Add time:08/22/2019         Source:sciencedirect.com

Production of inositol 1,4,5-trisphosphate (IP3) in cells results in the mobilization of intracellular calcium. Therefore, the dynamics of IP3 metabolism is important for calcium dependent processes in cells. This report investigates the coupling of mAChRs to the inositol lipid pathway in the CNS of the larval Manduca sexta. Stimulation of intact abdominal ganglia prelabeled with [3H]-inositol using a muscarinic agonist, oxotremorine-M (oxo-M), increased total inositol phosphate levels in a dose dependent manner (EC50=4.23 μM). These inositol phosphates consisted primarily of inositol 1,4-bisphosphate (IP2) and inositol monophosphate (IP1). Similarly, when nerve cord homogenates were provided with [3H]-phosphatidylinositol 4,5-bisphosphate ([3H]-PIP2) (10–13 μM) the predominant products were IP2 and IP1. In contrast, incubation of purified membranes with 1 mM oxo-M in the presence of 100 μM GTPγS and [3H]-PIP2 increased IP3 levels, suggesting that the direct activation of phospholipase C (PLC) by mAChRs occurs in a membrane delimited process. Together, these results suggest that in the intact nerve cord and in crude homogenates, a cytosolic 5-phosphatase quickly metabolizes IP3 to produce to IP2 and IP1. This enzyme was kinetically characterized using IP3 (Km=43.7 μM, Vmax=864 pmoles/min/mg) and IP4 (Km=0.93 μM, Vmax=300pmoles/min/mg) as substrates. The enzyme activity can be potently inhibited by two IP thiol compounds; IP3S3 (1,4,6) and IP3S3 (2,3,5), that show complex binding kinetics (Hill numbers<1) and can distinguish different forms of the 5-phosphatase in purified membranes. These two inhibitors could be very useful tools to determine the role of the inositol lipid pathway in neuroexcitability.

We also recommend Trading Suppliers and Manufacturers of inositol 1,4-bisphosphate 5-phosphorothioate (cas 116778-73-5). Pls Click Website Link as below: cas 116778-73-5 suppliers