Fig. 4: Kdm4a and Trpm7 significantly regulate the size of mossy fiber boutons.
From: Kdm4a is an activity downregulated barrier to generate engrams for memory separation
a Schematic of sparse labeling of the mouse DG GCs using shKdm4a or shCtrl lentivirus. Scale bar, 5 μm. b Dendritic spine density are not significantly different between shKdm4a (n = 3, dendrites = 34) and shCtrl (n = 3, dendrites = 26) mice. Two-tailed unpaired t-test, t58 = 1.142, p = 0.2583. The density of mushroom-like dendritic spines are similar between shKdm4a (n = 3, dendrites = 10) and shCtrl (n = 3, dendrites = 10). Two-tailed unpaired t-test, t18 = 0.01855, p = 0.9854. The proportion of mushroom-like dendritic spines are not significantly different between shKdm4a (n = 3, dendrites = 10) and shCtrl (n = 3, dendrites = 10). Two-tailed unpaired t-test, t18 = 1.109, p = 0.2818. Data are presented as mean ± s.e.m. c The width of dendritic spines are similar between shKdm4a (n = 3, spines = 265) and shCtrl (n = 3, spines = 265). Two-tailed unpaired t-test, t528 = 0.9885, p = 0.3233. Right: Frequency distribution plot of dendritic spine width. Data are presented as mean ± s.e.m. d Schematic illustration of subregions of mouse dentate gyrus and CA3. Granule cell layer: gcl, molecular layer: ml stratum pyramidalis: sp stratum lucidum: sl stratum radiatum: sr. e Immunofluorescence staining of EYFP+ neuron axonal projections in the stratum lucidum of CA3. Scale bar, 2 μm. f The size of DG MFb was significantly increased in the shKdm4a mice (n = 3, boutons = 117) compared with shCtrl control mice (n = 3, boutons = 100). Two-tailed unpaired t-test, t214 = 5.333, **** p < 0.0001. (Right) Frequency distribution histogram of DG mossy fiber bouton size. Data are presented as mean ± s.e.m. g Illustration of CRISPR-a based transcription activation system for overexpression of Trpm7. n = 3, One-way ANOVA followed by Bonferroni test, F3, 8 = 53.28, p < 0.0001. ** p = 0.003, **** p < 0.0001. Data are presented as mean ± s.e.m. h Immunofluorescence staining of EGFP+ neuron axonal projections in the stratum lucidum of CA3. Scale bar, 5 μm. i, The area of DG MFb in the sgTrpm7 mice (n = 3, boutons = 96) was significantly larger than that in the sgCtrl mice (n = 3, boutons = 96). Two-tailed unpaired t-test, t190 = 4.280, **** p < 0.0001. Right: Frequency distribution histogram of mossy fiber bouton size. Data are presented as mean ± s.e.m.
