Basic fibroblast growth factor-2 and interleukin-1 beta regulate S100 beta expression in cultured astrocytes.
Basic fibroblast growth factor and interleukin-1 beta are known to regulate the expression of other trophic factors and to stimulate reactive gliosis in vivo. S100 beta is a glial-specific putative neurotrophic factor and has been considered a marker of the reactive status of astrocytes. Therefore, we tested the hypothesis that basic fibroblast growth factor-2 and interleukin-1 beta achieve their effects by altering S100 beta gene expression in cultured rat astrocytes using an RNase protection assay. Short-term treatment with basic fibroblast growth factor-2 produced a transient decrease in S100 beta messenger RNA which was followed by an increase after longer term treatment. In contrast, both short- and long-term treatment with interleukin-1 beta suppressed S100 beta messenger RNA. We measured levels of S100 beta nuclear primary transcript to assess whether alterations in transcriptional rate explain the changes in messenger RNA. Our results indicate that changes in transcription account for changes in steady state levels of messenger RNA since basic fibroblast growth factor-2-induced changes in S100 beta primary transcript temporally preceded changes in messenger RNA. We further measured intracellular S100 beta protein levels by enzyme-linked immunosorbent assay to determine whether changes in gene expression were translated into parallel changes in protein. Our results clearly demonstrate that basic fibroblast growth factor-2 and interleukin-1 beta influence the expression of the S100 beta gene, that this regulation appears to occur at the level of transcription, and that alterations in messenger RNA are sometimes, but not always, reflected in changes at the level of protein. These observations suggest that basic fibroblast growth factor-2 may amplify its trophic effects, in part, by influencing the expression of another trophic factor.
Hinkle
D A
DA
Department of Physiology, University of Maryland School of Medicine, Baltimore 21202, USA.
Harney
J P
JP
Cai
A
A
Hilt
D C
DC
Yarowsky
P J
PJ
Wise
P M
PM
D000818:Animals; D001253:Astrocytes / Q000378:metabolism* / Q000648:ultrastructure; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis*; D015870:Gene Expression / Q000502:physiology; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000096:biosynthesis*; D012313:RNA / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012260:Ribonucleases / Q000378:metabolism; D009418:S100 Proteins / Q000096:biosynthesis
D007375:Interleukin-1; D009418:S100 Proteins; D016222:Fibroblast Growth Factor 2; D012313:RNA; D012260:Ribonucleases