Journal: eLife
Article Title: The fate of hippocampal synapses depends on the sequence of plasticity-inducing events
doi: 10.7554/elife.39151
Figure Lengend Snippet: Figure 2. with two supplements: Characterization of oTFS-induced LTP. (A) Changes in excitatory postsynaptic current (EPSC) amplitude in reporter neurons immediately after and 30 min after oTFS in the absence (left) or presence (right) of the NMDA receptor antagonist APV during oTFS. EPSCs were significantly increased after 30 min (p=0.012, n = 20 slice cultures). The increase was blocked by APV (p=0.69, n = 6 slice cultures). (B) Relative change of average excitatory Ca2+ transients (EPCaTs) in individual spines 30 min after the oTFS protocol plotted against the average spine Ca2+ during oTFS. In experiments indicated by filled red circles, APV was present during oTFS. (C) EPSCaT amplitude (p=0.0008, n = 20 slice cultures) and EPSCaT potency (successes only, p=0.0025) but not EPSCaT probability (PCa, p>0.05) were increased 30 min after oTFS in experiments where complex spike bursts (CSBs) were induced during oTFS. (D) Maximum intensity projections of mCerulean fluorescence in dendritic segment harboring a responding spine that was successfully potentiated (blue arrowhead). Volume of oTFS spines (p=0.002, n = 26 spines) and nearest (p=0.0001, n = 45 spines) but not distant neighbors (p=0.83, n = 58 spines) was increased 30 min after oTFS in experiments where CSBs were induced during oTFS. (E) Spine volume was not increased when NMDA receptors were blocked with APV during oTFS (p>0.05, n = 7 spines). Figure 2 continued on next page
Article Snippet: DOI: https://doi.org/10.7554/eLife.39151 12 of 18 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (R. norvegicus, male) Wistar Janvier RjHAN:WI bred in the animal facility, UKE Hamburg Genetic reagent (Clamydomonas reinhardtii) ChR2(ET/TC) doi: 10.1073/ pnas.1017210108 channelr hodopsin Genetic reagent (Aequorea victoria) GCaMP6s doi: 10.1038/ nature12354 calcium indicator Genetic reagent (A. victoria) mCerulean doi: 10.1038/ nbt945 fluorescent protein Transfected construct (R. norvegicus) ChR2(ET/TC) 2Asynaptophysintdimer2 doi: 10.1073/ pnas.1315926110 transfection of CA3 neurons Recombinant DNA reagent rAAV2/7 Vector Facility UKE Hamburg viral vector Chemical compound, drug APV Tocris Bioscience CAS Number 79055-68-8 NMDA receptor blocker Software, algorithm ScanImage3.8 DOI: 10.1186/ 1475-925X-2–13 modified for arbitrary line scans Slice culture preparation and transfection Hippocampal slice cultures from male Wistar rats were prepared at postnatal day 4–5 as described (Gee et al., 2017).
Techniques: Fluorescence
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