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  • 1
    Online-Ressource
    Online-Ressource
    Springer Science and Business Media LLC ; 2023
    In:  Prostate Cancer and Prostatic Diseases Vol. 26, No. 2 ( 2023-06), p. 293-301
    In: Prostate Cancer and Prostatic Diseases, Springer Science and Business Media LLC, Vol. 26, No. 2 ( 2023-06), p. 293-301
    Materialart: Online-Ressource
    ISSN: 1365-7852 , 1476-5608
    Sprache: Englisch
    Verlag: Springer Science and Business Media LLC
    Publikationsdatum: 2023
    ZDB Id: 2008886-3
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 2
    Online-Ressource
    Online-Ressource
    Elsevier BV ; 2020
    In:  Bone Reports Vol. 13 ( 2020-10), p. 100443-
    In: Bone Reports, Elsevier BV, Vol. 13 ( 2020-10), p. 100443-
    Materialart: Online-Ressource
    ISSN: 2352-1872
    Sprache: Englisch
    Verlag: Elsevier BV
    Publikationsdatum: 2020
    ZDB Id: 2821774-3
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 3
    Online-Ressource
    Online-Ressource
    The Endocrine Society ; 2023
    In:  Journal of the Endocrine Society Vol. 7, No. Supplement_1 ( 2023-10-05)
    In: Journal of the Endocrine Society, The Endocrine Society, Vol. 7, No. Supplement_1 ( 2023-10-05)
    Kurzfassung: Disclosure: K. Prins: None. M. Huisman: None. R. Mies: None. P.J. Delhanty: None. J.A. Visser: None. Ghrelin is a gut hormone that enhances food intake and growth hormone secretion through its receptor, the G-protein coupled receptor (GPCR) GHSR1a. Recently, we showed that ghrelin interacts with syndecans (SDCs), a family of four membrane proteins that have been linked to obesity in genome-wide association studies. Here, we investigated whether SDCs impact ghrelin signaling at GHSR1a. We used HEK293 cells that expressed GHSR1a and SDC1, SDC2, SDC3, or SDC4 through transient transfection. GHSR1a mainly signals through the Gαq pathway, so we first explored the effects of SDCs on ghrelin-induced intracellular Ca2+ influx. Using a mitochondrial-targeted aequorin, this Ca2+ response can be measured seconds after ghrelin stimulation. We found that all SDCs increased the maximum Ca2+ response at least 4.2-fold (P & lt;0.001), without affecting the EC50. This enhanced Ca2+ response was not caused by increased receptor availability: SDCs reduced cell membrane receptor expression by 51-85% (P & lt;0.001). This low GHSR1a membrane availability contrasted with the increased Ca2+ response, and raised the question whether SDCs could be an alternate receptor for ghrelin. To test this, we transfected cells with SDC1 alone, but this did not permit a ghrelin-stimulated Ca2+ response, indicating that the ghrelin-induced Ca2+ response was GHSR1a-dependent. Moreover, the SDC-potentiated response was also Gαq-dependent, since GNAQ knockout cells transfected with GHSR1a and SDC1 expression plasmids failed to generate a Ca2+ response. However, using a Gq biosensor, we found that the level of activation of Gαq by ghrelin was not affected by SDC1 transfection, suggesting an effect of SDC1 either downstream of Gαq or on the quenching of GHSR. Thus, we investigated the interaction between GHSR1a and β-arrestin 2. Β-arrestin 2 downregulates GHSR function following activation by sterically hindering G-protein coupling and mediating receptor internalization. Four minutes after ghrelin stimulation, SDCs reduced β-arrestin 2 recruitment by 69-87% (P & lt;0.001). SDC-transfected cells also reached a maximum β-arrestin 2 response 8-14 minutes later than GHSR1a only-transfected cells. Notably, these differences in β-arrestin 2 recruitment occurred minutes after the Ca2+ response. These findings suggest that the reduction in β-arrestin 2 recruitment does not cause the increased Ca2+ response in the presence of SDCs, and that the effect of SDCs is probably downstream of Gαq. Interestingly, the effects of SDCs mimic those of melanocortin receptor accessory protein 2 (MRAP2). MRAP2 also enhances the ghrelin-stimulated activation of the Gαq signaling pathway, independently of β-arrestin recruitment. Altogether, we showed that SDCs enhanced Ca2+ influx and reduced β-arrestin 2 recruitment to GHSR1a in response to ghrelin. This could be a novel mechanism through which SDCs affect metabolism and obesity. Presentation: Thursday, June 15, 2023
