Cytoskeletal Alterations in Human Fetal Astrocytes Induced by Interleukin‐1β

Wei Liu, Bridget Shafit‐Zagardo, Dennis A. Aquino, Meng‐Liang ‐L Zhao, Dennis W. Dickson, Celia F. Brosnan, Sunhee C. Lee

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

Abstract: Previous studies in this and other laboratories have shown that interleukin‐1β (IL‐1β) 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‐1β (rhIL‐1β) 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‐1β, 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‐1β stimulation, densitometric analyses of northern and western blots were performed. Northern blot analysis of IL‐1β‐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‐1β stimulation. Furthermore, metabolic labeling studies revealed an almost total loss of GFAP synthesis following stimulation with IL‐1β 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‐1β‐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.

Original languageEnglish (US)
Pages (from-to)1625-1634
Number of pages10
JournalJournal of neurochemistry
Volume63
Issue number5
DOIs
StatePublished - Nov 1994

Keywords

  • Astrocytes
  • Cytokines
  • Cytoskeletal proteins
  • Interleukin‐1β

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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