Increased Expression of β‐Amyloid Protein Precursor and Microtubule‐Associated Protein τ During the Differentiation of Murine Embryonal Carcinoma Cells

Fukuchi, Ken-ichiro; Deeb, Samir S.; Kamino, Kouzin; Ogburn, Charles E.; Snow, Alan D.; Sekiguchi, Raymond T.; Wight, Thomas N.; Piussan, Henri; Martin, George M.

doi:10.1111/j.1471-4159.1992.tb10063.x

Cited by 40 publications

(21 citation statements)

References 61 publications

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“…3). This finding is in accord with the recent report of Fukuchi et al (43), who have similarly reported an up-regulation of JAPP in P19 cells upon RA-induced differentiation. We extended this analysis to primary hippocampal neurons, demonstrating that 3APP695 expression is also correlated with neuronal differentiation in a different cell culture paradigm (Fig.…”

Section: Discussionsupporting

confidence: 93%

Increased expression of beta-amyloid precursor protein during neuronal differentiation is not accompanied by secretory cleavage.

Hung

Koo

Haass

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

154

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Despite increasing evidence for a pathogenetic role for the fi-amyloid precursor protein KNAPP) in Alzheimer disease, the physiological function of the protein remains unclear. The expression of the neural-specific isoform containing 695 amino acids, BAPP695, is consistent with a role for the protein in neuronal development. In this study, we analyzed the expression of .8APP during the retinoic acidinduced neuronal differentiation of P19 murine embryonal carcinoma cells. Northern blot and RNase protection analyses show a selective increase in I3APP695 expression, concomitant with the morphologic differentiation of P19-derived neurons. Moreover, the time course of increase observed for the .BAPP695 mRNA is paralleled by other neuronal-specific transcripts. A similar increase in (3APP695 is observed at the protein level. Furthermore, we show that levels of 1BAPP695 protein progressively increase during the in vitro differentiation of primary hippocampal neurons. The rmding that fiAPP695 increases selectively and progressively during neuronal differentiation in two different cell culture systems suggests that this isoform has an important cellular function during this process in the brain. Unlike (3APP in most peripheral cell types, the increased levels of IIAPP found in terminally differentiated neuronal cells are not processed in signfcant amounts by secretory cleavage. Thus, differentiation of neurons is accompanied by increased fBAPP695 expression and membrane retention of the protein as intact, full-length molecules that could serve as potential substrates for amyloidogenesis.One of the pathologic hallmarks of Alzheimer disease is the deposition of fibrillar aggregates of a 40-to 43-amino acid polypeptide, the ./A4 peptide, within the brain parenchyma and the walls of the cerebral vasculature (1-3). This -4-kDa protein is derived by proteolytic cleavage from a larger glycosylated membrane-spanning precursor molecule, the p-amyloid precursor protein (J3APP) (4), and is comprised of the 28 amino acids immediately outside the membrane plus the first 12-15 amino acids of the predicted transmembrane domain.Although 8APP transcripts are expressed ubiquitously in all tissues, there is increasing evidence for a unique role for the protein within the nervous system. BAPP mRNA levels are highest within the brain (5-7). In addition, the distribution of one alternatively spliced message encoding a protein of695 amino acids (f3APP695) is largely restricted to neural cells (7,8 (17) and Drosophila APPL suggest that they might be functional homologs.To examine the expression of,APP during neuronal differentiation, we have taken advantage of the P19 mouse embryonal carcinoma cell line (18,19). When these cells are grown under normal conditions, they proliferate as undifferentiated cells. However, in the presence of retinoic acid (RA), they are induced to differentiate into large numbers of neurons and astrocytes as well as a smaller population of fibroblast-like cells (20). Furthermore, the antimetabolite cytosine ara...

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Section: Discussionsupporting

confidence: 93%

Increased expression of beta-amyloid precursor protein during neuronal differentiation is not accompanied by secretory cleavage.

