Differential expression of fibroblast growth factor-2 and receptor by glial cells in experimental autoimmune encephalomyelitis (EAE)

To assess the expression pattern of basic fibroblast growth factor (FGF-2) and one of its receptors (FGFR-1/flg) during autoimmune inflammation of the CNS, FGF-2, and FGFR1/flg peptide and mRNA levels were examined by immunocytochemistry, by in situ hybridisation and by Northern blot analysis in T cell-mediated EAE of the Lewis rat. In naive control animals as well as in animals injected with nonencephalitogenic, PPD-reactive T lymphocytes, FGF-2 immunoreactivity was low and confined to blood vessels and to a few spinal cord neurons. In rats injected with encephalitogenic, MBP-reactive T lymphocytes, however, FGF-2-immunoreactive cells were detected from day 4 after T cell transfer onward, i.e., from the onset of clinical symptoms. The number of FGF-2 immunoreactive cells was highest between days 6 and 10 after T cell transfer. Increased FGF-2 peptide expression was paralleled by increased FGF-2 mRNA expression on macrophages/microglia in the spinal cord. By 21 days after T cell transfer, i.e. after complete recovery, FGF-2 peptide and mRNA expression had fully subsided. Based on morphological criteria and on double labeling with the macrophage/microglia-binding lectin GSI-B₄ two cell types expressed FGF-2: 1) round macrophages within the core, and 2) activated microglia at the edges of white and grey matter perivascular lesions. Paralleling the temporal and spatial expression pattern of FGF-2, FGFR-1/flg immunoreactivity was induced on activated macrophages/microglia but also on reactive astrocytes bordering perivascular inflammatory lesions. In situ hybridisation analysis furthermore showed that macrophages/microglia expressed the FGFR-1/flg mRNA, and that receptor mRNA expression paralleled ligand mRNA expression. Macrophage/microglia-derived FGF-2 could serve two main functions in EAE: 1) regulate microglial activation in an autocrine fashion, and 2) help to target astrocyte-derived insulin-like growth factor-I (IGF-I) to potentially injured oligodendrocytes in demyelination. © 1996 Wiley-Liss, Inc.     

Cytoarchitecture of fibroblast growth factor receptor 2 (FGFR-2) immunoreactivity in astrocytes of neurogenic and non-neurogenic regions of the young adult and aged rat brain

Fibroblast growth factors (FGFs) are polypeptides that exert diverse biological effects on many cell types and tissues during embryogenesis and adulthood. In the adult brain, FGF-2 is primarily expressed by astrocytes and select groups of neurons. It has been shown that FGF-2 is neuroprotective and can stimulate proliferation of NSCs in neurogenic regions of the adult mammalian brain. Cellular responses to FGFs are mediated through membrane-spanning tyrosine kinase receptors in conjunction with low affinity binding to heparin sulfate proteoglycans. Four FGF receptors (FGFR1-4) have been cloned and characterized to date. In this study, we describe the anatomical distribution of FGFR-2 in young and aged rat brains. We demonstrate that the olfactory bulb, hippocampus, and cerebellum display the most robust FGFR-2 expression and observed age-related decrease in FGFR-2 levels in some but not all brain regions. In addition, we identified astrocytes as the primary source of FGFR-2 expression using immunofluorescence confocal microscopy. The astrocyte populations in the neurogenic areas, the subventricular zone (SVZ) and the subgranular zone (SGZ) of the dentate gyrus, express high levels of FGFR-2 protein, which points to its possible involvement in neurogenesis. We also explored the role of FGFR-2 in response to perforant pathway lesion and observed enhanced FGFR-2 expression by astrocytes surrounding the lesion. Thus, FGF-2 biological effects on astrocytes appear to be mediated through FGFR-2-dependent mechanisms, and this may provide an indirect route by which FGF-2 acts on neuronal populations.     

Increased expression and nuclear accumulation of basic fibroblast growth factor in primary cultured astrocytes following ischemic-like insults.

