzymati-
mellitus DM [1,2]. After several components of receptor cally with protein amino groups to initiate a posttransla-
for AGE RAGE were characterized [17], it was also tional
modification process
known as
nonenzymatic shown that RAGE and AGE play an important role in
glycosylation [4,10,16]. This reaction proceeds from re- diabetic nephropathy and atherosclerotic vasculopathy
versible Schiff bases to stable, covalently bonded Amadori [13,18,25].
rearrangement products [4]. Once formed, the Amadori Alzheimer’s disease AD is the most common cause of
products undergo further chemical rearrangement to form dementia in Western countries and in Japan. Pathological-
irreversibly bound advanced glycation end products AGE ly, AD is characterized by the presence of senile plaques
SPs, neurofibrillary tangles NFTs, and sever gliosis. AGE can be identified immunohistochemically in both SPs
Corresponding author. Tel.: 181-11-611-2111 ext 3518; fax: 181-
and NFTs [20,21]. Furthermore, RAGE is expressed by
11-644-3041. E-mail address
: nsasakisapmed.ac.jp N. Sasaki.
neurons, microglial cells, and astrocytes in the normal
0006-8993 01 – see front matter
2001 Elsevier Science B.V. All rights reserved. P I I : S 0 0 0 6 - 8 9 9 3 0 0 0 3 0 7 5 - 4
N . Sasaki et al. Brain Research 888 2001 256 –262
257
human brain [3,11], while its expression by cortical glucose-derived AGE collagen [14,15]. This antibody does
neurons increases and becomes more widespread in AD not recognize unmodified RNase, albumin, hemoglobin,
[27]. Since it was reported that RAGE may be the nerve LDL, acetyl LDL, or collagen, as well as previously
cell receptor for amyloid b protein Ab, the role of RAGE reported AGE structures such as 2-furoyl-4 [5]-[2-furanyl]-
in the pathogenesis of AD has attracted considerable 1-H-imidazole
FFI, 1-alkyl-2-formyl-3,4-diglycosyl-
attention [6,11,28]. pyrroles AFGP, pyrraline, pentosidine, or CML [14,15].
We have previous studied the distribution of AGE in AD An anti-Ab antibody 1–42, C-terminal was purchased
and several neurodegenerative diseases, and have sug- from Bachem Feinchemikaline AG Swiss, and a mouse
gested that AGE may be an important factor in the anti-glial fibrillary acidi protein GFAP antibody was
progression of various neurodegenerative disorders [20]. purchased from Dakopatts Denmark.
However, little is known about the detailed role of RAGE A polyclonal antibody against RAGE were raised in
in AD [28]. rabbits. The peptides were synthesized according to the
To investigate the role of RAGE in AD and DM, we amino acid sequences of RAGE, residues 167–180 [6].
established an anti-RAGE antibody and performed im- Synthesized peptide was coupled to keyhole limpet
munohistochemical studies. Our data demonstrated that hemocyanin, and mixed with an equal volume of Freud’s
RAGE was present in astrocytes from AD brains along complete adjuvant. The conjugated peptide were injected
with AGE and Ab, suggesting that RAGE-mediated degra- into rabbits 33 at 2-week intervals. Serum was obtained 2
dation of Ab occurs in astrocytes. weeks after last injection, and antibody titer was assessed
by ELISA. When antibody levels plateaued, the serum was collected and stored at 2808C until required.