Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108‐15 cells

Gomes, Ivone; Gupta, Achla; Singh, Surya Pratap; Sharma, Sweta

doi:10.1016/s0014-5793(99)00878-9

Cited by 11 publications

(10 citation statements)

References 22 publications

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“…The band observed in the region of 60 -75 kDa most likely corresponds to the monomeric form of ␦OR. It is the closest to the size predicted from the amino acid sequence of the receptor and it is similar to that reported for the ␦OR in the same or different cell systems (Evans et al, 1992;Kieffer et al, 1992;Loukas et al, 1994;Standifer et al, 1995;Cvejic and Devi, 1997;Gomes et al, 1999;Ko et al, 1999). On the other hand, the heaviest band may well correspond to the 120-kDa ␦OR homodimer reported by Cvejic and Devi (1997).…”

Section: Specificity Of the Chemicon And Qcb ␦Or Antisera: Studies Insupporting

confidence: 88%

Immunohistochemical distribution of delta opioid receptors in the rat central nervous system: Evidence for somatodendritic labeling and antigen‐specific cellular compartmentalization

Cahill

McClellan

Morinville

et al. 2001

J of Comparative Neurology

131

129

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Many studies have reported on the distribution of delta opioid receptors (delta OR) in the mammalian central nervous system (CNS) by using a variety of techniques. However, no general consensus has emerged with regards to the localization of this receptor due to inconsistencies in the immunohistochemical literature. In the present study, we analyzed the cellular and subcellular distribution of immunoreactive delta OR in the rat CNS using two different antibodies (directed against a sequence in the C-terminus or N-terminus of the rat delta OR). By using Western blotting, these two antibodies recognized similar forms of the delta OR in COS-7 cells transfected with this receptor, but distinct forms in membranes from the rat spinal cord. By using light microscopic immunohistochemistry, both antibodies recognized identical populations of nerve cell bodies throughout the CNS; the distribution of these cell bodies conformed to that of delta OR mRNA-expressing cells detected by in situ hybridization. However, whereas the C-terminus-directed antibody recognized predominantly perikarya and proximal dendrites, the N-terminus-directed antibody also labeled extensively dendritic and terminal arbors. Furthermore, by using electron microscopy, the two antibodies were found not only to label differentially somatodendritic versus axonal compartments, but also plasma membrane versus cytoplasmic ones, suggesting that distinct immunological forms of the receptor are being targeted preferentially to different cellular and subcellular domains.

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Section: Specificity Of the Chemicon And Qcb ␦Or Antisera: Studies Insupporting

confidence: 88%

Immunohistochemical distribution of delta opioid receptors in the rat central nervous system: Evidence for somatodendritic labeling and antigen‐specific cellular compartmentalization

Cahill

McClellan

Morinville

et al. 2001

J of Comparative Neurology

131

129

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“…5C). However, in accord with previous reports, expression of the DOP subtype is predominant compared with the MOP (Gomes et al, 1999). It is noteworthy that in NG108-15 cells morphine transiently increased CRE activity after 4 h, but did not show a stimulatory effect after 16 h (Fig.…”

Section: Resultssupporting

confidence: 92%

“…Cell type-specific gene regulation by morphine raises an important question about the molecular determinants responsible for these differences. It is noteworthy that F11 cells express DOP and MOP but not opioid receptor subtypes (Fan et al, 1993), whereas NG108-15 cells almost exclusively express the DOP (Gomes et al, 1999). In line with this overall situation, the effects of DAMGO on cAMP production were much more pronounced in F11 compared with NG108-15 cells.…”

Section: Downloaded Fromsupporting

confidence: 55%

Morphine Activates the E Twenty Six-Like Transcription Factor-1/Serum Response Factor Pathway via Extracellular Signal-Regulated Kinases 1/2 in F11 Cells Derived from Dorsal Root Ganglia Neurons

