Cloning of rat interleukin-3 receptor beta-subunit from cultured microglia and its mRNA expression in vivo

J Neurosci. 1995 Aug;15(8):5800-9. doi: 10.1523/JNEUROSCI.15-08-05800.1995.

Abstract

The high-affinity receptors for interleukin-3 (IL-3), GM-CSF, and IL-5 are composed of a ligand binding (alpha-) and a transducing (beta-) subunit. Two distinct transducing subunits (clones AIC2A and AIC2B) have been cloned from mouse, whereas in humans, only one (common) beta-subunit (beta c) has been found. A PCR-based cloning strategy was used to obtain a full-length cDNA sequence from rat microglia including 5'-untranslated regions. Sequence analysis revealed a number of features indicative of the presence of only one beta-subunit in the rat. Most likely, the new rIL-3R beta cDNA is the rat equivalent of human respective murine (AIC2B) beta c subunits. Regulation of rIL-3R beta mRNA expression was investigated in cultured microglia and in vivo. Purified microglia expressed significant amounts of rIL-3R beta mRNA. Addition of lipopolysaccharide (LPS) resulted in a marked upregulation of rIL-3R beta mRNA within approximately 4 hr. No downregulation was observed within 1 week's treatment. No rIL-3R beta mRNA was detectable in normal rat brain. However, 3 hr after a single injection of LPS into the tail vein of a rat, a marked induction of receptor mRNA occurred in a variety of brain regions. Transcriptional rates subsided significantly after 24 hr. rIL-3R beta mRNA was visualized by in situ hybridizations with cRNA antisense probes in ramified cells formerly characterized as microglial cells. rIL-3R beta mRNA was also induced in rat brain after occlusion of middle cerebral artery (MCAO).(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arterial Occlusive Diseases / metabolism
  • Base Sequence
  • Brain / cytology
  • Brain / metabolism
  • Cells, Cultured
  • Cerebral Arteries
  • Cloning, Molecular*
  • DNA, Complementary / genetics
  • Male
  • Microglia / metabolism*
  • Molecular Sequence Data
  • Oligonucleotide Probes / genetics
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Inbred SHR
  • Rats, Sprague-Dawley
  • Receptors, Interleukin-3 / genetics*
  • Recombinant Proteins

Substances

  • DNA, Complementary
  • Oligonucleotide Probes
  • RNA, Messenger
  • Receptors, Interleukin-3
  • Recombinant Proteins

Associated data

  • GENBANK/S79263