scholarly journals Investigating the Effect of Kappa Opioid Receptor Agonists on Serotonin Transporter Function in HEK-293 Cells

2021 ◽  
Author(s):  
◽  
Victoria Catherwood
Keyword(s):  
Opioid Receptor ◽  
High Throughput ◽  
Cell Model ◽  
Kappa Opioid Receptor ◽  
Rotating Disc ◽  
Kappa Opioid ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
High Throughput Assay

<p>The serotonin (5-HT) transporter (SERT) and the kappa opioid receptor (KOPr) are important brain proteins. Currently, it is known that KOPr modulates 5-HT concentrations but the biochemical mechanisms responsible remain enigmatic. The 5-HT and KOPr relationship is thought to involve SERT because both SERT and KOPr co-localise in the dorsal raphe nucleus (DRN), a brain region involved in drug addiction, affective disorders, and stress homeostasis. Thus, elucidating the KOPr and SERT relationship will clarify biomedical targets for these disorders. To investigate this relationship the effect of KOPr activation on SERT function was examined in HEK-293 cells co-expressing myc-rKOPr and eGFP-tagged hSERT. We found the KOPr agonists U50,488H, Salvinorin A (Sal A) and DS-3-240 had no acute (5 min) effect on SERT function as measured by rotating disc electrode voltammetry (RDEV) incubated with 1’-diethyl-2,2’-cyanine iodide (D-22; 1.5 mM) and by confocal microscopy. Interestingly, SERT function significantly decreased with chronic (30 min) exposure to U50,488H (**p<0.01) and Sal A (**p<0.01) as measured with a novel high-throughput assay; this decrease was also attenuated with 5 min pre-treatment of the SERT inhibitor, fluoxitene (***p<0.001). Furthermore, this novel high-throughput assay also replicated our laboratories recent finding that chronic (30 min) exposure to U50,488H (**p<0.01) and Sal A (**p<0.01) significantly increase dopamine transporter (DAT) function in HEK-293 cells co-expressing myc-rKOPr and eYFP-tagged DAT. Collectively, these findings suggest in this cell model that chronic (30 min) KOPr activation by U50,488H and Sal A decreases SERT function.</p>

scholarly journals Investigating the Effect of Kappa Opioid Receptor Agonists on Serotonin Transporter Function in HEK-293 Cells

2021 ◽  
Author(s):  
◽  
Victoria Catherwood
Keyword(s):  
Opioid Receptor ◽  
High Throughput ◽  
Cell Model ◽  
Kappa Opioid Receptor ◽  
Rotating Disc ◽  
Kappa Opioid ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
High Throughput Assay

<p>The serotonin (5-HT) transporter (SERT) and the kappa opioid receptor (KOPr) are important brain proteins. Currently, it is known that KOPr modulates 5-HT concentrations but the biochemical mechanisms responsible remain enigmatic. The 5-HT and KOPr relationship is thought to involve SERT because both SERT and KOPr co-localise in the dorsal raphe nucleus (DRN), a brain region involved in drug addiction, affective disorders, and stress homeostasis. Thus, elucidating the KOPr and SERT relationship will clarify biomedical targets for these disorders. To investigate this relationship the effect of KOPr activation on SERT function was examined in HEK-293 cells co-expressing myc-rKOPr and eGFP-tagged hSERT. We found the KOPr agonists U50,488H, Salvinorin A (Sal A) and DS-3-240 had no acute (5 min) effect on SERT function as measured by rotating disc electrode voltammetry (RDEV) incubated with 1’-diethyl-2,2’-cyanine iodide (D-22; 1.5 mM) and by confocal microscopy. Interestingly, SERT function significantly decreased with chronic (30 min) exposure to U50,488H (**p<0.01) and Sal A (**p<0.01) as measured with a novel high-throughput assay; this decrease was also attenuated with 5 min pre-treatment of the SERT inhibitor, fluoxitene (***p<0.001). Furthermore, this novel high-throughput assay also replicated our laboratories recent finding that chronic (30 min) exposure to U50,488H (**p<0.01) and Sal A (**p<0.01) significantly increase dopamine transporter (DAT) function in HEK-293 cells co-expressing myc-rKOPr and eYFP-tagged DAT. Collectively, these findings suggest in this cell model that chronic (30 min) KOPr activation by U50,488H and Sal A decreases SERT function.</p>

Phosphorylation and Agonist-Specific Intracellular Trafficking of an Epitope-Tagged μ-Opioid Receptor Expressed in HEK 293 Cells

2002 ◽  
Vol 65 (4) ◽  
pp. 1636-1645 ◽  
Author(s):  
James R. Arden ◽  
Veronica Segredo ◽  
Zaijie Wang ◽  
Jelveh Lameh ◽  
Wolfgang Sadée
Keyword(s):  
Opioid Receptor ◽  
Intracellular Trafficking ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Μ Opioid Receptor

scholarly journals Curcumin Antagonizes Glucose Fluctuation-Induced Renal Injury by Inhibiting Aerobic Glycolysis via the miR-489/LDHA Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-25
Author(s):  
Xiaomei Fu ◽  
Jianfang Zhang ◽  
Xuanjie Huang ◽  
Zhifeng Mo ◽  
Ziyang Sang ◽  
...  
Keyword(s):  
Renal Injury ◽  
Warburg Effect ◽  
Cell Model ◽  
Aerobic Glycolysis ◽  
Glucose Fluctuation ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
A Cell ◽  
The Warburg Effect

