Using green fluorescent protein to understand the mechanisms of G-protein-coupled receptor regulation

Ferguson, Ssg

doi:10.1590/s0100-879x1998001100016

Cited by 7 publications

(5 citation statements)

References 47 publications

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“…Both the extent and kinetics of agonist‐induced translocation were enhanced by co‐transfection of GRK2. Equivalent results were produced with transient expression of the dopamine D 1A receptor (Barak et al ., 1997) and in review (Ferguson, 1998; Ferguson et al ., 1998) these authors have indicated that the basic features of agonist‐induced β‐arrestin 2‐GFP translocation can be observed for a wide range of GPCRs. One GPCR which is highly resistant to agonist‐induced short‐term desensitization is the gonadotrophin releasing hormone (GnRH) receptor.…”

Section: Introductionmentioning

confidence: 99%

Exploring the dynamics of regulation of G protein‐coupled receptors using green fluorescent protein

Milligan

1999

British J Pharmacology

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Section: Introductionmentioning

confidence: 99%

Exploring the dynamics of regulation of G protein‐coupled receptors using green fluorescent protein

Milligan

1999

British J Pharmacology

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“…GPCRs can be activated by multiple ligands and exhibit the capacity to couple to numerous intracellular signal transduction pathways (18). Oxidative stresses have recently been shown to be generated by agonists of several members of the GPCR superfamily (19).…”

Section: Discussionmentioning

confidence: 99%

Effect of penehyclidine hydrochloride on β-arrestin-1 expression in lipopolysaccharide-induced human pulmonary microvascular endothelial cells

Zhan

Xiao

Zhang

et al. 2013

Braz J Med Biol Res

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β-arrestins are expressed proteins that were first described, and are well-known, as negative regulators of G protein-coupled receptor signaling. Penehyclidine hydrochloride (PHC) is a new anti-cholinergic drug that can inhibit biomembrane lipid peroxidation, and decrease cytokines and oxyradicals. However, to date, no reports on the effects of PHC on β-arrestin-1 in cells have been published. The aim of this study was to investigate the effect of PHC on β-arrestin-1 expression in lipopolysaccharide (LPS)-induced human pulmonary microvascular endothelial cells (HPMEC). Cultured HPMEC were pretreated with PHC, followed by LPS treatment. Muscarinic receptor mRNAs were assayed by real-time quantitative PCR. Cell viability was assayed by the methyl thiazolyl tetrazolium (MTT) conversion test. The dose and time effects of PHC on β-arrestin-1 expression in LPS-induced HPMEC were determined by Western blot analysis. Cell malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were measured. It was found that the M3 receptor was the one most highly expressed, and was activated 5 min after LPS challenge. Furthermore, 2 μg/mL PHC significantly upregulated expression of β-arrestin-1 within 10 to 15 min. Compared with the control group, MDA levels in cells were remarkably increased and SOD activities were significantly decreased in LPS pretreated cells, while PHC markedly decreased MDA levels and increased SOD activities. We conclude that PHC attenuated ROS injury by upregulating β-arrestin-1 expression, thereby implicating a mechanism by which PHC may exert its protective effects against LPS-induced pulmonary microvascular endothelial cell injury.

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“…The sites of phosphorylation have been mapped mainly to the C-terminal tail and have been linked regulatory processes, such as desensitization and internalization [11]. The D-R-Y sequence, positions at 145-147, is highly conserved (Fig.…”

Section: Sequences Analysis Of Mvtrmentioning

confidence: 99%

Characterization and Distribution of an Arginine Vasotocin Receptor in Mouse

Usui

Aoshima

Yamamoto

et al. 2006

The Journal of Veterinary Medical Science

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ABSTRACT. A cDNA, which has a high homology with teleost Platichthys flesus [Arg 8 ] vasotocin (AVT) receptor (GenBank: AK033957), was found in mouse genome database. Analyses of the deduced amino acid sequence revealed that a cDNA has several features of AVT receptor. We tentatively named it as a mouse vasotocin receptor (MVTR [Arg 8 ] vasotocin (AVT) is one of neurohypophyseal hormones [40]. Neurohypophyseal hormones are composed of nine amino acid structures with a disulphide bridge linking two cystines at position 1 and 6 to give a ring structure. AVT possess a hybrid structure, having the C-terminal sequence of [Arg 8 ] vasopressin (AVP) and the N-terminal ring of oxytocin (OT).The role of AVT and its receptor have been well characterized in lower vertebrates including lungfish, amphibians, reptiles and birds [12]. AVT is synthesized in the cell bodies of magnocellular neurons of the hypothalamic pre-optic nucleus as a larger precursor molecule, called pro-vasotocin, and is stored in and released from the pituitary [28]. In the central nervous system, AVT works as a neuromodulator to control reproductive behavior [34,38]. In the periphery, AVT plays a role in regulating osmotic and electrolyte balance and blood pressure [13].A few studies about AVT and its receptor have been reported in mammals. Some reports showed the presence of AVT in the pineal gland of sheep [24], bovine [2] and rat [31,33]. In addition, in vitro experiments also showed that rat pineal releases AVT when it is stimulated by acetylcholine [37]. It has been observed that AVT has several functions in mammals [15]. Most effects of AVT on mammalian brain have been explained by cross-reaction with AVP or OT receptors. However, the effects on sleep-wake cycles such as the cat REM sleep and LH releasing are selective to AVT. In mature cats, injection of AVT into third ventricle decreases the amount of REM sleep, whereas injection of AVT antiserum increases REM sleep. Peripheral administration of AVT in newborn rats increases the quiet state sleep and decreases the active state sleep [17]. Electrophysiological studies on neurons in the rat brain have also shown that the effects of AVT are more potent than either those of AVP and OT [18,26]. From all these data, it has been speculated that there exist AVT-specific receptor in mammals.Recent reports took advantage of the FANTOM2 (Functional Annotation Meeting of Mouse cDNA 2) project, which aimed to collect full-length cDNAs inclusively from mouse tissues, and found 410 candidates for G protein-coupled receptor (GPCR) cDNAs [20]. Forty-eight genes of them were new in mouse and their characters are still not known. In this experiment, we searched for cDNAs encoding AVT receptor in mouse genome database using several sequence analyses. One of the cDNAs was predicted to be a cDNA encoding AVT receptor in mouse. To investigate whether it is a functional AVT receptor, we expressed it in Xenopus oocytes and measured the response against AVT. Distribution of its mRNA was examined by RT-PCR and in situ ...

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Using green fluorescent protein to understand the mechanisms of G-protein-coupled receptor regulation

Cited by 7 publications

References 47 publications

Exploring the dynamics of regulation of G protein‐coupled receptors using green fluorescent protein

Exploring the dynamics of regulation of G protein‐coupled receptors using green fluorescent protein

Effect of penehyclidine hydrochloride on β-arrestin-1 expression in lipopolysaccharide-induced human pulmonary microvascular endothelial cells

Characterization and Distribution of an Arginine Vasotocin Receptor in Mouse

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