Spatially Resolved Dynamics of cAMP and Protein Kinase A Subunits in Aplysia Sensory Neurons

Cyclic adenosine monophosphate (cAMP)-dependent protein kinase, labeled with fluorescein and rhodamine on the catalytic and regulatory subunits, respectively, was injected into Aplysia sensory neurons either in culture or in intact cell clusters. Energy transfer between the subunits, a measure of cy...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1993-04, Vol.260 (5105), p.222-226
Main Authors: Bacskai, Brian J., Hochner, Binyamin, Mahaut-Smith, Martyn, Adams, Stephen R., Kaang, Bong-Kiun, Kandel, Eric R., Tsien, Roger Y.
Format: Article
Language:English
Subjects:
1-Methyl-3-isobutylxanthine - pharmacology
Animals
Aplysia
Biochemistry. Physiology. Immunology
Biological and medical sciences
Cell Compartmentation
Cell nucleus
Cell Nucleus - enzymology
Cell Nucleus - metabolism
Cells, Cultured
Concentration camps
Cultured cells
Cyclic AMP - metabolism
Cytoplasm
Cytoplasm - enzymology
Cytoplasm - metabolism
Diffusion
Fluorescein
Fluoresceins
Fluorescence
Fundamental and applied biological sciences. Psychology
Invertebrates
Marine
Microinjections
Mollusca
Neurons
Neurons, Afferent - drug effects
Neurons, Afferent - enzymology
Neurons, Afferent - metabolism
Physiology. Development
Pipettes
Protein Kinases - metabolism
Rhodamines
Sensory neurons
Serotonin - pharmacology
Serotonin receptors
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Summary:Cyclic adenosine monophosphate (cAMP)-dependent protein kinase, labeled with fluorescein and rhodamine on the catalytic and regulatory subunits, respectively, was injected into Aplysia sensory neurons either in culture or in intact cell clusters. Energy transfer between the subunits, a measure of cytosolic cAMP concentration ([cAMP]), and compartmentation of the dissociated subunits were monitored by confocal fluorescence microscopy. Bath application of serotonin produced a much greater elevation of [cAMP] in the processes than in the central bodies of the neurons. The resulting gradients must drive a sizable centripetal flux of cAMP because direct microinjection of cAMP showed that it diffused readily. Perinuclear increases in [cAMP] slowly caused the translocation of the freed catalytic subunit into the nucleus to an extent proportional to the percentage of its dissociation from the regulatory subunit.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.7682336