Basic fibroblast growth factor (bFGF) is a biological active polypeptide with potent trophic effects on neurons, glia and endothelial cells. In the present study, we examined the temporal expression profile of bFGF protein in cultured cortical astrocytes under ischemic-like conditions such as serum-free, glucose-free or glutamate application. A peak increase of bFGF level was observed at 24 h after the initiation of insults. A striking increase in the bFGF immunoreactivity and a moderate increase in the fibroblast growth factor receptor-1 (FGFR-1) immunoreactivity were also found in the astrocytes treated with serum- or glucose-deprivation or glutamate. The increased bFGF immunoreactivity and FGFR-1 immunoreactivity were mainly accumulated in the nuclei of astrocytes. The results suggest that the expression of bFGF and FGFR-1 in the astrocytes, especially in the nuclear interior, can be up-regulated under ischemic-like conditions and that the up-regulation of bFGF and FGFR-1 may play an important role in the maintenance and repair of the central nervous system (CNS) after ischemia.     

nNOS expression in reactive astrocytes correlates with increased cell death related DNA damage in the hippocampus and entorhinal cortex in Alzheimer's disease

The immunocytochemical distribution of the neuronal form of nitric oxide synthase (nNOS) was compared with neuropathological changes and with cell death related DNA damage (as revealed by in situ end labeling, ISEL) in the hippocampal formation and entorhinal cortex of 12 age-matched control subjects and 12 Alzheimer's disease (AD) patients. Unlike controls, numerous nNOS-positive reactive astrocytes were found in AD patients around beta-amyloid plaques in CA1 and subiculum and at the places of clear and overt neuron loss, particularly in the entorhinal cortex layer II and CA4. This is the first evidence of nNOS-like immunoreactivity in reactive astrocytes in AD. In contrast to controls, in all but one AD subject, large numbers of ISEL-positive neuronal nuclei and microglial cells were found in the CA1 and CA4 regions and subiculum. Semiquantitative analysis showed that neuronal DNA fragmentation in AD match with the distribution of nNOS-expressing reactive astroglial cells in CA1 (r = 0.74, P < 0.01) and CA4 (r = 0.58, P < 0.05). A portion of the nNOS-positive CA2/CA3 pyramidal neurons was found to be spared even in the most affected hippocampi. A significant inverse correlation between nNOS expression and immunoreactivity to abnormally phosphorylated tau proteins (as revealed by AT8 monoclonal antibody) in perikarya of these CA2/3 neurons (r = -0.85, P < 0.01) suggests that nNOS expression may provide selective resistance to neuronal degeneration in AD. In conclusion, our results imply that an upregulated production of NO by reactive astrocytes may play a key role in the pathogenesis of AD.     

Fibroblast growth factor-9 modulates the expression of myelin related proteins and multiple fibroblast growth factor receptors in developing oligodendrocytes

The effect of fibroblast growth factor (FGF)-9 on the expression of FGF receptors (FGFR) and the major myelin proteins was examined in cultures of developing rat brain oligodendrocytes (OLs), using immunological techniques. FGFR-1, -3, and -4 were expressed at all developmental stages but were not present in isolated myelin fractions. By contrast, FGFR-2 protein was predominantly localized to differentiating cells and myelin. FGF-9 altered FGFR and myelin protein levels during OL differentiation; there was increased expression of FGFR-1 and decreased levels of both FGFR-2 and myelin proteins. Further, FGF-9 stimulated mitogen-associated protein kinase (MAPK) phosphorylation. The effect of FGF-9 on MAPK, however, was transient and less robust in progenitor cells than in differentiated oligodendrocytes. The effects of FGF-9 and FGF-2 on FGFR and myelin protein levels were comparable; both up-regulated FGFR-1, and down-regulated FGFR-2, CNP, PLP and MBP. These findings suggest that FGF-9 may be important for glial cell development.     

Immunohistochemical analyses of fibroblast growth factor receptor-1 in the human substantia nigra. Comparison between normal and Parkinson's disease cases

The use of neurotrophic growth factors as a means of preventing loss of the dopaminergic (DA) neurons in the substantia nigra (SN) is becoming an accepted treatment strategy for Parkinson's disease (PD). In earlier studies, we showed that there was a selective loss of basic fibroblast growth factor (bFGF) immunoreactivity in DA neurons of the SN in PD suggesting that a deficiency of bFGF might contribute to cell death. As a preliminary step to assessing the potential for using bFGF or its analogs as therapeutic agents, the expression of fibroblast growth factor receptor-1 (FGFR-1) in the SN of normal and PD cases was investigated immunohistochemically. FGFR-1 immunoreactivity could be detected in DA neurons of the SN in young and old neurologically normal cases with an apparent decline with age. FGFR-1 immunoreactivity was also detected in many of the residual SN neurons in most of the idiopathic PD cases. These results indicate that FGFR-1 immunoreactivity, and possibly FGF binding activity, is retained in DA neurons in PD.     

