Intracellular and cell-surface distribution of amyloid precursor protein in cortical astrocytes.
Amyloid peptides that aggregate to form plaques in Alzheimer's disease are derived from secretory processing of the amyloid precursor protein (APP). Transport of APP to the cell surface may be prerequisite for non-amyloidogenic APP processing and the secretion of soluble APP (APPs), while missorting or reinternalization of APP to intracellular compartments can promote amyloid formation. In cultured astrocytes, APP mRNA and holoprotein are increased by elevations in cAMP levels, and 8-Bromo-cAMP promotes process formation on these cells. We now report that treatment of cultured astrocytes with 8-Bromo-cAMP increased intracellular and cell surface APP in the soma and perinuclear region as detected by immunolabeling with monoclonal antibody 22C11 and polyclonal antibody Kunitz-type protease inhibitor (KPI) (against the N-terminus and KPI domain of APP, respectively) and led to intense but discontinuous labelling of APP on the surface of astrocytic processes. Northern and Western blot analyses confirmed that 8-Bromo-cAMP treatment of cultured astrocytes also increased APP mRNA and KPI-containing APP holoprotein, implying that the intense APP immunolabeling observed in 8-Bromo-cAMP treated astrocytes was not derived from truncated forms of APP (e.g., APPs), but reflected high levels of APP holoprotein containing intact amyloid peptides. Discontinuous cell surface staining in process-bearing astrocytes may be caused by miscompartmentalization of APP related to rearrangement of the cytoskeleton. Inasmuch as intracellular APP is not accessible for non-amyloidogenic processing, we suggest that the increased immunoreactivity of intracellular APP in process-bearing astrocytes may predispose the cells to increased amyloid production.
Young
M J
MJ
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, USA. mikey@vision.eri.harvard.edu
Lee
R K
RK
Jhaveri
S
S
Wurtman
R J
RJ
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D016564:Amyloid beta-Protein Precursor / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal; D001253:Astrocytes / Q000737:chemistry* / Q000166:cytology / Q000502:physiology; D015152:Blotting, Northern; D015153:Blotting, Western; D002451:Cell Compartmentation / Q000502:physiology; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D000242:Cyclic AMP / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats
D016564:Amyloid beta-Protein Precursor; D000911:Antibodies, Monoclonal; D005904:Glial Fibrillary Acidic Protein; D008565:Membrane Proteins; D012333:RNA, Messenger; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP