Glial-restricted precursors: patterns of expression of opioid receptors and relationship to human immunodeficiency virus-1 Tat and morphine susceptibility in vitro

Neuroscience. 2007 Jun 8;146(4):1546-54. doi: 10.1016/j.neuroscience.2007.03.006. Epub 2007 May 2.


Recent evidence suggests that human immunodeficiency virus (HIV)-induced pathogenesis is exacerbated by opioid abuse and that the synergistic toxicity may result from direct actions of opioids in immature glia or glial precursors. To assess whether opioids and HIV proteins are directly toxic to glial-restricted precursors (GRPs), we isolated neural stem cells from the incipient spinal cord of embryonic day 10.5 ICR mice. GRPs were characterized immunocytochemically and by reverse transcriptase-polymerase chain reaction (RT-PCR). At 1 day in vitro (DIV), GRPs failed to express mu opioid receptors (MOR or MOP) or kappa-opioid receptors (KOR or KOP); however, at 5 DIV, most GRPs expressed MOR and KOR. The effects of morphine (500 nM) and/or Tat (100 nM) on GRP viability were assessed in GRPs at 5 DIV by examining the apoptotic effector caspase-3 and cell viability (ethidium monoazide exclusion) at 96 h following continuous exposure. Tat or morphine alone or in combination caused significant increases in GRP cell death at 96 h, but not at 24 h, following exposure. Although morphine or Tat caused increases in caspase-3 activity at 4 h, this was not accompanied with increased cleaved caspase-3 immunoreactive or ethidium monoazide-positive dying cells at 24 h. The results indicate that prolonged morphine or Tat exposure is intrinsically toxic to isolated GRPs and/or their progeny in vitro. Moreover, MOR and KOR are widely expressed by Sox2 and/or Nkx2.2-positive GRPs in vitro and the pattern of receptor expression appears to be developmentally regulated. The temporal requirement for prolonged morphine and HIV-1 Tat exposure to evoke toxicity in glia may coincide with the attainment of a particular stage of maturation and/or the development of particular apoptotic effector pathways and may be unique to spinal cord GRPs. Should similar patterns occur in vivo then we predict that immature astroglia and oligodendroglia may be preferentially vulnerable to HIV-1 infection or chronic opiate exposure.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caspase 3 / metabolism
  • Cell Survival / drug effects
  • Cells, Cultured
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Embryo, Mammalian
  • Embryonic Stem Cells / drug effects*
  • Embryonic Stem Cells / physiology
  • Female
  • Gene Expression Regulation / drug effects
  • Gene Products, tat / pharmacology*
  • Homeobox Protein Nkx-2.2
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Mice
  • Mice, Inbred ICR
  • Morphine / pharmacology*
  • Narcotics / pharmacology*
  • Neuroglia / drug effects*
  • Neuroglia / physiology
  • Nuclear Proteins
  • Pregnancy
  • RNA, Messenger / biosynthesis
  • Receptors, Opioid / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • SOXB1 Transcription Factors
  • Spinal Cord / cytology
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Zebrafish Proteins
  • tat Gene Products, Human Immunodeficiency Virus


  • DNA-Binding Proteins
  • Gene Products, tat
  • Homeobox Protein Nkx-2.2
  • Homeodomain Proteins
  • NKX2-2 protein, human
  • Narcotics
  • Nkx2-2 protein, mouse
  • Nuclear Proteins
  • RNA, Messenger
  • Receptors, Opioid
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Trans-Activators
  • Transcription Factors
  • Zebrafish Proteins
  • nkx2.2b protein, zebrafish
  • tat Gene Products, Human Immunodeficiency Virus
  • tat peptide (1-72), Human immunodeficiency virus 1
  • Morphine
  • Caspase 3