Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo

Efetova, M., Petereit, L., Rosiewicz, K., Overend, G. , Haussig, F., Hovemann, B.T., Cabrero, P. , Dow, J.A.T. and Schwarzel, M. (2013) Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo. Journal of Cell Science, 126(3), pp. 778-788. (doi:10.1242/jcs.114140)

Efetova, M., Petereit, L., Rosiewicz, K., Overend, G. , Haussig, F., Hovemann, B.T., Cabrero, P. , Dow, J.A.T. and Schwarzel, M. (2013) Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo. Journal of Cell Science, 126(3), pp. 778-788. (doi:10.1242/jcs.114140)

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Abstract

Cyclic AMP (cAMP) is a ubiquitous second messenger that regulates a variety of essential processes in diverse cell types, functioning via cAMP-dependent effectors such as protein kinase A (PKA) and/or exchange proteins directly activated by cAMP (EPAC). In an intact tissue it is difficult to separate the contribution of each cAMP effector in a particular cell type using genetic or pharmacological approaches alone. We, therefore, utilized optogenetics to overcome the difficulties associated with examining a multicellular tissue. The transgenic photoactive adenylyl cyclase bPAC can be activated to rapidly and reversibly generate cAMP pulses in a cell-type-specific manner. This optogenetic approach to cAMP manipulation was validated in vivo using GAL4-driven UAS–bPAC in a simple epithelium, the Drosophila renal (Malpighian) tubules. As bPAC was expressed under the control of cell-type-specific promoters, each cAMP signal could be directed to either the stellate or principal cells, the two major cell types of the Drosophila renal tubule. By combining the bPAC transgene with genetic and pharmacological manipulation of either PKA or EPAC it was possible to investigate the functional impact of PKA and EPAC independently of each other. The results of this investigation suggest that both PKA and EPAC are involved in cAMP sensing, but are engaged in very different downstream physiological functions in each cell type: PKA is necessary for basal secretion in principal cells only, and for stimulated fluid secretion in stellate cells only. By contrast, EPAC is important in stimulated fluid secretion in both cell types. We propose that such optogenetic control of cellular cAMP levels can be applied to other systems, for example the heart or the central nervous system, to investigate the physiological impact of cAMP-dependent signaling pathways with unprecedented precision.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Cabrero, Mr Pablo and Overend, Dr Gayle and Dow, Professor Julian
Authors: Efetova, M., Petereit, L., Rosiewicz, K., Overend, G., Haussig, F., Hovemann, B.T., Cabrero, P., Dow, J.A.T., and Schwarzel, M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Journal of Cell Science
Publisher:The Company of Biologists Ltd
ISSN:0021-9533
ISSN (Online):1477-9137
Copyright Holders:Copyright © 2013 The Company of Biologists Ltd.
First Published:First published in Journal of Cell Science 126(3):778-788
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
571431Extraordinary pH - the mechanism of generation of pH 12 in living systemsJulian DowBiotechnology and Biological Sciences Research Council (BBSRC)BB/J002143/1RI MOLECULAR CELL & SYSTEMS BIOLOGY
503671Unravelling the function of the Nha gene familyJulian DowBiotechnology and Biological Sciences Research Council (BBSRC)BB/H001042/1RI MOLECULAR CELL & SYSTEMS BIOLOGY