Spatially compartmentalized phase regulation of a Ca2+-cAMP-PKA oscillatory circuit
Data files
Nov 16, 2020 version files 3.01 MB
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A79-CiCeEpac2Camps_AC8_1-106_allcells.xlsx
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A79-CiCeEpac2Camps_AC8_1-106_filtered.xlsx
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A79-CiCeEpac2Camps_allcells.xlsx
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A79-CiCeEpac2Camps_filtered.xlsx
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AKAP_AKAR_TEA_btenner.xlsx
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btenner_dryad_readme.txt
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lynAKAR_TEA_btenner.xlsx
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lynCiCeEpacCamps__AC8.xlsx
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MIN6_repeats_summary_20180828.xlsx
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Supp2a_lyncice_8mmibmx.xlsx
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Supp2b_lyncice_milrinone.xlsx
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Supp2c_lyncice_roli_Dish1.xlsx
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Supp2c_lyncice_roli_Dish2.xlsx
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Abstract
Signaling networks are spatiotemporally organized in order to sense diverse inputs, process information, and carry out specific cellular tasks. In pancreatic β cells, Ca2+, cyclic adenosine monophosphate (cAMP), and Protein Kinase A (PKA) exist in an oscillatory circuit characterized by a high degree of feedback. Here, we describe a mode of regulation within this circuit involving a spatial dependence of the relative phase between cAMP, PKA, and Ca2+. We show that nanodomain clustering of Ca2+-sensitive adenylyl cyclases drives oscillations of local cAMP levels to be precisely in-phase with Ca2+ oscillations, whereas Ca2+-sensitive phosphodiesterases maintain out-of-phase oscillations outside of the nanodomain. Disruption of this precise phase relationship perturbs Ca2+ oscillations, suggesting the relative phase within an oscillatory circuit can encode specific functional information. This work unveils a novel mechanism of cAMP compartmentation utilized for localized tuning of an oscillatory circuit and has broad implications for the spatiotemporal regulation of signaling networks.
MIN6 pancreatic beta cells expressing FRET-based biosensors were imaging using an epifluorescent microscope. FRET ratio was calculated using sensitized emission channel / donor excited channel (PKA sensor) or vice versa for cAMP (inverse FRET sensor). Single-cell time traces were collected and analyzed with MATLAB to study compartmentalized oscillatory dynamics.
Please read the readme.