Degradation of glial glutamate transporter mRNAs is selectively blocked by inhibition of cellular transcription

J Neurochem. 2000 Dec;75(6):2252-8. doi: 10.1046/j.1471-4159.2000.0752252.x.

Abstract

Recent studies have demonstrated that the expression of the glial glutamate transporters GLT-1 (glutamate transporter 1) and GLAST (glutamate aspartate transporter) is regulated both in vivo and in vitro. For example, co-culturing with neurons, treatment with N:(6), 2'-O:-dibutyryladenosine 3':5'-cyclic monophosphate (dbcAMP), and treatment with epidermal growth factor all increase the steady-state levels of GLT-1 and GLAST protein in astrocyte cultures. These changes in protein expression are correlated with increased mRNA levels. In the present study, the degradation of GLT-1 and GLAST mRNAs was examined in control and dbcAMP-treated astrocyte cultures after inhibiting transcription with actinomycin D. Although one would predict that inhibition of transcription would cause a decrease in GLT-1 and GLAST mRNAs and that this decrease would depend on the rate of mRNA degradation, the levels of GLT-1 and GLAST mRNAs did not decrease even after 24 h of treatment with actinomycin D. Withdrawal of dbcAMP caused the levels of GLT-1 and GLAST mRNAs to fall to basal levels within 24 h, but this degradation was blocked if actinomycin D was added at the time of dbcAMP withdrawal. Importantly, actinomycin D did not block the degradation of c-fos mRNA also induced by dbcAMP in these cultures. Inhibition of translation with cycloheximide did not stabilize GLT-1 but partially attenuated the degradation of GLAST mRNA. Although the mechanism of this effect remains to be defined, these studies suggest that GLT-1 and GLAST mRNAs belong to a select class of inducible mRNAs stabilized by inhibitors of transcription. The possible relevance of these data to astrocyte differentiation is briefly discussed.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / metabolism*
  • Amino Acid Transport System X-AG
  • Animals
  • Astrocytes / cytology
  • Astrocytes / drug effects
  • Astrocytes / metabolism
  • Bucladesine / pharmacology
  • Cells, Cultured
  • Cycloheximide / pharmacology
  • Dactinomycin / pharmacology
  • Protein Synthesis Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-fos / genetics
  • Proto-Oncogene Proteins c-fos / metabolism
  • RNA Processing, Post-Transcriptional / drug effects
  • RNA Processing, Post-Transcriptional / genetics*
  • RNA, Messenger / metabolism*
  • Rats
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / genetics*

Substances

  • ATP-Binding Cassette Transporters
  • Amino Acid Transport System X-AG
  • Protein Synthesis Inhibitors
  • Proto-Oncogene Proteins c-fos
  • RNA, Messenger
  • Dactinomycin
  • Bucladesine
  • Cycloheximide