Decoding neuronal spiking history using a photoconvertible activity indicator
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2018-09-01
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en
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The calcium-modulated photoactivatable ratiometric integrator (CaMPARI) is a recently
developed Ca2+ sensor that achieves time specific neuronal activity marking by the implementation of
Ca2+ dependency together with photo-activatability in its structure. Green fluorescent CaMPARI protein
(baseline of CaMPARI), converts to red fluorescent CaMPARI protein, in order words marks the neuron,
only when user controlled violet light illumination coincides with elevated intracellular Ca2+ levels. Here,
we examined the relationship between the photoconversion rate and the applied violet light and
demonstrated the importance of the light optimization in CaMPARI applications. Furthermore, we
characterized three different variants of CaMPARI with distinct affinities for Ca2+,to our knowledge, for
the first time. CaMPARI indirectly preserves [Ca2+] elevation history of a neuron as the converted red
CaMPARI protein, moreover total amount of red CaMPARI protein is limited by the available green
CaMPARI protein. Therefore, we predicted that activation in vivo would occlude conversion ex vivo.
Finally, we proposed a framework for the use of CaMPARI that can utilize the decryption of the spiking
history.
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Faculteit der Sociale Wetenschappen