Fentanyl Inhibits the Uptake of [ 3 H]noradrenaline in Cultured Neuronal Cells

Abstract

We have examined how fentanyl modulates [3H]noradrenaline uptake in two cultured neuronal cell preparations, the human neuroblastoma SH-SY5Y and the rat phaeochromocytoma PC12. Fentanyl produced a significant, dose-dependent inhibition of [3H]noradrenaline uptake at concentrations in excess of 0.1 mumol litre-1 (P < 0.05) and 0.3 mumol litre-1 (P < 0.05) for PC12 and SH-SY5Y cells, respectively. However, these values exceed the serum concentration of fentanyl required to produce analgesia. At the maximum concentration examined (100 mumol litre-1), fentanyl produced 85-95% inhibition of uptake. This effect was not antagonized by naloxone, implying a nonopioid mechanism of action. Imipramine 1 mumol litre-1 reduced [3H]noradrenaline uptake by 65-70% but morphine, in contrast to fentanyl, had no effect (P > 0.1).

Studies on the mechanism of [3H]‐noradrenaline release from SH‐SY5Y cells: the role of Ca2+ and cyclic AMP

“…Both tyrosine hydroxylase and dopamine-p-hydroxylase activities have been reported (Ross et al, 1983;Ross & Biedler, 1985) and these cells express muscarinic M3 receptors (Lambert et al, 1989), M2-adrenoceptors (Kazmi & Mishra, 1989) and ,p and 6 opioid receptors (Kasmi & Mishra, 1987). In a previous study we have shown that SH-SY5Y cells take up substantial amounts of [3H]-noradrenaline with a t4 of 5.7min (Atcheson et al, 1993b). Scott and colleagues (1986) demonstrated (in differentiated cells) that both K+ (50 mM) depolarization and acetylcholine (100 pLM) evoked a modest monophasic increase in the release of [3H]-noradrenaline; these data are confirmed and extended in this manuscript.…”

“…In addition, Seward et al (1991) demonstrated that opiate receptor occupancy in these cells closed VSCC. To address these discrepancies we have examined the roles played by intracel- Atcheson et al (1993b). Gross morphological examination showed no evidence of differentiation as a result of the experimental manipulations reported here.…”

“…In addition, Seward et al (1991) (Murphy et al, 1991;Atcheson et al, 1993b [3H]-noradrenaline-loaded cells were then pumped into the perfusion chamber (see below). The chamber was made from a 2 ml syringe barrel with the plunger replaced by a perspex flow spreader.…”

“…[3H]-noradrenaline release Loading of [3H]-noradrenaline and subsequent release from multiwell cultured cells was performed in Krebs/HEPES buffer supplemented with pargyline (0.2 mM) and ascorbic acid (0.2 mM) at 370C as described previously (Murphy et al, 1991;Atcheson et al, 1993b). Monolayers of SH-SY5Y cells were first rinsed with two 2 ml aliquots of Krebs/ HEPES buffer then loaded with 0.5 ml of [3H]-noradrenaline (48 nM in Krebs/HEPES buffer) for 1 h.…”

Noradrenergic Mechanisms in Fentanyl-Mediated Rapid Death Explain Failure of Naloxone in the Opioid Crisis

“…As previously described, fentanyl differs from morphine since the former has considerably increased lipid solubility, greater selectivity for m-opioid receptors over k and d, but has similar binding affinity as morphine on m-opioid receptors. In addition, fentanyl, but not morphine, is capable of acting as a norepinephrine reuptake inhibitor (Mather, 1983;Atcheson et al, 1993). High-dose fentanyl (e.g., 1-100 mM) blocks reuptake of [ 3 H]norepinephrine into human neuroblastoma and rat pheochromocytoma cells (Atcheson et al, 1993).…”