Rapid Redistribution of Synaptic PSD-95 in the Neocortex In Vivo (original) (raw)
Figure 2
Rapid Turnover of PSD-95 in Individual PSDs
(A) Schematic of the experimental setup. One laser (λ ~ 1,030 nm, magenta) was used to excite mCherry and photoactivated paGFP (paGFP*). A second laser (λ ~ 810 nm, blue) was used for photoactivation within a region of interest (blue box) within the field of view (magenta box). BS, beam-splitting cube; DM, dichroic mirror; OB, objective; R, G, photomultiplier tubes; SM, scan mirror.
(B) Schematic of the experiment. Spines were selected for photoactivation (pre-pa, blue box), followed by time-lapse imaging.
(C) Images collected before, and after (0 min, 90 min, and 24 h) photoactivation of two spines on different dendrites (age, P17).
(D) Time course of PSD-95-paGFP* fluorescence (same as in [C]). The initial portion of the decay (inset) was used to extract the retention time (_τ_r) of PSD-95-paGFP*.
(E) Retention times. Circles represent single spines and the horizontal bars indicate the medians (_τ_median). (P10–P12: _τ_median = 30 min, range 4–77 min, n = 6 animals, 74 spines; P13–P15: _τ_median = 34 min, range 5–108 min, n = 6 animals, 108 spines; P16–P18: _τ_median = 37 min, range 9–194 min, n = 8 animals, 95 spines; P19–P30: _τ_median = 43 min, range 11–291 min, n = 8 animals, 59 spines; >P60: _τ_median = 106 min, range 20–289 min, n = 4 animals, 25 spines).