Single-agent inhibition of wild-type and mutant muscle nAChRs
Cell-based study · Outside-out patches from HEK293 or BOSC23 cells expressing recombinant mouse adult muscle nAChRs (α2βεδ), wild type or αY198F/ε-subunit mutants. [1]
Pancuronium inhibited wild-type receptor current with a reported IC50 of 15 ± 1 nM. αY198F reduced potency, whereas four tested ε-subunit mutations did not significantly change IC50. These findings identify model-specific determinants of functional inhibition, not a direct structural binding measurement.
| Endpoint | Reported result | Reference |
|---|---|---|
| Equilibrium inhibition of ACh-evoked peak current in wild-type mouse adult muscle nAChR | IC50 15 ± 1 nM | [1] |
| Equilibrium inhibition in αY198F mutant compared with wild type | IC50 31 ± 8 nM; P < 0.001 versus wild type | [1] |
| Equilibrium inhibition in four ε-subunit mutants; no significant IC50 change reported | IC50 εD59A: 14 ± 2 nM; εD59N: 16 ± 3 nM; εD173A: 15 ± 3 nM; εD173N: 16 ± 2 nM | [1] |
Literature-Reported Treatment Conditions
- Treatment concentration
- Six or seven antagonist concentrations in concentration–response experiments; exact pancuronium series not enumerated in the read methods. Representative traces used 10 nM pancuronium. ACh challenge: 300 μM.
- Exposure duration
- Equilibrium protocol: 0.25-s ACh plus antagonist exposures separated by 5-s antagonist-only intervals; ten exposures in control protocol.
- Method
- Rapid perfusion, outside-out patch clamp at −50 mV and 20–23°C. Drug present before and during ACh application. Antagonist-free controls bracketed treatment; experiments accepted if second control amplitude was ≥80% of first.
- Vehicle
- Extracellular recording solution, pH 7.3; no separate pancuronium stock solvent specified in the read methods.
- Co-treatment
- Single antagonist with ACh as the assay agonist; other antagonists were tested separately.
- Reported limits
- Exact pancuronium concentration series and per-condition patch counts are not individually tabulated. Table 1 does not explicitly label the uncertainty type of its fitted IC50 ± values. No Adooq lot tested.
Methods and source locations
Compound reported in the paper: Pancuronium dibromide; Compound label in the paper: Pancuronium
Recombinant mouse adult muscle nicotinic acetylcholine receptors; experimental supplier material, not validation of Adooq material.
Materials and Methods; Results, Equilibrium Inhibition; Table 1; Figures 2C and 3
Equilibrium inhibition of ACh-evoked peak current in wild-type mouse adult muscle nAChR: Hill fit; Table 1 preserves the reported ± value without assigning an uncertainty label absent from that table. Methods report six or seven concentrations, each measured on three to six patches; plotted data are means and SDs. — Table 1, Pancuronium/Wild type; Materials and Methods, concentration–response analysis
Equilibrium inhibition in αY198F mutant compared with wild type: Hill fit; F test for mutant versus wild-type IC50. Reported ± retained without an inferred error type. — Table 1, Pancuronium/αY198F and dagger footnote; Figure 2C; Figure 3
Equilibrium inhibition in four ε-subunit mutants; no significant IC50 change reported: Hill fits; no significance marker for these pancuronium rows in Table 1; Discussion explicitly reports no effect of ε mutations. Reported ± retained without an inferred error type. — Table 1, Pancuronium ε-mutant rows; Figure 3; Discussion, mutation sensitivity