Fentanyl increases intracellular Ca2+ concentrations in SH-SY5Y cells

Wandless, A L; Smart, Darren; Lambert, David G.

doi:10.1093/bja/76.3.461

Cited by 19 publications

(12 citation statements)

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“…Control experiments revealed that SH‐SY5Y cells produced an increase in [Ca 2+ ] i in response to the muscarinic receptor agonist carbachol (1 m M ). These data are consistent with our previous findings (Wandless et al ., 1996).…”

Section: Resultsmentioning

confidence: 96%

“…However we have previously shown that in SH‐SY5Y cells PLC activation is dependent upon Ca 2+ entry (Smart et al ., 1995). The subsequent Ins(1,4,5)P 3 formation does not appear to be sufficient to cause an increase in Ca 2+ , although μ‐opioid receptor activation in the same cells causes a small and unpredictable increase in [Ca 2+ ] i with very high concentrations of fentanyl (Wandless et al ., 1996), which may be due to calcium entry. Other studies have found that ‘priming’ of SH‐SY5Y cells with the muscarinic agonist carbachol is necessary to produce an opioid mediated increase in [Ca 2+ ] i (Connor & Henderson, 1996).…”

Section: Discussionmentioning

confidence: 99%

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The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

Harrison¹,

McNulty²,

Smart³

et al. 1999

British J Pharmacology

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1 Endomorphin-1 and -2 (E-1/E-2) have been proposed as endogenous ligands for the m-opioid receptor. The aims of this study are to characterize the binding of E-1/E-2 and the subsequent e ects on cyclic AMP formation and 2+ ] i in fura-2 loaded CHOm cell suspensions in a thapsigargin sensitive and naloxone reversible manner. Mean increases observed were 106+28 and 69+6.7 nM respectively. In single adherent cells E-1/E-2 (1 mM) increased [Ca 2+ ] i with a mean 340/380 ratio change of 0.81+0.09 and 0.40+0.08 ratio units respectively. E-1/E-2 failed to increase intracellular calcium in CHOd, CHOk and SH-SY5Y cells. 5 These data show that E-1/E-2 bind with high a nity and selectivity to m-opioid receptors and modulate signal transduction pathways typical of opioids. This provides further evidence that these two peptides may be endogenous ligands at the m-opioid receptor.

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Section: Resultsmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

Harrison¹,

McNulty²,

Smart³

et al. 1999

British J Pharmacology

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“…Although morphine induced less pCREB formation after cellular depletion of PKC by PMA pretreatment, this increase was abolished by KN62, arguing for a role of both PKC and CaMKs. In SH‐SY5Y cells, opioid receptor stimulation leads to activation of PKC (Smart et al, 1995; Wandless et al, 1996). Whether PKC plays a direct role in CREB phosphorylation remains to be seen.…”

Section: Discussionmentioning

confidence: 99%

“…These signaling pathways suppress neuronal excitability; however, opioid receptors also trigger various poorly defined signaling pathways that can stimulate cellular activity. For example, opioid receptors have been shown to enhance Ca 2+ influx/release (Smart et al, 1995; Wandless et al, 1996; Thorlin et al, 1998; Lin et al, 1999) and to activate mitogen‐activated protein kinase (MAPK) (Fukuda et al, 1996; Hawes et al, 1998) via G i /G o proteins.…”

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confidence: 99%

Nuclear Ca²⁺/Calmodulin Translocation Activated by μ‐Opioid(OP₃) Receptor

Wang

Tolbert²,

Carlson³

et al. 2000

Journal of Neurochemistry

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Abstract:Previous evidence has suggested a role for calmodulin (CaM) in opioid receptor signaling. We demonstrate here that morphine stimulation of the -opioid (OP 3 ) receptor causes rapid CaM translocation to the nucleus in OP 3 -transfected human embryonic kidney (HEK)-293 cells and in SH-SY5Y human neuroblastoma cells. Ca 2ϩ influx into the cells resulting from OP 3 receptor activation was required for nuclear CaM translocation. Moreover, in HEK-OP 3 and SH-SY5Y cells, increased nuclear CaM content was associated with enhanced phosphorylation of the nuclear transcription factor cyclic AMP-responsive element-binding protein. This appeared to be mediated by Ca 2ϩ /CaM kinases and also by a pathway involving protein kinase C. CaM was previously shown to bind directly to the OP 3 receptor and to be released from the plasma membrane on agonist stimulation. To test whether OP 3 -mediated CaM release contributes to nuclear CaM signaling, we used a mutant OP 3 receptor (K273A) with reduced affinity for CaM that fails to release CaM from the plasma membrane. K273A-OP 3 activated Ca 2ϩ influx to a similar extent as wild-type OP 3 ; however, CaM translocation to the nucleus was attenuated. These results indicate that OP 3 -stimulated Ca 2ϩ influx results in nuclear CaM translocation, which appears to be enhanced by simultaneous CaM release by OP 3 wild-type receptor from plasma membranes. These results suggest a novel Ca 2ϩ /CaM signaling pathway of opioid receptors in the regulation of transcriptional activity.