    Materialart: Online-Ressource
    ISSN: 2472-1972
    Sprache: Englisch
    Verlag: The Endocrine Society
    Publikationsdatum: 2023
    ZDB Id: 2881023-5
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 4
    Online-Ressource
    Online-Ressource
    The Endocrine Society ; 2020
    In:  Journal of the Endocrine Society Vol. 4, No. Supplement_1 ( 2020-05-08)
    In: Journal of the Endocrine Society, The Endocrine Society, Vol. 4, No. Supplement_1 ( 2020-05-08)
    Kurzfassung: Ghrelin, a 28-amino acid peptide gut hormone, occurs in acylated (AG) and unacylated (UAG) variants. AG is a GH secretagogue as well as being orexigenic and diabetogenic, acting via the growth hormone secretagogue receptor (GHSR1a) in the hypothalamus and pituitary. UAG counteracts these metabolic effects through unknown mechanisms. While screening for potential UAG receptor(s) we discovered previously uncharacterised interactions of AG and UAG with five cell membrane proteins (MPs), three of which are known to modulate metabolism. Here, we studied if two of these MPs (MP1 & 2) could modulate GHSR1a signalling by expressing their transgenes in HEK293 cells. As GHSR1a is coupled with calcium signalling via Gq proteins, aequorin luminescence was used to evaluate Ca2+ influx into the cells. Transfected cells were treated with either AG, UAG, or soluble parts of the MPs, or combinations thereof. MP2 markedly enhanced the efficacy (~5.5-fold), but not the potency, of AG-induced Ca2+ influx, whereas MP1 had no effect on Ca2+ influx. Neither MP1 nor MP2 overexpression altered cellular GHSR1a levels. In the absence of GHSR1a, MP2 was unable to stimulate an AG-induced Ca2+ influx. UAG treatment (100nM) had no effect on GHSR1a-mediated Ca2+ influx in the presence or absence of MP2. MP2 is post-translationally modified and we suspected this to be important for its activity. However, removal or blockade of these modifications had no effect on the ability of MP2 to enhance GHSR1a signalling. Moreover, incubating the cells with soluble ectodomain of MP2 did not alter its effect on GHSR1a signalling. Nevertheless, induction of ectodomain shedding with PMA (0.4-1µM) dose-dependently reduced the AG-induced Ca2+ response to 0.5-0.2 of control levels (DMSO) in MP2-GHSR1a co-transfected cells. Unlike MP2, which has a transmembrane and intracellular domain, MP1 is attached to the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor and lacks an intracellular domain. Since MP1 is otherwise structurally similar to MP2, we suspected that the intracellular domain of MP2 may be important for its function. Therefore, we expressed chimeras of MP2 and MP1 in which the GPI linkage site and the transmembrane/intracellular domains were exchanged. The MP2 ectodomain with a GPI-anchor had similar stimulatory effects on GHSR1a signalling as full-length MP2, whereas the MP1 ectodomain with MP2 transmembrane and intracellular domain only enhanced GHSR1a signalling by approximately 3-fold. In conclusion, we have identified a membrane protein as a novel component of the ghrelin signalling pathway that markedly enhances the response of the ghrelin receptor to AG. Our current data suggest its ectodomain is important in mediating this effect. Studies are ongoing to fully delineate the mode of interaction and to determine the role of MP2 in ghrelin signalling in vivo.