It has been considered that glucose fluctuation (GF) plays a role in renal injury and is related to diabetic nephropathy (DN) development. But the mechanism is still unclear. Aerobic glycolysis has become a topical issue in DN in recent years. There is an internal connection between GF, aerobic glycolysis, and DN. Curcumin (Cur) is a principal curcuminoid of turmeric and possesses specific protective properties in kidney functions. Cur also participates in the regulation of aerobic glycolysis switch. In this study, we first measured the levels of aerobic glycolysis and evaluated Cur’s inhibitory ability in a cell model of HEK-293 under the condition of oscillating high glucose. The results indicated that GF exacerbated inflammation injury, oxidative stress, and apoptosis in HEK-293 cell, while Cur alleviated this cytotoxicity induced by GF. We found that GF increased aerobic glycolysis in HEK-293 cells and Cur presented a dose-dependent weakening effect to this exacerbation. Next, we built a panel of 17 miRNAs and 8 lncRNAs that were previously reported to mediate the Warburg effect. Our RT-qPCR results indicated that GF reduced the miR-489 content in the HEK-293 cell model and Cur could prevent this downregulation. Then, we planned to explore the character of miR-489 in Cur-triggered attenuation of the Warburg effect under GF condition. Our findings presented that Cur prevented GF-triggered aerobic glycolysis by upregulating miR-489 in HEK-293 cells. Next, we choose the miR-489/LDHA axis for further investigation. We confirmed that Cur prevented GF-triggered aerobic glycolysis via the miR-489/LDHA axis in HEK-293 cells. In conclusion, this study presented that Cur prevented GF-triggered renal injury by restraining aerobic glycolysis via the miR-489/LDHA axis in the HEK-293 cell model.

scholarly journals Involvement of Mitogen-Activated Protein Kinase in Agonist-Induced Phosphorylation of the μ-Opioid Receptor in HEK 293 Cells

2001 ◽  
Vol 74 (1) ◽  
pp. 414-422 ◽  
Author(s):  
Harald Schmidt ◽  
Stefan Schulz ◽  
Marcus Klutzny ◽  
Thomas Koch ◽  
Manuela Händel ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Opioid Receptor ◽  
Mitogen Activated Protein Kinase ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Mitogen Activated Protein ◽  
Μ Opioid Receptor

New C(2)-sulfonyl ester-type Salvinorin A ligands to the kappa-opioid receptor

Planta Medica ◽  
2015 ◽  
Vol 81 (05) ◽  
Author(s):  
PR Polepally ◽  
A Keasling ◽  
K White ◽  
E Vardy ◽  
BL Roth ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Kappa Opioid Receptor ◽  
Salvinorin A ◽  
Kappa Opioid

scholarly journals The Role of Dynorphin and the Kappa Opioid Receptor in Schizophrenia and Major Depressive Disorder: A Translational Approach

2020 ◽  
Author(s):  
Samuel David Clark
Keyword(s):  
Major Depressive Disorder ◽  
Depressive Disorder ◽  
Healthy Volunteers ◽  
Opioid Receptor ◽  
Clinical Data ◽  
Potential Role ◽  
Kappa Opioid Receptor ◽  
Kappa Opioid ◽  
Major Depressive

AbstractThe kappa opioid receptor (KOR) and its endogenous ligands dynorphins (DYN) have been implicated in the development or symptomatology of a variety of neuropsychiatric disorders. This review covers a brief history of the development of KOR agonists and antagonists, their effects in healthy volunteers, and the potential role of DYN/KOR dysfunction in schizophrenia and major depressive disorder from a translational perspective. The potential role of DYN/KOR dysfunction in schizophrenia is based on several lines of evidence. Selective KOR agonists induce affective states in healthy volunteers with similarities to the symptoms of schizophrenia. Studies have shown increased DYN in patients with schizophrenia, although the data have been mixed. Finally, meta-analytic data have shown that opioid antagonists are associated with reductions in the symptoms of schizophrenia. The potential role of DYN/KOR dysfunction in major depressive disorder is also based on a combination of preclinical and clinical data. Selective KOR agonists have shown pro-depressive effects in human volunteers, while selective KOR antagonists have shown robust efficacy in several preclinical models of antidepressant activity. Small studies have shown that nonselective KOR antagonists may have efficacy in treatment-resistant depression. Additionally, recent clinical data have shown that the KOR may be an effective target for treating anhedonia, a finding relevant to both schizophrenia and depression. Finally, recommendations are provided for translating preclinical models for schizophrenia and major depressive disorder into the clinic.

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