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“…Endogenous μ‐opioid receptors in SH‐SY5Y human neu‐roblastoma cells also inhibit N‐type VSCCs (Seward et al , 1991), as well as basal and forskolin‐stimulated cyclicAMP formation (Lambert et al , 1993; Smart et al , 1995a). Furthermore, the μ‐opioid receptor also stimulates Ins(1,4,5)P 3 formation in these cells (Smart et al , 1994; 1995a), by opening L‐type VSCCs (Smart & Lambert, 1995; Smart et al , 1995a) via a pertussis toxin (PTX)‐sensitive G‐protein (Smart et al , 1994), and so allowing Ca 2+ influx (Wandless et al , 1996) which activates PLC.…”

Section: Introductionmentioning

confidence: 99%

The effects of recombinant rat μ‐opioid receptor activation in CHO cells on phospholipase C, [Ca²⁺]_i and adenylyl cyclase

Smart

Hirst

Hirota

et al. 1997

British J Pharmacology

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1. The rat mu-opioid receptor has recently been cloned yet its second messenger coupling remains unclear. The endogenous mu-opioid receptor in SH-SY5Y cells couples to phospholipase C (PLC), increases [Ca2+]i and inhibits adenylyl cyclase (AC). We have examined the effects of mu-opioid agonists on inositol(1,4,5)trisphosphate (Ins(1,4,5)P3), [Ca2+]i and adenosine 3':5'-cyclic monophosphate (cyclic AMP) formation in Chinese hamster ovarian (CHO) cells transfected with the cloned mu-opioid receptor. 2. Opioid receptor binding was assessed with [3H]-diprenorphine ([3H]-DPN) as a radiolabel. Ins(1,4,5)P3 and cyclic AMP were measured by specific radioreceptor assays. [Ca2+]i was measured fluorimetrically with Fura-2. 3. Scatchard analysis of [3H]-DPN binding revealed that the Bmax varied between passages. Fentanyl (10 pM 1 microM) dose-dependently displaced [3H]-DPN, yielding a curve which had a Hill slope of less than unity (0.6 +/- 0.1), and was best fit to a two site model, with pK1 values (% of sites) of 9.97 +/- 0.4 (27 +/- 4.8%) and 7.68 +/- 0.07 (73 +/- 4.8%). In the presence of GppNHp (100 microM) and Na+ (100 mM), the curve was shifted to the right and became steeper (Hill slope = 0.9 +/- 0.1) with a pK1 value of 6.76 +/- 0.04. 4. Fentanyl (0.1 nM-1 microM) had no effect on basal, but dose-dependently inhibited forskolin (1 microM)-stimulated, cyclic AMP formation (pIC50 -7.42 +/- 0.23), in a pertussis toxin (PTX; 100 ng ml-1 for 24 h)-sensitive and naloxone-reversible manner (K1 = 1.7 nM). Morphine (1 microM) and [D-Ala2, MePhe4, gly(ol)5]-enkephalin (DAMGO, 1 microM) also inhibited forskolin (1 microM)-stimulated cyclic AMP formation, whilst [D-Pen2, D-Pen5], enkephalin (DPDPE, 1 microM) did not. 5. Fentanyl (0.1 nM-10 microM) caused a naloxone (1 microM)-reversible, dose-dependent stimulation of Ins(1,4,5)P3 formation, with a pEC50 of 7.95 +/- 0.15 (n-5), PTX (100 ng ml-1 for 24 h) abolished, whilst Ni2 (2.5 mM) inhibited (by 52%), the fentanyl-induced Ins(1,4,5)P3 response. Morphine (1 microM) and DAMGO (1 microM), but not DPDPE (1 microM), also stimulated Ins(1,4,5)P3 formation. Fentanyl (1 microM) also caused an increase in [Ca2+]i (80 +/- 16.4 nM, n-6), reaching a maximum at 26.8 +/- 2.5 s. The increase in [Ca2+]i remained elevated until sampling ended (200 s) and was essentially abolished by the addition of naloxone (1 microM). Pre-incubation with naloxone (1 microM, 3 min) completely abolished fentanyl-induced increases in [Ca2+]i. 6. In conclusion, the cloned mu-opioid receptor when expressed in CHO cells stimulates PLC and inhibits AC, both effects being mediated by a PTX-sensitive G-protein. In addition, the receptor couples to an increase in [Ca2+]i. These findings are consistent with the previously described effector-second messenger coupling of the endogenous mu-opioid receptor.

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Fentanyl increases intracellular Ca2+ concentrations in SH-SY5Y cells

Cited by 19 publications

References 5 publications

The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

Nuclear Ca²⁺/Calmodulin Translocation Activated by μ‐Opioid(OP₃) Receptor

The effects of recombinant rat μ‐opioid receptor activation in CHO cells on phospholipase C, [Ca²⁺]_i and adenylyl cyclase

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Fentanyl increases intracellular Ca2+ concentrations in SH-SY5Y cells

Cited by 19 publications

References 5 publications

The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

The effects of endomorphin‐1 and endomorphin‐2 in CHO cells expressing recombinant μ‐opioid receptors and SH‐SY5Y cells

Nuclear Ca2+/Calmodulin Translocation Activated by μ‐Opioid(OP3) Receptor

The effects of recombinant rat μ‐opioid receptor activation in CHO cells on phospholipase C, [Ca2+]i and adenylyl cyclase

Contact Info

Product

Resources

About

Nuclear Ca²⁺/Calmodulin Translocation Activated by μ‐Opioid(OP₃) Receptor

The effects of recombinant rat μ‐opioid receptor activation in CHO cells on phospholipase C, [Ca²⁺]_i and adenylyl cyclase