Crotonaldehyde accumulates in glial cells of Alzheimer’s disease brain

Several studies have documented the involvement of oxidative stress represented by lipid peroxidation in the pathogenesis of Alzheimer’s disease (AD). To test whether the highly reactive carbonyl crotonaldehyde (CRA), generated during lipid peroxidation, is involved in AD, we performed an immunohistochemical analysis in AD and age-matched control hippocampi using a specific antibody against protein-bound CRA (P-CRA). In the AD cases, P-CRA immunoreactivity was preferentially localized in reactive astrocytes and microglia around senile plaques (SPs) and those present in the neuropil, while it was weakly detectable in neurons and neurofibrillary tangles. P-CRA immunoreactivity was also localized in all portions of diffuse SPs and the dystrophic neurites of neuritic and classical SPs, but was undetectable in amyloid cores. Age-matched controls showed P-CRA immunoreactivity only very weakly in neurons. In contrast to P-CRA, immunoreactivities for protein-bound acrolein and 4-hydroxy-2-nonenal were mainly localized to neurons and rarely seen in glial cells. Our results suggest that increased oxidative stress and CRA formation in glial cells is implicated in the disease processes of AD.     

FGF-18 is a neuron-derived glial cell growth factor expressed in the rat brain during early postnatal development

We examined the expression of fibroblast growth factor-18 (FGF-18) in the rat brain during postnatal development by in situ hybridization. FGF-18 was transiently expressed at the early postnatal stages in various regions of the rat brain including the cerebral cortex and hippocampus. FGF-18 in the brain was preferentially expressed in neurons but not in glial cells. To elucidate the role of FGF-18 in the brain, we examined the ligand-specificity of FGF-18 by the BIAcore system. FGF-18 was found to bind to FGF receptors (FGFRs)-3c and -2c but not to FGFR-1c, suggesting that FGF-18 acts on glial cells but not on neurons. Therefore, we examined the mitogenic activity of FGF-18 for cultured rat astrocytes and microglia. FGF-18 was found to have mitogenic activity for both astrocytes and microglia. We also examined the neurotrophic activity of FGF-18 for cultured rat cortical neurons. FGF-18 was found to have no neurotrophic activity. The present findings indicated that FGF-18 is a unique FGF that plays a role as a neuron-derived glial cell growth factor in early postnatal development when gliogenesis occurs.     

Immunoreactivity of Presenilin-1 and Tau in Alzheimer''s Disease Brain

Mutations in the presenilin-1 gene (PS-1) on chromosome 14 are causative for early-onset familial Alzheimer's disease (AD). In order to study the localization of PS-1 in human brain, a polyclonal antibody, SB63, against a N-terminal epitope of PS-1 (²⁵VRSQNDNRERQEHND⁴⁰), was raised in rabbits and characterized. Immunolabeling with SB63 of formalin-fixed sections of hippocampus from cases of PS-1-linked AD (PS-1 I143T (AD/A), G384A (AD/B)), sporadic AD, and controls showed a predominant neuronal staining pattern with a stronger immunoreactivity in pyramidal neurons. Staining was mainly granular and localized in the neuronal cell body as well as in neuronal processes. In AD some dystrophic neurites surrounding the amyloid plaques were stained, but no immunoreactivity was observed in the amyloid core. Although PS-1 was present in tangle bearing neurons, colocalization of PS-1 and tau could not be detected using immunofluorescence double labeling. Our data indicate that the pattern of PS-1 immunoreactivity in the hippocampus does not substantially differ between PS-1-linked AD, sporadic AD, and controls.     

