Mechanisms Restricting Diffusion of Intracellular cAMP

Shailesh R. Agarwal, Colleen E Clancy, Robert D. Harvey

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Although numerous receptors stimulate cAMP production in a wide array of cells, many elicit distinct, highly localized responses, implying that the subcellular distribution of cAMP is not uniform. One often used explanation is that phosphodiesterases, which breakdown cAMP, act as functional barriers limiting diffusion. However, several studies refute the notion that this is sufficient, suggesting that phosphodiesterase-independent movement of cAMP must occur at rates slower than free diffusion. But, until now this has never been demonstrated. Using Raster Image Correlation Spectroscopy (RICS), we measured the diffusion coefficient of a fluorescently-labeled cAMP derivative (φ450-cAMP) as well as other fluorescent molecules in order to investigate the role that molecular size, cell morphology, and buffering by protein kinase A (PKA) play in restricting cAMP mobility in different cell types. Our results demonstrate that cytosolic movement of cAMP is indeed much slower than the rate of free diffusion and that interactions with PKA, especially type II PKA associated with mitochondria, play a significant role. These findings have important implications with respect to cAMP signaling in all cells.

Original languageEnglish (US)
Article number19577
JournalScientific Reports
Volume6
DOIs
StatePublished - Jan 22 2016

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Phosphoric Diester Hydrolases
Cyclic AMP-Dependent Protein Kinases
Cyclic AMP-Dependent Protein Kinase Type II
Cyclic AMP Receptors
Cell Size
Spectrum Analysis
Mitochondria

ASJC Scopus subject areas

  • General

Cite this

Mechanisms Restricting Diffusion of Intracellular cAMP. / Agarwal, Shailesh R.; Clancy, Colleen E; Harvey, Robert D.

In: Scientific Reports, Vol. 6, 19577, 22.01.2016.

Research output: Contribution to journalArticle

Agarwal, Shailesh R. ; Clancy, Colleen E ; Harvey, Robert D. / Mechanisms Restricting Diffusion of Intracellular cAMP. In: Scientific Reports. 2016 ; Vol. 6.
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