[HTML][HTML] Mechanisms restricting diffusion of intracellular cAMP

SR Agarwal, CE Clancy, RD Harvey - Scientific reports, 2016 - nature.com
Scientific reports, 2016nature.com
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 …
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.
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