Glucose-mediated control of ghrelin release from primary cultures of gastric mucosal cells

Division of Hypothalamic Research, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390-9077, USA.
AJP Endocrinology and Metabolism (Impact Factor: 3.79). 03/2012; 302(10):E1300-10. DOI: 10.1152/ajpendo.00041.2012
Source: PubMed


The peptide hormone ghrelin is released from a distinct group of gastrointestinal cells in response to caloric restriction, whereas its levels fall after eating. The mechanisms by which ghrelin secretion is regulated remain largely unknown. Here, we have used primary cultures of mouse gastric mucosal cells to investigate ghrelin secretion, with an emphasis on the role of glucose. Ghrelin secretion from these cells upon exposure to different d-glucose concentrations, the glucose antimetabolite 2-deoxy-d-glucose, and other potential secretagogues was assessed. The expression profile of proteins involved in glucose transport, metabolism, and utilization within highly enriched pools of mouse ghrelin cells and within cultured ghrelinoma cells was also determined. Ghrelin release negatively correlated with d-glucose concentration. Insulin blocked ghrelin release, but only in a low d-glucose environment. 2-Deoxy-d-glucose prevented the inhibitory effect of high d-glucose exposure on ghrelin release. mRNAs encoding several facilitative glucose transporters, hexokinases, the ATP-sensitive potassium channel subunit Kir6.2, and sulfonylurea type 1 receptor were expressed highly within ghrelin cells, although neither tolbutamide nor diazoxide exerted direct effects on ghrelin secretion. These findings suggest that direct exposure of ghrelin cells to low ambient d-glucose stimulates ghrelin release, whereas high d-glucose and glucose metabolism within ghrelin cells block ghrelin release. Also, low d-glucose sensitizes ghrelin cells to insulin. Various glucose transporters, channels, and enzymes that mediate glucose responsiveness in other cell types may contribute to the ghrelin cell machinery involved in regulating ghrelin secretion under these different glucose environments, although their exact roles in ghrelin release remain uncertain.