Accumulation of a repulsive axonal guidance molecule RGMa in amyloid plaques: a possible hallmark of regenerative failure in Alzheimer's disease brains

J. Satoh, H. Tabunoki, T. Ishida, Y. Saito and K. Arima (2013) Neuropathology and Applied Neurobiology 39, 109–120 Accumulation of a repulsive axonal guidance molecule RGMa in amyloid plaques: a possible hallmark of regenerative failure in Alzheimer's disease brains Aims: RGMa is a repulsive guidance molecule that induces the collapse of axonal growth cones by interacting with the receptor neogenin in the central nervous system during development. It remains unknown whether RGMa plays a role in the neurodegenerative process of Alzheimer's disease (AD). We hypothesize that RGMa, if it is concentrated on amyloid plaques, might contribute to a regenerative failure of degenerating axons in AD brains. Methods: By immunohistochemistry, we studied RGMa and neogenin (NEO1) expression in the frontal cortex and the hippocampus of 6 AD and 12 control cases. The levels of RGMa expression were determined by qRT‐PCR and Western blot in cultured human astrocytes following exposure to cytokines and amyloid beta (Aβ) peptides. Results: In AD brains, an intense RGMa immunoreactivity was identified on amyloid plaques and in the glial scar. In the control brains, the glial scar and vascular foot processes of astrocytes expressed RGMa immunoreactivity, while oligodendrocytes and microglia were negative for RGMa. In AD brains, a small subset of amyloid plaques expressed a weak NEO1 immunoreactivity, while some reactive astrocytes in both AD and control brains showed an intense NEO1 immunoreactivity. In human astrocytes, transforming growth factor beta‐1 (TGFβ₁), Aβ_1–40 or Aβ_1–42 markedly elevated the levels of RGMa, and TGFβ₁ also increased its own levels. Coimmunoprecipitation analysis validated the molecular interaction between RGMa and the C‐terminal fragment β of amyloid beta precursor protein (APP). Furthermore, recombinant RGMa protein interacted with amyloid plaques in situ. Conclusions: RGMa, produced by TGFβ‐activated astrocytes and accumulated in amyloid plaques and the glial scar, could contribute to the regenerative failure of degenerating axons in AD brains.     

Accumulation of protein-bound 4-hydroxy-2-hexenal in spinal cords from patients with sporadic amyotrophic lateral sclerosis

4-Hydroxy-2-hexenal (HHE) is a toxic, reactive aldehydic intermediate formed by nonenzymatic peroxidation of n-3 polyunsaturated fatty acids. The aim of this study was to determine the implication for HHE in the pathomechanism of amyotrophic lateral sclerosis (ALS) by immunohistochemical and enzyme-linked immunosorbent assay (ELISA) techniques using a mouse monoclonal IgG(1) antibody mAbHHE53 specific for protein-bound HHE. Immunohistochemical analysis on formalin-fixed, paraffin-embedded sections and frozen sections of spinal cords obtained at autopsy from 10 sporadic ALS patients and 10 age-matched control subjects demonstrated that protein-bound HHE immunoreactivity was seen and was prominent in the entire gray matter in the ALS cases and localized in the neurons, reactive astrocytes, microglial cells, and the surrounding neuropil, while the immunoreactivity was obscure or undetectable in the control cases. No significant protein-bound HHE immunoreactivity was seen in sections processed with omission of mAbHHE53 or in sections incubated with the antibody with an excess of the respective antigen. Competitive ELISA analysis on trypsin-digested protein extracts of fresh-frozen spinal cord samples disclosed a significant increase in protein-bound HHE level in the ALS cases compared with the control cases. Our results indicate that enhanced HHE formation occurs in the entire gray matter of sporadic ALS spinal cords and suggest that the selective vulnerability of motor neurons to HHE mediates the pathomechanism of this disease.     

Immunohistochemical distribution of the receptor for advanced glycation end products in neurons and astrocytes in Alzheimer's disease

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 beta protein (Abeta), 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. Abeta-, 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. Abeta-positive granules were less common, but Abeta-, 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 Abeta 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.     

Glial expression of fibroblast growth factor-9 in rat central nervous system.