    Materialart: Online-Ressource
    ISSN: 2472-1972
    Sprache: Englisch
    Verlag: The Endocrine Society
    Publikationsdatum: 2020
    ZDB Id: 2881023-5
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 5
    Online-Ressource
    Online-Ressource
    Bioscientifica ; 2021
    In:  Journal of Molecular Endocrinology Vol. 66, No. 2 ( 2021-02), p. 83-96
    In: Journal of Molecular Endocrinology, Bioscientifica, Vol. 66, No. 2 ( 2021-02), p. 83-96
    Kurzfassung: Acylated ghrelin (AG) is a gut-derived peptide with growth hormone secretagogue (GHS), orexigenic and other physiological activities mediated by GHS receptor-1a (GHSR). Ghrelin occurs in unacylated form (UAG) with activities opposing AG, although its mechanism of action is unknown. UAG does not antagonize AG at GHSR, and has biological effects on cells that lack this receptor. Because UAG binds to cells, it has been hypothesized that UAG acts via a cell-surface receptor, although this has not been confirmed. This study aimed to identify cell surface proteins to which UAG binds that could modulate or mediate its biological effects. The MCF7 cell-line was used as a model because UAG induces ERK signaling in these cells in the absence of GHSR. Using ligand–receptor capture and LC-MS/MS we identified specific heparan-sulfate proteoglycans (HSPGs) to which UAG interacts on cell surfaces. In line with this, UAG, as well as AG, bind with high affinity to heparin, and heparin and heparinase treatment suppress, whereas HSPG overexpression increases, UAG binding to MCF7 cell surfaces. Moreover, heparin suppresses the ERK response to UAG. However, conversion of the lysines in UAG to alanine, which prevents its binding to heparin and cell surface HSPGs, does not prevent its activation of ERK. Our data show that the interaction of UAG with HSPGs modulates its biological activity in cells. More broadly, the interaction of UAG and AG with HSPGs could be important for the specificity and potency of their biological action in vivo .
    Materialart: Online-Ressource
    ISSN: 0952-5041 , 1479-6813
    Sprache: Unbekannt
    Verlag: Bioscientifica
    Publikationsdatum: 2021
    ZDB Id: 1478171-2
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 6
    Online-Ressource
    Online-Ressource
    The Endocrine Society ; 2021
    In:  Journal of the Endocrine Society Vol. 5, No. Supplement_1 ( 2021-05-03), p. A56-A56
    In: Journal of the Endocrine Society, The Endocrine Society, Vol. 5, No. Supplement_1 ( 2021-05-03), p. A56-A56
    Kurzfassung: Ghrelin, a peptide hormone secreted by the stomach, stimulates both appetite and reward signalling. Its deletion in mice results in poor recovery from metabolic challenges, like starvation, but does not affect food intake or body weight. While sex differences in appetite and feeding behavior have been reported, little is known about the role of ghrelin herein. To investigate this, we used a metabolic cage system to continuously monitor responses of ghrelin-deficient (GKO) and wildtype (WT) mice to three different diets. Male and female mice (5 weeks old) were housed individually in a Promethion system (Sable Systems, USA) and provided one of three diets for 9 weeks: RA, continuous chow with restricted access to a Western-style diet (WD; 2h access, 3d/week) in the light phase; CA, continuous access to both diets; CC, continuous chow. Glucose tolerance was assessed at week 7 by IPGTT; food intake (kcal/g bodyweight), energy expenditure and locomotor activity at week 8; body weight and body composition (EchoMRI, USA) at week 9. On access days, RA mice ate up to 60% of their 24h intake during the WD access period. Following WD access GKO RA mice ate less chow than WT RA mice. Intriguingly, this compensatory reduction in food intake by GKO mice occurred at different times for males and females. GKO RA males ate 45% less chow in the dark phase immediately after WD access (p & lt; 0.001). In contrast, this reduction in food intake (30% less) did not occur until the following, non-access, day in GKO RA females (genotype-sex: p & lt; 0.05). Depending on diet, GKO mice showed differential regulation of energy expenditure in the light phase. Energy expenditure was 6–17% higher in GKO than WT mice in the RA group on access days and in the CA group. On non-access days, however, GKO mice in the RA group expended 13% less energy than WT RA mice (p & lt; 0.005). Regardless of diet, locomotor activity in females was greater than in males (p & lt; 0.001). However, GKO females in the RA and CC groups showed a marked 30% reduction in locomotor activity compared to WTs (genotype-sex: p & lt; 0.05). After nine weeks, neither sex nor genotype effects were seen in body weight gain and composition of RA animals. CA females gained 17% more body weight and had a 6.1% higher fat percentage than CA males (both p & lt; 0.001). In the CC group body weight gain did not differ, but GKO females had 3.1% more fat than WT females (genotype-sex: p & lt; 0.01). Glucose tolerance (AUC) was similar in all groups. In conclusion, we demonstrated that ghrelin deficiency changes the response to the three diets in a sex-dependent manner. Especially, restricted access to WD differentially affected food intake timing and locomotor activity of male and female GKO mice. These results add to the growing body of evidence that ghrelin signalling is sexually dimorphic.
