Mironov, Sergej L.

[["dc.bibliographiccitation.artnumber","29"],["dc.bibliographiccitation.journal","BMC Neuroscience"],["dc.bibliographiccitation.volume","10"],["dc.contributor.author","Mironov, Sergej L."],["dc.contributor.author","Skorova, Ekaterina Y."],["dc.contributor.author","Taschenberger, Grit"],["dc.contributor.author","Hartelt, N."],["dc.contributor.author","Nikolaev, V. O."],["dc.contributor.author","Lohse, Martin J."],["dc.contributor.author","Kuegler, Sebastian"],["dc.date.accessioned","2018-11-07T08:31:34Z"],["dc.date.available","2018-11-07T08:31:34Z"],["dc.date.issued","2009"],["dc.description.abstract","Background: cAMP is an ubiquitous second messenger mediating various neuronal functions, often as a consequence of increased intracellular Ca(2+) levels. While imaging of calcium is commonly used in neuroscience applications, probing for cAMP levels has not yet been performed in living vertebrate neuronal tissue before. Results: Using a strictly neuron-restricted promoter we virally transduced neurons in the organotypic brainstem slices which contained pre-Botzinger complex, constituting the rhythm-generating part of the respiratory network. Fluorescent cAMP sensor Epac1-camps was expressed both in neuronal cell bodies and neurites, allowing us to measure intracellular distribution of cAMP, its absolute levels and time-dependent changes in response to physiological stimuli. We recorded [cAMP](i) changes in the micromolar range after modulation of adenylate cyclase, inhibition of phosphodiesterase and activation of G-protein-coupled metabotropic receptors. [cAMP](i) levels increased after membrane depolarisation and release of Ca(2+) from internal stores. The effects developed slowly and reached their maximum after transient [Ca(2+)](i) elevations subsided. Ca(2+)-dependent [cAMP](i) transients were suppressed after blockade of adenylate cyclase with 0.1 mM adenylate cyclase inhibitor 2'5'-dideoxyadenosine and potentiated after inhibiting phosphodiesterase with isobutylmethylxanthine and rolipram. During paired stimulations, the second depolarisation and Ca(2+) release evoked bigger cAMP responses. These effects were abolished after inhibition of protein kinase A with H-89 pointing to the important role of phosphorylation of calcium channels in the potentiation of [cAMP](i) transients. Conclusion: We constructed and characterized a neuron-specific cAMP probe based on Epac1-camps. Using viral gene transfer we showed its efficient expression in organotypic brainstem preparations. Strong fluorescence, resistance to photobleaching and possibility of direct estimation of [cAMP] levels using dual wavelength measurements make the probe useful in studies of neurons and the mechanisms of their plasticity. Epac1-camps was applied to examine the crosstalk between Ca(2+) and cAMP signalling and revealed a synergism of actions of these two second messengers."],["dc.identifier.doi","10.1186/1471-2202-10-29"],["dc.identifier.isi","000265852000001"],["dc.identifier.pmid","19327133"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/5797"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/17151"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.relation.issn","1471-2202"],["dc.rights","Goescholar"],["dc.rights.uri","https://goescholar.uni-goettingen.de/licenses"],["dc.title","Imaging cytoplasmic cAMP in mouse brainstem neurons"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]