Table 1.

Saturation binding of α2AAR and μOR in WT and NET-KO mouse spinal cord

WTNET-KO
BMAX (fmol/mg of protein)KD (nm)BMAX (fmol/mg of protein)KD (nm)
[3H]RX821002   193 ± 210.17 ± 0.01216 ± 210.28 ± 0.05
[3H]Prazosin76 ± 30.20 ± 0.01 35 ± 2**0.14 ± 0.03
[3H]Naloxone121 ± 7 2.3 ± 0.3112 ± 172.8 ± 0.4
  • Membranes were prepared from mouse spinal cords, and saturation binding curves were performed. The α2AAR antagonist [3H]RX821002 (0.2–10 nm), the α1AR antagonist [3H]prazosin (0.1–3 nm), and low concentrations of the opioid receptor antagonist [3H]naloxone (0.2–10 nm) were used to assess binding parameters of the respective receptors. Specific binding was determined by performing the experiments in the presence of either phentolamine (10 μm for α2AAR and 5 μm for α1AR) or naloxone (10 μm for μOR). Presented are binding parameters obtained from Scatchard analysis using the GraphPad Prism software. Data are the mean ± SEM of three experiments performed in duplicate.

  • **p < 0.001, versus WT, Student's t test.