We examined the expression of fibroblast growth factor (FGF)-9 in the rat central nervous system (CNS) by immunohistochemistry and in situ hybridization studies. FGF-9 immunoreactivity was conspicuous in motor neurons of the spinal cord, Purkinje cells, and neurons in the hippocampus and cerebral cortex. In addition to the neuronal localization of FGF-9 immunoreactivity that we reported previously, the present double-label immunohistochemistry clearly demonstrated that the immunoreactivity was present in glial fibrillary acidic protein (GFAP)-positive astrocytes preferentially present in the white matter of spinal cord and brainstem of adult rats and in CNPase-positive oligodendrocytes that were arranged between the fasciculi of nerve fibers in cerebellar white matter and corpus callosum of both adult and young rats. There was a tendency for FGF-9 immunoreactivity in oligodendrocytes to be more pronounced in young rats than in adult rats. The variation of oligodendrocyte FGF-9 immunoreactivity in adult rats was also more pronounced than that in young rats. With in situ hybridization, FGF-9 mRNA was observed in astrocytes in the white matter of rat spinal cord and oligodendrocytes in the white matter of cerebellum and corpus callosum of adult and young rats. The expression of FGF-9 mRNA in glial cells was lower than in neurons, and not all glial cells expressed FGF-9. In the present study, we demonstrated that FGF-9 was expressed not only in neurons but also in glial cells in the CNS. FGF-9 was considered to have important functions in adult and developing CNS.     

bFGF and FGFR-3 immunoreactivity in the rat brain following systemic kainic acid administration at convulsant doses: localization of bFGF and FGFR-3 in reactive astrocytes, and FGFR-3 in reactive microglia

Strong bFGF immunoreactivity was observed in reactive astrocytes, as shown by double-labeling immunohistochemistry of bFGF and GFAP, from days 7 up to 30 (last time point examined) following kainic acid (KA) injection at convulsant doses in the adult rat. bFGF was not found in OX-42-positive reactive microglia. A few reactive glia co-localized FGFR-3 and GFAP, whereas the majority of cells expressing FGFR-3 were OX-42-immunoreactive. This was further supported by the observation that only ≈ 10% of reactive glia co-localized bFGF and FGFR-3. These results show that reactive astrocytes are a major source of bFGF during the subacute stages of tissue damage following KA injection and that reactive astrocytes and, most particularly, reactive microglia are putative targets of bFGF through FGFR-3.     

Estrogen receptor alpha-immunoreactive astrocytes are increased in the hippocampus in Alzheimer's disease

Postmenopausal estrogen use may decrease the risk, and delay the onset and progression, of Alzheimer's disease (AD). By means of fluorescence immunocytochemistry, the present study investigated the distribution of estrogen receptor alpha (ERalpha) in the human hippocampus in controls and in AD cases. ERalpha immunoreactivity was observed in neurons and glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes in the hippocampus both in controls and AD cases. The number and density of GFAP- and ERalpha-positive astrocytes was increased in AD. The number of GFAP-immunoreactive astrocytes, the number of nuclear ERalpha-staining astrocytes, and cytoplasmic ERalpha-staining astrocytes per unit area (1 mm(2)) significantly increased (P < 0.001, P < 0.05, P < 0.05, respectively) in CA1 in AD patients, while the percentage of ERalpha-immunoreactive astrocytes of the two groups did not differ (P > 0.05). These data suggest an important role for ERalpha-mediated effects of estrogens on neurons and astrocytes in the hippocampus of human and AD patients.     

Aquaporin expression in the brains of patients with or without cerebral amyloid angiopathy

Aquaporins have recently been identified as protein channels involved in water transport. These channels may play a role in the edema formation and alterations in microvascular function observed in Alzheimer disease (AD) and cerebral amyloid angiopathy (CAA). We investigated the expression of aquaporin 1 (AQP1) and aquaporin 4 (AQP4) in 24 human autopsy brains consisting of 18 with AD and varying degrees of CAA and 6 with no pathologic abnormalities using immunohistochemistry. In cases of AD and CAA, there was enhanced AQP4 expression compared with the age- and sex-matched controls. Aquaporin 4 immunoreactivity was prominent at the cerebrospinal fluid and brain interfaces, including subpial, subependymal, pericapillary, and periarteriolar spaces. Aquaporin 1 expression in AD and CAA cases was not different from that in age- and sex-matched controls. Double labeling studies demonstrated that both AQP1 and 4 were localized to astrocytes. Both enhanced AQP4 expression and its unique staining pattern suggest that these proteins may be important in the impaired water transport observed in AD and CAA.     

