title: Depression of type I diacylglycerol kinases in pancreatic β-cells from male mice results in impaired insulin secretion.
abstract: Diacylglycerol kinase (DGK) catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid. This study investigated the expression and function of DGK in pancreatic β-cells. mRNA expression of type I DGK isoforms (α, β, γ) was detected in mouse pancreatic islets and the β-cell line MIN6. Protein expression of DGKα and DGKγ was also detected in mouse β-cells and MIN6 cells. The type I DGK inhibitor R59949 inhibited high K(+)- and glucose-induced insulin secretion in MIN6 cells. Moreover, single knockdown of DGKα or DGKγ by small interfering RNA slightly but significantly decreased glucose- and high K(+)-induced insulin secretions, and the double knockdown further decreased them to the levels comparable with those induced by R59949. R59949 and DiC8, a membrane permeable DAG analog, decreased intracellular Ca(2+) concentration elevated by glucose and high K(+) in MIN6 cells. Real-time imaging in MIN6 cells expressing green fluorescent protein-tagged DGKα or DGKγ showed that the DGK activator phorbol 12-myristate 13-acetate rapidly induced translocation of DGKγ to the plasma membrane, whereas high K(+) slowly translocated DGKα and DGKγ to the plasma membrane. R59949 increased the DAG content in MIN6 cells when stimulated with high KCl, whereas it did not increase the DAG content but decreased the phosphatidic acid content when stimulated with high glucose. Finally, R59949 was confirmed to inhibit high K(+)-induced insulin secretion from mouse islets and glucose-induced insulin secretion from rat islets. These results suggest that DGKα and DGKγ are present in β-cells and that the depression of these DGKs causes a decrease in intracellular Ca(2+) concentration, thereby reducing insulin secretion.
authors.affiliation: PhD, 52-1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422-8526, Japan. ishikat@u-shizuoka-ken.ac.jp.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kurohane Kaneko
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Keisuke
authors.initials: K
authors.lastname: Motoki
authors.forename: Kunihito
authors.initials: K
authors.lastname: Nakata
authors.forename: Shoko
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Mao
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Daiki
authors.initials: D
authors.lastname: Hayashi
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Shirai
authors.forename: Fumio
authors.initials: F
authors.lastname: Sakane
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Ishikawa
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D019852:Diacylglycerol Kinase / Q000235:genetics / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D010880:Piperidines / Q000494:pharmacology; D020033:Protein Isoforms; D052999:Quinazolinones / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
chemical_list: D007328:Insulin; D010880:Piperidines; D020033:Protein Isoforms; D052999:Quinazolinones; C058544:R 59949; D019852:Diacylglycerol Kinase; D002118:Calcium