Human μ-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins

Wei, Qiang; Zhou, De He; Shen, Qing; Chen, Jie; Chen, Li Wei; Wang, Tie Lin; Pei, Gang; Qiang, Zhi

doi:10.1038/sj.cr.7290039

Cited by 13 publications

(9 citation statements)

References 18 publications

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“…Since the human A-opioid receptors expressed in Sf9 cells showed a pharmacological profile similar to those described in mammalian cells (Massotte et al, 1997(Massotte et al, , 1999, the baculovirus/insect cell system provides a viable alternative for large receptor production. We have previously expressed human A-opioid receptors in Sf9 cells and demonstrated that these recombinant A-opioid receptors were functionally coupled to the endogenous G i/o proteins (Wei et al, 1998(Wei et al, , 2000. It provides a cellular model to study the effect of opiates.…”

Section: Discussionmentioning

confidence: 96%

“…Infection was performed in monolayer at a cell density of 2 Â 10 6 by recombinant baculoviruses containing human A-opioid receptor cDNA as previously described (Wei et al, 1998(Wei et al, , 2000. Cells were harvested at the peak time of production (60-64 h after the infection).…”

Section: Cell Culturementioning

confidence: 99%

“…Acute exposure of these Sf9-A cells with A agonists led to inhibition of forskolin-stimulated cAMP accumulation (Wei et al, 2000). After chronic pretreated Sf9-A cells with some opiates such as morphine, fentanyl and heroin, addition of naloxone induced intracellular cAMP overshoot (Liu et al, 2003).…”

Section: Introductionmentioning

confidence: 97%

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Effect of chronic treatment of ohmefentanyl stereoisomers on cyclic AMP formation in Sf9 insect cells expressing human μ-opioid receptors

Jin

et al. 2004

Life Sciences

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Section: Discussionmentioning

confidence: 96%

Section: Cell Culturementioning

confidence: 99%

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Effect of chronic treatment of ohmefentanyl stereoisomers on cyclic AMP formation in Sf9 insect cells expressing human μ-opioid receptors

Jin

et al. 2004

Life Sciences

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“…On the other hand, previous functional studies of a number of human GPCRs transiently expressed in both Sf9 and High Five™ cell lines showed agonist responses either leading to an elevation of intracellular Ca2+ concentrations (Kuhn et al, 1996) or a decrease in intracellular cAMP levels (Jansson et al, 1995;Obermeier et al, 1996;Wei et al, 2000), implying the presence of endogenous insect Gαq-like and Gαi-like α subunits, respectively. This is in full accord with our data.…”

Section: Archives Of Insect Biochemistry and Physiologymentioning

confidence: 94%

Diversity of G proteins in Lepidopteran cell lines: Partial sequences of six G protein alpha subunits

Knight

Grigliatti

2004

Arch Insect Biochem Physiol

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The aim of this work was to sample the diversity of G protein alpha subunits in lepidopteran insect cell lines. Here we report the amplification by degenerate PCR of partial sequences representing six G protein alpha subunits from three different lepidopteran insect cell lines. Sequence comparisons with known G protein alpha subunits indicate that the Sf9, Ld and High Five cell lines each contain (at least) one Galpha(q)-like and one Galpha(i)-like Galpha subunit. All six PCR products are unique at the nucleotide level, but the translation products of the three Galpha q-like partial clones (Sf9-Galpha 1, Ld-Galpha 1, and Hi5-Galpha 1) are identical, as are the translation products of the three Galpha i-like partial clones (Sf9-Galpha 2, Ld-Galpha 2, and Hi5-Galpha 2). Both the Galpha(q)-like and Galpha(i)-like translation products are identical to known Galpha subunits from other Lepidoptera, are highly similar (88-98%) to Galpha subunits from other invertebrates including mosquitoes, fruit flies, lobsters, crabs, and snails, and are also highly similar (88-90%) to known mammalian Galpha subunits. Identification of G protein alpha subunits in lepidopteran cell lines will assist in host cell line selection when insect cell lines are used for the pharmacological analysis of human GPCRs.

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“…The effect of F9204 was similar to morphine which effect was more potent and longer than that of F9202. Ohmefentanyl which is the most potent analgesic so far [28], has 8 isomers (F9201-F9208). F9202 and F9204 had similarly potent analgesic action, but showed a large difference in physical dependence [23].…”

Section: Discussionmentioning

confidence: 99%

Ohmefentanyl stereoisomers induce changes of CREB phosphorylation in hippocampus of mice in conditioned place preference paradigm

Gao

Chen

et al. 2003

Cell Res

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The present study was designed to determine the changes of phosphorylation of cAMP-response element binding protein (CREB) in hippocampus induced by ohmefentanyl stereoisomers (F9202 and F9204) in conditioned place preference (CPP) paradigm. The results showed that mice receiving F9202 and F9204 displayed obvious CPP. They could all significantly stimulate CREB phosphorylation and maintained for a long time without affecting total CREB protein levels. The effect of F9204 was similar to morphine which effect was more potent and longer than F9202. We also examined the effects of ketamine, a noncompetitive N-mthyl-D-aspartate receptor (NR) antagonist, on morphine-, F9202-and F9204-induced CPP and phosphorylation of CREB in hippocampus. Ketamine could suppress not only the place preference but also the phosphorylation of CREB produced by morphine, F9202 and F9204. These findings suggest that alterations in the phosphorylation of CREB be relevant to opiates signaling and the development of opiates dependence. NR antagonists may interfere with opiates dependence and may have potential therapeutic implications.

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Human μ-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins

Abstract: Human m-opioid receptor (H OR) with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells.

Cited by 13 publications

References 18 publications

Effect of chronic treatment of ohmefentanyl stereoisomers on cyclic AMP formation in Sf9 insect cells expressing human μ-opioid receptors

Effect of chronic treatment of ohmefentanyl stereoisomers on cyclic AMP formation in Sf9 insect cells expressing human μ-opioid receptors

Diversity of G proteins in Lepidopteran cell lines: Partial sequences of six G protein alpha subunits

Ohmefentanyl stereoisomers induce changes of CREB phosphorylation in hippocampus of mice in conditioned place preference paradigm

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