title: Cytoskeletal alterations in human fetal astrocytes induced by interleukin-1 beta.
abstract: Previous studies in this and other laboratories have shown that interleukin-1 beta (IL-1 beta) is a selective and potent activator of human astrocytes with respect to induction of cytokines and hematopoietic growth factors. To study the effect of recombinant human IL-1 beta (rhIL-1 beta) on astrocyte morphology, glial fibrillary acidic protein (GFAP) and vimentin expression, and actin organization, we conducted a systematic survey using dissociated human fetal astrocyte cultures. Within hours of stimulation with IL-1 beta, the majority of astrocytes converted from flat, polygonal cells to small, contracted, highly branched cells. This change in morphology was more striking when serum was eliminated from the medium. Complete dissolution of filamentous actin occurred simultaneously with the change in cell shape, as demonstrated by fluorescein-phalloidin binding. These "activated" astrocytes displayed intense GFAP and vimentin immunoreactivity in the small perikarya and processes. In contrast, the large, flat astrocytes in control cultures showed diffuse pale immunoreactivity for GFAP and vimentin. To quantify the changes in GFAP and vimentin content with IL-1 beta stimulation, densitometric analyses of northern and western blots were performed. Northern blot analysis of IL-1 beta-stimulated astrocytes revealed a transient, marked decrease in steady-state levels of mRNA for GFAP, vimentin, and microtubule-associated protein 4. The decrease in mRNA levels was evident by 4-8 h and fell to the lowest level at 16-24 h (80-98% decrease by densitometry) with partial recovery by 72 h. By immunoblotting, a significant decrease in both GFAP and vimentin protein content was observed after IL-1 beta stimulation. Furthermore, metabolic labeling studies revealed an almost total loss of GFAP synthesis following stimulation with IL-1 beta for 16 h. These observations are consistent with the idea that increases in immunoreactivity were related to factors such as redistribution of epitope, rather than increases in total protein content. We hypothesize that in IL-1 beta-stimulated astrocytes, synthesis of other proteins, e.g., inflammatory cytokines, occurs at the expense of structural proteins and that the decrease in content of cytoskeletal proteins may reflect an "activated" state of astrocytes.
authors.affiliation: Department of Pathology (Neuropathology), Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Shafit-Zagardo
authors.forename: D A
authors.initials: DA
authors.lastname: Aquino
authors.forename: M L
authors.initials: ML
authors.lastname: Zhao
authors.forename: D W
authors.initials: DW
authors.lastname: Dickson
authors.forename: C F
authors.initials: CF
authors.lastname: Brosnan
authors.forename: S C
authors.initials: SC
authors.lastname: Lee
mesh_terms: D001253:Astrocytes / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure*; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure*; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005333:Fetus / Q000166:cytology*; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D008869:Microtubule-Associated Proteins / Q000032:analysis; D011994:Recombinant Proteins / Q000494:pharmacology; D014746:Vimentin / Q000032:analysis
chemical_list: D005904:Glial Fibrillary Acidic Protein; D007375:Interleukin-1; D008869:Microtubule-Associated Proteins; D011994:Recombinant Proteins; D014746:Vimentin