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Imaging calcium in hippocampal presynaptic terminals with a ratiometric calcium sensor in a novel transgenic mouse

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posted on 25.07.2019, 08:50 by I Al-Osta, M Mucha, D Pereda, M Piqué-Gili, AE Okorocha, R Thomas, NA Hartell
Genetically encoded calcium indicators (GECIs) have gained widespread use for measurement of neuronal activity but their low expression levels in transgenic mice tend to limit sensitivity. We have developed a transgenic mouse line (SyG37) that expresses a ratiometric calcium sensor, SyGCaMP2-mCherry, that is expressed throughout the brain but targeted to presynaptic terminals. Within the CA1 and CA3 regions of hippocampus of male and female mice, SyGaMP2 fluorescence responds linearly up to 10 electrical stimuli at frequencies up to 100 Hz and it can detect responses to a single stimulus. Responses in single boutons can be measured using multiphoton microscopy. The ensemble amplitude of SyGCaMP2 responses is a function of the number of stimuli applied and the number of contributing boutons. The peak responses and initial rates of calcium influx in single boutons in CA1 and CA3 were similar but the rate of calcium clearance from CA3 boutons after stimulation was significantly faster. In CA1, DNQX reduced SyGCaMP2 responses to Schaffer collateral stimulation to 86% of baseline indicating that 14% of the total response originated from presynaptic terminals of neurones synaptically driven via AMPA receptors. Theta burst stimulation induced long-term potentiation (LTP) of both SyGCaMP2 and fEPSP responses in both young and 18-month-old mice. The proportion of postsynaptically connected terminals increased significantly to 76% of the total after LTP induction. The SyG37 mouse allows stable optical detection of synaptic activation and connectivity at the single bouton level and can be used to characterize the contributions of presynaptic calcium to synaptic transmission and plasticity.


This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grants BB/K008382/1 and BB/L00691X/1 to NH.



Frontiers in Cellular Neuroscience, 2018, 12:209

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Corrigendum: Imaging Calcium in Hippocampal Presynaptic Terminals With a Ratiometric Calcium Sensor in a Novel Transgenic Mouse Front. Cell. Neurosci. 13:194. doi: 10.3389/fncel.2019.00194 http://hdl.handle.net/2381/45622 In the original article, there was a mistake in Figure 8 as published. During the revision process, panels B-G from Figure 2 were incorrectly duplicated into panels B-G of Figure 8. The data for these figures were correct in the original submission. The corrected Figure 8 appears below. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.