In vivo and in vitro localization of brain-derived neurotrophic factor,fibroblast growth factor-2 and their receptors in the bullfrog vestibular end organs

The inner ear sensory epithelia of vertebrates are composed mainly of supporting cells and hair cells (HCs). Brain-derived neurotrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) are trophins that are believed to play an essential role in the development and innervation of inner ear epithelia. Both trophins also may play a crucial role in the maintenance and regeneration of hair cells in the adult vertebrate ear. In the bullfrog vestibular system, hair cells are produced throughout life, and the epithelia regenerates following ototoxicity. The expression of BDNF and FGF-2 in the vestibular organs of the adult bullfrog was investigated at a cellular level both in histological sections and in vitro in dissociated cell cultures. In histological sections of the crista ampullaris, in situ hybridization and immunocytochemical techniques demonstrated that HCs express both BDNF and its receptor trkB, while the supporting cells express the receptor trkB alone. Following dissociation and in vitro cell culture no changes in the pattern of BDNF and trkB receptor were observed. Immunocytochemical studies demonstrated that in vivo hair cells express FGF-2 and the receptors FGFR-1 and FGFR-2 while supporting cells do not express either molecule. Following dissociation, HCs continue to express FGF-2 and its two receptors, while supporting cells upregulate the expression of FGF-2 and its receptor FGFR-2. These data confirm the potential role of BDNF and FGF-2 trophic regulation of the sensory epithelia of the adult inner ear. The findings suggest that BDNF has a role in the maintenance of the vestibular epithelia while FGF-2 may regulate the proliferation of supporting cells.     

Lamina-selective A68 immunoreactivity in primary visual cortex of Alzheimer's disease patients

Sections of area 17 from 9 Alzheimer's disease (AD) and 13 control cases were stained using the monoclonal antibody, Alz-50, which identifies pathologically-involved neuronal systems in AD patients. Strong positive staining was seen only in AD cases. The pattern of staining was highly lamina-selective; laminae I-IVa and V were intensely stained and lamina VI staining was moderately intense, while laminae IVb and IVc were relatively unstained. The results support the growing consensus that cortical pathology in AD is not generalized, but affects specific neuronal populations. The specific anatomical distribution of degenerative change suggests that there may be a specific set of visual deficits in AD patients.     

Alz-50 immunoreactive neuropil differentiates hippocampal complex subfields in Alzheimer's disease

The topographic distribution of Alz-50 containing profiles was determined within the hippocampal formation and anterior parahippocampal gyrus by using a monoclonal antibody directed against the A68 protein in normal and Alzheimer's diseased (AD) brains. Although there was a paucity of immunoreactive neuropil in the normal hippocampal complex, there were a few Alz-50 positive neurons that occupied the hippocampal subfield, CA2. In most AD cases, Alz-50 immunoreactive neuropil was prominent in the outer two-thirds of the molecular layer of the dentate gyrus, although a few cases exhibited staining in the inner third of the molecular layer. CA2 was characterized by an increased density of neuropil staining within stratum pyramidale. The neuropil in subfield CA1 was stained densely with Alz-50 in strata oriens, pyramidale, and at the border between strata lacunosum-moleculare and radiatum. Alz-50 immunostained neurites occupied primarily the lateral two-thirds of the subiculum proper, whereas only sparse staining was seen in the adjacent presubiculum. Alz-50 neuropil and neuronal staining displayed three distinct laminar patterns along the mediolateral extent of the entorhinal cortex, whereas the perirhinal cortex exhibited a bilaminar pattern of immunoreactivity involving heavy staining in layers 1-3 as compared to layer 5. In general, the density of Alz-50 neurite staining in the neuropil appeared inversely proportional to the distribution of Alz-50 immunoreactivity within dendritic and somal compartments. Interestingly, the patterns of Alz-50 staining observed in the hippocampal complex in AD coincides with patterns of well-characterized afferent fiber pathways to these regions, thus further supporting the suggestion that hippocampal subfield specific pathology effectively disconnects medial temporal structures from adjacent neocortex in AD.     

Immunohistochemical distribution of the receptor for advanced glycation end products in neurons and astrocytes in Alzheimer’s disease

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 β protein (Aβ), 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. Aβ-, 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. Aβ-positive granules were less common, but Aβ-, 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 Aβ 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.