Download full-text


Available from: Ichiro Sakata, Jan 25, 2016
  • Source
    • "Furthermore, the glucoprivic agent 2-deoxy-D-glucose prevents the inhibitory effect of high glucose exposure on ghrelin release, suggesting a requirement for glucose entry into the ghrelin cell and subsequent metabolism for its inhibitory effects on ghrelin secretion [71]. This is further supported by the expression by ghrelin cells of mRNAs encoding several channels and enzymes responsible for mediating the effects glucose responsiveness and metabolism in other cell types, including several facilitative glucose transporters (GLUT1, GLUT4, and GLUT5), hexokinases (including glucokinase), and both components of the pancreatic b-cell KATP channel (the ATP-sensitive potassium channel subunit Kir6.2 and the sulfonylurea type 1 receptor SUR1) [71]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The current study examined potential mechanisms for altered circulating ghrelin levels observed in diet-induced obesity (DIO) and following weight loss resulting from Roux-en-Y gastric bypass (RYGB). We hypothesized that circulating ghrelin levels were altered in obesity and after weight loss through changes in ghrelin cell responsiveness to physiological cues. We confirmed lower ghrelin levels in DIO mice and demonstrated elevated ghrelin levels in mice 6 weeks post-RYGB. In both DIO and RYGB settings, these changes in ghrelin levels were associated with altered ghrelin cell responsiveness to two key physiological modulators of ghrelin secretion – glucose and norepinephrine. In DIO mice, increases in ghrelin cell density within both the stomach and duodenum and in somatostatin-immunoreactive D cell density in the duodenum were observed. Our findings provide new insights into the regulation of ghrelin secretion and its relation to circulating ghrelin within the contexts of obesity and weight loss.
    Full-text · Article · Oct 2014 · Molecular Metabolism
  • Source
    • "2.8. Primary gastric mucosal cell cultures The method was modified from [11] [49] to obtain fragments of gastric glands instead of single cells from C57BL/6 mice [Charles River, Sulzfeld, DE]. Following cervical dislocation, the stomach was tied off at the esophagus and duodenum, excised, turned inside out through an incision in the nonglandular forestomach, rinsed with PBS, inflated with DMEM-F12 and placed in a 50 ml tube containing 10 ml DMEM-F12 with 20 mM HEPES, 1 mg/ml collagenase (Type XI – Sigma Aldrich), and 0.15% BSA (Sigma Aldrich). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The molecular mechanisms regulating secretion of the orexigenic-glucoregulatory hormone ghrelin remain unclear. Based on qPCR analysis of FACS-purified gastric ghrelin cells, highly expressed and enriched 7TM receptors were comprehensively identified and functionally characterized using in vitro, ex vivo and in vivo methods. Five Gαs-coupled receptors efficiently stimulated ghrelin secretion: as expected the β1-adrenergic, the GIP and the secretin receptors but surprisingly also the composite receptor for the sensory neuropeptide CGRP and the melanocortin 4 receptor. A number of Gαi/o-coupled receptors inhibited ghrelin secretion including somatostatin receptors SSTR1, SSTR2 and SSTR3 and unexpectedly the highly enriched lactate receptor, GPR81. Three other metabolite receptors known to be both Gαi/o- and Gαq/11-coupled all inhibited ghrelin secretion through a pertussis toxin-sensitive Gαi/o pathway: FFAR2 (short chain fatty acid receptor; GPR43), FFAR4 (long chain fatty acid receptor; GPR120) and CasR (calcium sensing receptor). In addition to the common Gα subunits three non-common Gαi/o subunits were highly enriched in ghrelin cells: GαoA, GαoB and Gαz. Inhibition of Gαi/o signaling via ghrelin cell-selective pertussis toxin expression markedly enhanced circulating ghrelin. These 7TM receptors and associated Gα subunits constitute a major part of the molecular machinery directly mediating neuronal and endocrine stimulation versus metabolite and somatostatin inhibition of ghrelin secretion including a series of novel receptor targets not previously identified on the ghrelin cell.
    Full-text · Article · Nov 2013 · Molecular Metabolism
  • Source
    • "In addition to its potent orexigenic effect, ghrelin signals and modulates glycemic state. For example, insulin-induced hypoglycemia elevates gastric ghrelin mRNA and exposure of cultured gastric ghrelin cells to low ambient glucose levels potentiates ghrelin release [15] [16]. Furthermore , ghrelin administration to rodents increases blood glucose, raises glucagon and growth hormone levels, lowers insulin levels and attenuates insulin responses during glucose tolerance testing [17] [18] [19] [20]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The hormone ghrelin stimulates eating and helps maintain blood glucose upon caloric restriction. While previous studies have demonstrated that hypothalamic arcuate AgRP neurons are targets of ghrelin, the overall relevance of ghrelin signaling within intact AgRP neurons is unclear. Here, we tested the functional significance of ghrelin action on AgRP neurons using a new, tamoxifen-inducible AgRP-CreERT2 transgenic mouse model that allows spatiotemporally-controlled re-expression of physiological levels of ghrelin receptors (GHSRs) specifically in AgRP neurons of adult GHSR-null mice that otherwise lack GHSR expression. AgRP neuron-selective GHSR re-expression partially restored the orexigenic response to administered ghrelin and fully restored the lowered blood glucose levels observed upon caloric restriction. The normalizing glucoregulatory effect of AgRP neuron-selective GHSR expression was linked to glucagon rises and hepatic gluconeogenesis induction. Thus, our data indicate that GHSR-containing AgRP neurons are not solely responsible for ghrelin's orexigenic effects but are sufficient to mediate ghrelin's effects on glycemia.
    Full-text · Article · Oct 2013 · Molecular Metabolism
Show more