Measuring phospholipase D activity in insulin-secreting pancreatic beta-cells and insulin-responsive muscle cells and adipocytes.
Phospholipase D (PLD) is an enzyme producing phosphatidic acid and choline through hydrolysis of phosphatidylcholine. The enzyme has been identified as a member of a variety of signal transduction cascades and as a key regulator of numerous intracellular vesicle trafficking processes. A role for PLD in regulating glucose homeostasis is emerging as the enzyme has recently been identified in events regulating exocytosis of insulin from pancreatic beta-cells and also in insulin-stimulated glucose uptake through controlling GLUT4 vesicle exocytosis in muscle and adipose tissue. We present methodologies for assessing cellular PLD activity in secretagogue-stimulated insulin-secreting pancreatic beta-cells and also insulin-stimulated adipocyte and muscle cells, two of the principal insulin-responsive cell types controlling blood glucose levels.
Cazzolli
Rosanna
R
Department of Medicine, Garvan Institute of Medical Research, Sydney, New South Wales, Australia.
Huang
Ping
P
Teng
Shuzhi
S
Hughes
William E
WE
D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D006801:Humans; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000201:enzymology* / Q000378:metabolism*; D051379:Mice; D032342:Muscle Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects; D010739:Phospholipase D / Q000032:analysis* / Q000378:metabolism*; D010743:Phospholipids / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D013194:Staining and Labeling
D007328:Insulin; D010743:Phospholipids; D010739:Phospholipase D