A Dual Nanosensor Approach to Determine the Cytosolic Concentration of ATP in Astrocytes

Frontiers in Cellular Neuroscience
Susanne KöhlerUlrike Winkler

Abstract

Adenosine triphosphate (ATP) is the central energy carrier of all cells and knowledge on the dynamics of the concentration of ATP ([ATP]) provides important insights into the energetic state of a cell. Several genetically encoded fluorescent nanosensors for ATP were developed, which allow following the cytosolic [ATP] at high spatial and temporal resolution using fluorescence microscopy. However, to calibrate the fluorescent signal to [ATP] has remained challenging. To estimate basal cytosolic [ATP] ([ATP]0) in astrocytes, we here took advantage of two ATP nanosensors of the ATeam-family (ATeam1.03; ATeam1.03YEMK) with different affinities for ATP. Altering [ATP] by external stimuli resulted in characteristic pairs of signal changes of both nanosensors, which depend on [ATP]0. Using this dual nanosensor strategy and epifluorescence microscopy, [ATP]0 was estimated to be around 1.5 mM in primary cultures of cortical astrocytes from mice. Furthermore, in astrocytes in acutely isolated cortical slices from mice expressing both nanosensors after stereotactic injection of AAV-vectors, 2-photon microscopy revealed [ATP]0 of 0.7 mM to 1.3 mM. Finally, the change in [ATP] induced in the cytosol of cultured cortical astrocytes by applic...Continue Reading

References

Oct 25, 1994·Proceedings of the National Academy of Sciences of the United States of America·L Pellerin, P J Magistretti
Oct 21, 1998·Developmental Neuroscience·R Dringen, B Hamprecht
Apr 11, 2001·Trends in Endocrinology and Metabolism : TEM·M Guzmán, C Blázquez
Mar 22, 2003·The Journal of Biological Chemistry·Marcus FehrWolf B Frommer
Oct 29, 2003·The Neuroscientist : a Review Journal Bringing Neurobiology, Neurology and Psychiatry·Helle S WaagepetersenArne Schousboe
Dec 16, 2004·Journal of Neural Transmission·P J Magistretti, J-Y Chatton
Jun 22, 2006·Journal of Neurochemistry·Lasse K BakHelle S Waagepetersen
Nov 9, 2007·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ditte LovattMaiken Nedergaard
Oct 23, 2008·Brain Cell Biology·S Andrew HiresLoren L Looger
Jan 6, 2009·Nature Methods·Jim BergGary Yellen
Sep 2, 2009·Proceedings of the National Academy of Sciences of the United States of America·Hiromi ImamuraHiroyuki Noji
Nov 3, 2009·Brain Research Reviews·L Felipe Barros, Joachim W Deitmer
Dec 17, 2009·Brain Research Reviews·Vitali Matyash, Helmut Kettenmann
Mar 11, 2010·Nature Reviews. Neuroscience·Klaus-Armin Nave
Jul 27, 2010·Current Opinion in Neurobiology·Ye Zhang, Ben A Barres
Sep 22, 2010·The Biochemical Journal·Clara BermejoWolf B Frommer
Oct 5, 2010·Frontiers in Neuroenergetics·Carla X BittnerL Felipe Barros
Nov 12, 2010·Nature·Klaus-Armin Nave
Mar 25, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Carla X BittnerL Felipe Barros
Apr 14, 2011·Journal of Neuroscience Research·Franziska Wilhelm, Johannes Hirrlinger
Oct 7, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Iván RuminotL Felipe Barros
Feb 1, 2012·Glia·T Sotelo-HitschfeldL F Barros
Feb 4, 2012·Journal of Neurochemistry·Robert P RequardtJohannes Hirrlinger
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Jan 29, 2013·Biochimica Et Biophysica Acta·Vadim Pérez Koldenkova, Takeharu Nagai
Sep 21, 2013·Methods in Molecular Biology·Yin Pun Hung, Gary Yellen
Jan 8, 2014·Molecular and Cellular Neurosciences·J ToloeS L Mironov
Jan 28, 2014·PloS One·Alejandro San MartínL Felipe Barros
Feb 18, 2014·Cell·Vidhya RangarajuTimothy A Ryan
Mar 29, 2014·The Biochemical Journal·Ignacio Fernández-Moncada, L Felipe Barros
Aug 19, 2014·Glia·Johannes Hirrlinger, Klaus-Armin Nave
Nov 22, 2014·Cold Spring Harbor Perspectives in Biology·Omer Ali BayraktarDavid H Rowitch

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fluorescence imaging

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