Immunohistochemical analysis of presenilin‐1 expression in the mouse brain

Moussaoui, Saliha; Czech, Christian; Pradier, Laurent; Blanchard, Véronique; Bonici, Bruno; Gohin, M.; Impérato, Assunta; Revah, Frédéric

doi:10.1016/0014-5793(96)00250-5

Cited by 56 publications

(29 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These posttranslational modifications are thereby also excluded as contributing to the formation of the high molecular mass, SDS-resistant aggregates (100 -250 kDa) in SDS-PAGE electrophoresis. These aggregates were observed to a variable extent in all our experiments, and were noticed by others using other cell types or brain tissue (36,38,39). Since the smears were detected with three different antibodies recognizing three different epitopes, and by a Myc-directed mAb using a Myctagged PS1 construct, the aggregates must contain presenilin core proteins, alone or associated with unidentified components.…”

Section: Discussionsupporting

confidence: 78%

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Strooper¹,

Beullens²,

Contreras

et al. 1997

Journal of Biological Chemistry

272

187

View full text Add to dashboard Cite

Presenilins 1 and 2 are unglycosylated proteins with apparent molecular mass of 45 and 50 kDa, respectively, in transfected COS-1 and Chinese hamster ovary cells. They colocalize with proteins from the endoplasmic reticulum and the Golgi apparatus in transfected and untransfected cells. In COS-1 cells low amounts of intact endogeneous presenilin 1 migrating at 45 kDa are detected together with relative larger amounts of presenilin 1 fragments migrating between 18 and 30 kDa. The presenilins have a strong tendency to form aggregates (mass of 100 -250 kDa) in SDS-polyacrylamide gel electrophoresis, which can be partially resolved when denatured by SDS at 37°C instead of 95°C. Sulfation, glycosaminoglycan modification, or acylation of the presenilins was not observed, but both proteins are posttranslationally phosphorylated on serine residues. The mutations Ala-246 3 Glu or Cys-410 3 Tyr that cause Alzheimer's disease do not interfere with the biosynthesis or phosphorylation of presenilin 1. Finally, using low concentrations of digitonin to selectively permeabilize the cell membrane but not the endoplasmic reticulum membrane, it is demonstrated that the two major hydrophilic domains of presenilin 1 are oriented to the cytoplasm. The current investigation documents the posttranslational modifications and subcellular localization of the presenilins and indicates that postulated interactions with amyloid precursor protein metabolism should occur in the early compartments of the biosynthetic pathway.

show abstract

Section: Discussionsupporting

confidence: 78%

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Strooper¹,

Beullens²,

Contreras

et al. 1997

Journal of Biological Chemistry

272

187

View full text Add to dashboard Cite

show abstract

“…The major immunoreactive protein was ϳ49 kDa. This is somewhat smaller than the predicted size of the full-length protein encoded by the PS1 cDNA (467 amino acids, M r 52,664; Sherrington et al, 1995), but it is similar to the M r of PS1 recently reported by others, using antibodies to the loop region and different epitopes at the N terminus (Elder et al, 1996;Mercken et al, 1996;Moussaoui et al, 1996;Thinakaran et al, 1996). Less intense immunoreactive bands of ϳ32 and ϳ26 kDa were also present in the PS1 translation products.…”

Section: Specificity Of a Monoclonal Antibody To The N Terminus Of Ps1supporting

confidence: 75%

“…Moreover, mutations in PS1 seem to alter the endoproteolytic processing, leading to an abnormal accumulation of the full-length protein Thinakaran et al, 1996). The light microscopic distribution of native PS1 in mouse brain (Elder et al, 1996;Moussaoui et al, 1996) and the localization of epitopetagged presenilins in transfected cells (Kovacs et al, 1996) have been reported. However, little is known about the presenilin proteins in primates.…”

Section: Abstract: Presenilin; Alzheimer's Disease; Immunocytochemismentioning

confidence: 99%

Light and Electron Microscopic Localization of Presenilin-1 in Primate Brain

et al. 1997

View full text Add to dashboard Cite

Several genes have been implicated in the pathogenesis of early-onset familial Alzheimer's disease. A majority of the autosomal dominant cases are linked to recently identified mutations in the presenilin-1 gene on chromosome 14. The native presenilin-1 protein in primates has not been well characterized, and its precise localization is unknown. We have studied the native presenilin-1 protein in monkey brain and peripheral tissues by using a monoclonal antibody specific for the N-terminal domain of human presenilin-1. Western blots detect polypeptide species of ϳ49 and ϳ32 kDa from COS-7 and PC12 cells transfected with full-length human presenilin-1 cDNA and from in vitro translations of the normal human presenilin-1 mRNA. A 32 kDa polypeptide is detected in monkey peripheral tissues, with the highest expression in testis and lung. In all brain regions the 32 kDa band is the predominant form of presenilin-1, and it is found in particulate subfractions. Light microscopic immunocytochemistry reveals presenilin-1 staining in all brain regions, with the strongest labeling in neurons and neuropil. In addition, weaker immunoreactivity is also present in glia and blood vessels. Neuronal staining shows significant variability, with particularly intense labeling of certain cell types, including large neocortical and hippocampal pyramidal neurons, magnocellular basal forebrain neurons, brainstem motoneurons, and some populations of interneurons. By electron microscopic immunocytochemistry, highly selective presenilin-1 staining is seen on the cytoplasmic surfaces of membranous organelles, which suggest localization to the endoplasmic reticulum-Golgi intermediate compartment, a subdomain of the endoplasmic reticulum, and some coated transport vesicles.

show abstract

“…First, message for PS2 showed a laminar distribution in cerebral cortex and was primarily detectable in neurons. This result is consistent with previous reports from normal human temporal lobes for PS2 expression visualized by radioactive hybridization [17] and is also comparable with PS1 expression pattern in murine brain observed using non-radioactive hybridization and immunohistochemistry [14,18]. Second, stronger hybridization signal was found in large neurons while mild or weak signal appeared in small neurons.…”

Section: Discussionsupporting

confidence: 92%

Gene expression of Alzheimer‐associated presenilin‐2 in the frontal cortex of Alzheimer and aged control brain

Deng

Cotman

1996

FEBS Letters

View full text Add to dashboard Cite

The presenilin-2 (PS2) gene expression pattern in AIzheimer's disease (AD) and control brains was examined using nonradioactive in situ hybridization. Message for PS2 was primarily detectable in neurons, particularly in somal cytoplasm. Intense staining signal was most commonly found in large pyramidal neurons, whereas moderate or faint staining was usually present in smaller neurons. The pattern of PS2 gene expression exhibited a laminar distribution proffie in the frontal cortex. A small subset of tangle-bearing neurons exhibited PS2 hybridization signal in AD. PS2 mRNA expression appeared correlated to a high degree with lipofuscin autofluorescence in a large subset of neurons.

show abstract

Immunohistochemical analysis of presenilin‐1 expression in the mouse brain

Cited by 56 publications

References 9 publications

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Light and Electron Microscopic Localization of Presenilin-1 in Primate Brain

Gene expression of Alzheimer‐associated presenilin‐2 in the frontal cortex of Alzheimer and aged control brain

Contact Info

Product

Resources

About