Unit

UNIT 7.12 Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates

  1. Fabian Gusovsky

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0712s05

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Gusovsky, F. 2001. Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates. Current Protocols in Neuroscience. 5:7.12:7.12.1–7.12.11.

Author Information

  1. Eisai Research Institute, Andover, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: NOV 1998

Abstract

cAMP acts as an intracellular mediator of hormone action and the importance of accurate quantitative determination of cAMP levels in cells and tissues is widely recognized. The most utilized procedures for the determination of adenylate cyclase activity in membranes are described here for measuring the conversion of [alpha-32P]ATP into [32P]cAMP after a two-step chromatographic separation. Also critical in signal tranduction is phosphoinositide turnover, which is linked to receptor activation resulting from changes in cytosolic calcium concentrations. Phosphoinositide turnover can be measured as described in this unit by labeling phospholipid pools with [3H]-inositol and then analyzing for tritiated inositol phosphates.