Hung

Koo

Haass

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

154

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“…pGEX-LRPC101 (4445-4544), coding for a fusion protein between GST and the 101 C-terminal residues of human low density lipoprotein receptor-related protein 1 (LRP), was PCR subcloned from pcDNA3.1-mLRP4T100, a gift from Dr. Guojun Bu (Washington University, St Louis). Constructs of pCA-APP695, pCA-S␤C, and pCA-APP⌬AICD were obtained from previous studies (30). All insert sequences were verified by DNA sequencing.…”

Section: Methodsmentioning

confidence: 99%

Endoproteolytic Cleavage of FE65 Converts the Adaptor Protein to a Potent Suppressor of the sAPPα Pathway in Primates

Wang

Yang

et al. 2005

Journal of Biological Chemistry

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Adaptor protein FE65 (APBB1) specifically binds to the intracellular tail of the type I transmembrane protein, ␤-amyloid precursor protein (APP). The formation of this complex may be important for modulation of the processing and function of APP. APP is proteolytically cleaved at multiple sites. The cleavages and their regulation are of central importance in the pathogenesis of dementias of the Alzheimer type. In cell cultures and perhaps in vivo, secretion of the ␣-cleaved APP ectodomain (sAPP␣) is the major pathway in the most cells. Regulation of the process may require extracellular/intracellular cues. Neither extracellular ligands nor intracellular mediators have been identified, however. Here, we show novel evidence that the major isoform of FE65 (97-kDa FE65, p97FE65) can be converted to a 65-kDa N-terminally truncated C-terminal fragment (p65FE65) via endoproteolysis. The cleavage region locates immediately after an acidic residue cluster but before the three major protein-protein binding domains. The cleavage activity is particularly high in human and non-human primate cells and low in rodent cells; the activity appears to be triggered/enhanced by high cell density, presumably via cell-cell/cell-substrate contact cues. As a result, p65FE65 exhibits extraordinarily high affinity for APP (up to 40-fold higher than p97FE65) and potent suppression (up to 90%) of secretion of sAPP␣. Strong p65FE65-APP binding is required for the suppression. The results suggest that p65FE65 may be an intracellular mediator in a signaling cascade regulating ␣-secretion of APP, particularly in primates.FE65 is a multimodular adaptor protein, consisting of three major protein-protein interaction domains, a WW domain and two phosphotyrosine interaction domains (PID) 1 (1, 2) ( Fig. 2A). The interaction between FE65 and APP mainly takes place between the C-terminal PID (PID2) and the APP intracellular domain (AICD), although other regions may also potentially contribute to the action. The physiological effects of FE65-〈PP complexes are not well understood. Genetic evidence suggests that strong FE65-〈PP binding was favored by natural selection during animal evolution toward human lineage (3). 〈PP is a widely studied protein because of its role in the pathogenesis of dementias of the Alzheimer type (DAT). FE65 is predominantly expressed in central nervous system neurons, and its expression is regulated during development and aging, with high levels corresponding to the timing of neural tissue formation and high neuronal activity (4 -8). Selective knockout of the p97FE65 isoform results in modest impairments in learning but severe deficits in relearning spatial tasks (8). These phenotypes reflect reduced synaptic plasticity. Thus, the APP-FE65 pathway may play roles in normal and abnormal cognitive functions. In addition to binding to AICD, FE65 can also interact, at least in vitro, with several other proteins through its WW and PID1 domains (1, 2). FE65 is able to translocate between the nucleus and cytoplasm, consistent with the d...

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“…P1916 mouse embryonic carcinoma cells, and their clonal subline, P19 695B2 cells17, were cultured as described previously16. P19 and P19 695B2 cells were co‐transfected with pCA‐DI‐V and the hygromycin B phosphotransferase gene at a molar ratio of 10:1 using Lipofectamine (Life Technologies, Gaithersburg, MD, USA) according to the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

Overproduction of perlecan core protein in cultured cells and transgenic mice

Hart

Tokunaga

et al. 2001

J. Pathol.

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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.

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Increased Expression of β‐Amyloid Protein Precursor and Microtubule‐Associated Protein τ During the Differentiation of Murine Embryonal Carcinoma Cells

Cited by 40 publications

References 61 publications

Increased expression of beta-amyloid precursor protein during neuronal differentiation is not accompanied by secretory cleavage.

Increased expression of beta-amyloid precursor protein during neuronal differentiation is not accompanied by secretory cleavage.

Endoproteolytic Cleavage of FE65 Converts the Adaptor Protein to a Potent Suppressor of the sAPPα Pathway in Primates

Overproduction of perlecan core protein in cultured cells and transgenic mice

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