Rothe

Solinski²,

Boekhoff³

et al. 2012

J Pharmacol Exp Ther

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Morphine-induced signaling via opioid receptors (ORs) in dorsal root ganglia (DRG) neurons, the spinal cord, and various brain regions has been shown to modulate gene activity. Hitherto, little attention has been paid to extracellular signal-regulated kinases-1/2 (ERK-1/2)-mediated activation of the serum response factor (SRF) and ternary complex factors (TCFs) such as the E twenty six-like transcription factor-1 (ELK-1) in this context. Using TCF/SRF-dependent reporter gene constructs, a specific ERK-1/2 inhibitor and a dominant-negative ELK-1 mutant, we show herein that morphine activates ELK-1 via ERK-1/2 in DRG-derived F11 cells endogenously expressing and ␦ ORs. Previous studies with glioma cell lines such as NG108-15 cells attributed morphine-induced gene expression to the activation of the cAMP-responsive element binding protein (CREB). Thus, we also analyzed morphine-dependent activation of CREB in F11 and NG108-15 cells. In contrast to the CREB stimulation found in NG108-15 cells, we observed an inhibitory effect of morphine in F11 cells, indicating cell typespecific regulation of CREB by morphine. To obtain data about putative target genes of morphine-induced ELK-1/SRF activation, we analyzed mRNA levels of 15 ELK-1/SRF-dependent genes in cultured rat DRG neurons and F11 cells. We identified the early growth response protein-4 (EGR-4) as the strongest up-regulated gene in both cell types and observed ELK-1 activity-dependent activation of an EGR-4-driven reporter in F11 cells. Overall, we reveal an important role of ELK-1 for morphine-dependent gene induction in DRG-derived cells and propose that ELK-1 and EGR-4 contribute to the effects of morphine on neuronal plasticity.

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“…Antisera to (SA25 and NT1), ␦ (LV17), ␣ 2A (GS29), or CB1 (DL41) MAPs were generated in rats and to ␤ 2 (SE27) or AT1 (LK12) MAPs in rabbits using a standard protocol (24). Monoclonal Abs to (5G8 and 3D6) MAPs were generated in mice as described previously (25). These antibodies are highly receptor-specific, since they exhibit low cross-reactivity against other closely related receptors, as examined using a whole cell ELISA (described below) with COS cells expressing the various receptors indicated above.…”

Section: Methodsmentioning

confidence: 99%

Conformation State-sensitive Antibodies to G-protein-coupled Receptors

Gupta

Décaillot

Gomes

et al. 2007

Journal of Biological Chemistry

Self Cite

118

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A growing body of evidence indicates that G-protein-coupled receptors undergo complex conformational changes upon agonist activation. It is likely that the extracellular region, including the N terminus, undergoes activation-dependent conformational changes. We examined this by generating antibodies to regions within the N terminus of -opioid receptors. We find that antibodies to the midportion of the N-terminal tail exhibit enhanced recognition of activated receptors, whereas those to the distal regions do not. The enhanced recognition is abolished upon treatment with agents that block G-protein coupling or deglycosylate the receptor. This suggests that the N-terminal region of receptors undergoes conformational changes following receptor activation that can be selectively detected by these regionspecific antibodies. We used these antibodies to characterize receptor type-specific ligands and find that the antibodies accurately differentiate ligands with varying efficacies. Next, we examined if these antibodies can be used to investigate the extent and duration of activation of endogenous receptors. We find that peripheral morphine administration leads to a time-dependent increase in antibody binding in the striatum and prefrontal cortex with a peak at about 30 min, indicating that these antibodies can be used to probe the spatio-temporal dynamics of native receptors. Finally, we show that this strategy of targeting the N-terminal region to generate receptor conformation-specific antisera can be applied to other G␣ i -coupled (␦-opioid, CB1 cannabinoid, ␣ 2A -adrenergic) as well as G␣ s -(␤ 2 -adrenergic) and G␣ q -coupled (AT1 angiotensin) receptors. Taken together, these studies describe antisera as tools that allow, for the first time, studies probing differential conformation states of G-protein-coupled receptors, which could be used to identify molecules of therapeutic interest.Family A GPCRs 2 play a critical role in normal cell function and are the focus of intense studies and targets for drug development. A tremendous effort has been put toward understanding the mechanism of activation of family A GPCRs at a molecular level. Spin label studies with rhodopsin have shown that exposure to light leads to a movement of helices relative to one another that is important for activation of transducin (1-4). Studies using a variety of techniques suggest that small agonists bind to a pocket formed by the surrounding transmembrane helices, and in addition peptide ligands contact additional determinants in extracellular loops and possibly the N-terminal tail (5-10). Binding of agonists, but not antagonists, leads to the stabilization of the helical bundle into a conformation, which, in turn, leads to the uncovering of molecular determinants at the bottom of the core required for G-protein binding and activation (11). Although a comprehensive mechanism for receptor activation, including the N-and C-terminal regions, is not yet available, accumulating evidence suggests that these regions undergo strong structural pertu...

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Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108‐15 cells

Cited by 11 publications

References 22 publications

Immunohistochemical distribution of delta opioid receptors in the rat central nervous system: Evidence for somatodendritic labeling and antigen‐specific cellular compartmentalization

Immunohistochemical distribution of delta opioid receptors in the rat central nervous system: Evidence for somatodendritic labeling and antigen‐specific cellular compartmentalization

Morphine Activates the E Twenty Six-Like Transcription Factor-1/Serum Response Factor Pathway via Extracellular Signal-Regulated Kinases 1/2 in F11 Cells Derived from Dorsal Root Ganglia Neurons

Conformation State-sensitive Antibodies to G-protein-coupled Receptors

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