    Materialart: Online-Ressource
    ISSN: 2472-1972
    Sprache: Englisch
    Verlag: The Endocrine Society
    Publikationsdatum: 2021
    ZDB Id: 2881023-5
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 7
    In: Endocrine Abstracts, Bioscientifica, ( 2022-05-07)
    Materialart: Online-Ressource
    ISSN: 1479-6848
    Sprache: Unbekannt
    Verlag: Bioscientifica
    Publikationsdatum: 2022
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 8
    Online-Ressource
    Online-Ressource
    American Physiological Society ; 2022
    In:  American Journal of Physiology-Endocrinology and Metabolism Vol. 322, No. 6 ( 2022-06-01), p. E494-E507
    In: American Journal of Physiology-Endocrinology and Metabolism, American Physiological Society, Vol. 322, No. 6 ( 2022-06-01), p. E494-E507
    Kurzfassung: Binge-eating disorder is the most prevalent eating disorder diagnosed, affecting three times more women than men. Ghrelin stimulates appetite and reward signaling, and loss of its receptor reduces binge-eating behavior in male mice. Here, we examined the influence of ghrelin itself on binge-eating behavior in both male and female mice. Five-wk-old wild-type (WT) and ghrelin-deficient ( Ghrl −/− ) mice were housed individually in indirect calorimetry cages for 9 wks. Binge-like eating was induced by giving mice ad libitum chow, but time-restricted access to a Western-style diet (WD; 2 h access, 3 days/wk) in the light phase (BE); control groups received ad libitum chow (CO), or ad libitum access to both diets (CW). All groups of BE mice showed binge-eating behavior, eating up to 60% of their 24-h intake during the WD access period. Subsequent dark phase chow intake was decreased in Ghrl −/− mice and remained decreased in Ghrl −/− females on nonbinge days. Also, nonbinge day locomotor activity was lower in Ghrl −/− than in WT BE females. Upon euthanasia, Ghrl −/− BE mice weighed less and had a lower lean body mass percentage than WT BE mice. In BE and CW groups, ghrelin and sex altered the expression of genes involved in lipid processing, thermogenesis, and aging in white adipose tissue and livers. We conclude that, although ghrelin deficiency does not hamper the development of binge-like eating, it sex-dependently alters food intake timing, locomotor activity, and metabolism. These results add to the growing body of evidence that ghrelin signaling is sexually dimorphic. NEW & NOTEWORTHY Ghrelin, a peptide hormone secreted from the gut, is involved in hunger and reward signaling, which are altered in binge-eating disorder. Although sex differences have been described in both binge-eating and ghrelin signaling, this interaction has not been fully elucidated. Here, we show that ghrelin deficiency affects the behavior and metabolism of mice in a binge-like eating paradigm, and that the sex of the mice impacts the magnitude and direction of these effects.
    Materialart: Online-Ressource
    ISSN: 0193-1849 , 1522-1555
    Sprache: Englisch
    Verlag: American Physiological Society
    Publikationsdatum: 2022
    ZDB Id: 1477331-4
    SSG: 12
    Standort Signatur Einschränkungen Verfügbarkeit
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