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(+)JQ1 inhibits microglial synaptic elimination in both transgenic zebrafish and rat primary culture models expressing human 4R/0N-Tau. a Whole mount immunofluorescence for PSD95-immunoreactive post-synaptic puncta in the telencephalon (above) and optic tectum (below; approximate regions of images shown in low-magnification panel to right). Labeled brains were imaged by confocal microscopy. Maximum intensity projections are shown. PSD95-immunoreactive post-synaptic puncta were quantified in tissue volumes from (b) optic tectum and (c) telencephalon using unbiased automated image analysis algorithms. Data points show individual zebrafish (n = 17–43 zebrafish/group), bars show mean ± SE. ****p < 0.0001, ***p < 0.001, *p < 0.05, 1-way ANOVA with Šidák multiple comparisons test. d Fixed whole mount brains from [Tau or Ctrl] x Tg(mpeg1:egfp) zebrafish immunolabeled for GFP and PSD95 at 4dpf and imaged by confocal microscopy. Alpha shaded 3D projections of microglia are shown. The inset shows a single confocal plane from a Tau zebrafish, illustrating PSD95-immunoreactive synaptic material inside the cytoplasm of the microglial cell (arrow). e PSD95+ puncta quantified inside microglia in Ctrl, Tau, and [Tau + (+)JQ1] zebrafish as shown in panel d. Data points show individual microglia (n = 52–83 microglia from 6 zebrafish/group), bars show group mean ± SE. ****p < 0.0001, 1-way ANOVA with Tukey multiple comparisons test. f Primary embryonic rat cortex cultures were established under conditions promoting microglial survival and differentiation. Neurons were transfected to express human 0N/4R-Tau and membrane-anchored GFP (mGFP), or mGFP alone. Cultures were exposed to 1 μM (+)JQ1 (or vehicle only) and labeled for Iba1 (microglia; red), mGFP (transfected neurons; green), PSD95 (synaptic puncta; white), and DAPI (nuclei; blue). g PSD95+ puncta were quantified inside microglia (Supplementary Fig. S32) from each of the primary rat cortex cultures. Data points show individual microglia (n = 65–87 microglia from 3 biological replicates per group), bars show group mean ± SE. ****p < 0.0001, 1-way ANOVA with Šidák multiple comparisons test. Source data are provided as a Source Data file. Exact p-values shown in Supplementary Table 21.
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