Detection of cytokine mRNA in astrocyte cultures using the polymerase chain reaction

S. L. Wesselingh, N. M. Gough, J. J. Finlay-Jones, P. J. McDonald

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105 Citations (Scopus)

Abstract

To clarify which cytokines could potentially be produced by astrocytes, we have assessed the presence of mRNA for a number of cytokines in astrocyte-enriched brain cultures. The cultures were derived from neonatal murine brain and were treated with either interferon-γ, lipopolysaccharide or tumor necrosis factor-α, or infected with murine cytomegalovirus. RNA was extracted at 0, 4, 24 and 48 hours post treatment. A DNA copy of total cytoplasmic RNA was synthesised and specific cDNA amplified using the polymerase chain reaction and detected by hybridization with specific probes. The following cytokines were studied: IL3, IL4, IL6, TNFα, TGFβ, LIF, G-CSF, M-CSF and GM-CSF. Transcripts for TGFβ, IL6, LIF and M-CSF were present consitutively but increased with stimulation, whereas transcripts for TNFα, IL6, GM-CSF, G-CSF and LIF were detected only after stimulation. Messenger RNA for IL3 and IL4 was not detected. The magnitude and kinetics of the induction varied according to the cytokine and the stimulus. These results indicate the possibility of intra-cerebral production of a number of cytokines that may play a role in the clearance of viral or bacterial pathogens and in the generation of neuropathology.

LanguageEnglish
Pages177-185
Number of pages9
JournalLymphokine Research
Volume9
Issue number2
Publication statusPublished - 1990
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Hematology

Cite this

Wesselingh, S. L., Gough, N. M., Finlay-Jones, J. J., & McDonald, P. J. (1990). Detection of cytokine mRNA in astrocyte cultures using the polymerase chain reaction. Lymphokine Research, 9(2), 177-185.