Biochemical and Biophysical Research Communications
Identification of N-arachidonylglycine, U18666A, and 4-androstene-3,17-dione as novel insulin Secretagogues
Section snippets
Materials and methods
Rat islet cells preparation and culture. Islets were prepared from six male Wistar rats aged 6–7-weeks with free access to food and water. Pancreatic islets were isolated by collagenase treatment followed by Ficoll gradient separation. Briefly, pancreas was inflated by perfusion with 1 mg/ml of collagenase (Wako Pure Chemical Industries, Osaka, Japan) in Hanks’ solution (Sigma) containing 5.6 mM glucose. Pancreas was dissected and then incubated with collagenase at 37 °C for 17 min. After
[Ca2+]i imaging assay system and purification of peptide hormones
To establish a highly sensitive and reproducible physiological screening system for insulinotropic modulators, we employed primary rat pancreatic β-cells and optimized the single-cell calcium imaging technique as described under Materials and methods. This system allowed us to screen 150 samples in a single 5-h experiment, using islets from six animals. This throughput was sufficient to purify insulinotropic peptides from various tissue extracts and screen hundreds of chemical compounds. To
Discussion
We describe here a single-cell Ca2+ imaging assay in primary pancreatic β-cells that is sensitive, reproducible, and efficient enough to permit comprehensive screening of endogenous insulinotropic peptides from various tissue extracts. Our system allows us to screen ∼150 samples in a single 5-h experiment. The technique also allows us to monitor individual islet cells simultaneously and in parallel, which is crucial to avoid misleading false positive signals. However, our system may not permit
Acknowledgments
We thank Dr. Robert W. Rawson and Dr. Patrick C. Reid for helpful discussions, and Yasuyo Urashima, Aoi Uchida, and Satomi Takahashi for excellent technical assistance. This work was supported through Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science and Technology of the Japanese Government and by Exploratory Research for Advanced Technology/Japan Science and Technology Agency (Yanagisawa orphan receptor project). M.Y. is
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2022, Food Science and Human WellnessCitation Excerpt :N-arachidonoyl glycine (NAGly) is a product of an enzymatic synthesis of arachidonic acid and glycine followed by degradation by fatty acid amide hydrolase; the resulting fat amino acid promotes insulin secretion; it has a structural similarity with anandamide (AEA) found mainly in the spinal cord, small intestine, pancreas, and in a variety of animal tissues [48,49]. Ikeda et al. [50] found that in pancreatic beta cells, NAGly increased the [Ca2+]i through voltage-dependent Ca2+ channels (VDCC), resulting in exocytosis of insulin-containing vesicle, thereby promoting the release of insulin to maintain glucose levels in the body. This study was carried out in HFD joint induced by STZ; it is a typical pathological model of “three more and one less,” symptoms, weight loss, and “insulin secretion failure period”; NAGly level was lower in the T2DM group compared to Con (Fig. 6 (18)); following EEP intervention, it rose to a higher level, possibly by stimulating receptors on VDCC, increasing [Ca2+]i, and inhibiting the decomposition of peripheral adipose tissue to improve the body’s glucolipid metabolic disorders.
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2014, Bioorganic and Medicinal ChemistryN-Acyl taurines trigger insulin secretion by increasing calcium flux in pancreatic β-cells
2013, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Expression of TRPV4 has been detected in mouse pancreas and MIN6 cells and may be involved in the increase in intracellular calcium in response to human islet amyloid polypeptide fibril formation [11]. Another N-acyl amino acid, N-arachidonoyl glycine (NAGly), has been identified as a novel insulin secretagogue in primary β-cells [12], although the insulinotropic action of NAGly in rat islet β-cells occurs via voltage-dependent Ca2+ channels (VDCC), rather than via activation mechanism of the TRPV1 channel [12]. The present study was carried out to examine if N-acyl taurines (specifically N-arachidonoyl taurine and N-oleoyl taurine) play a role in insulin secretion in pancreatic β-cells.
A synaptogenic amide N-docosahexaenoylethanolamide promotes hippocampal development
2011, Prostaglandins and Other Lipid MediatorsCitation Excerpt :A number of these N-acylamides have bioactivity. For example, N-arachidonoylglycine inhibits pain [47,48], is an insulin secretagogue [49], a ligand for the orphan receptors GPR18 and GPR92 [50,51], and a reversible inhibitor of the glycine transporter GLYT2a [52]. N-palmitoylglycine increases Ca2+ influx in a dorsal root ganglion-like cell line and is antinociceptive [53], and N-arachidonoylGABA also suppresses pain [47].
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These authors contributed equally to this work.