Fig. 1: Mitochondria display highly compartmentalized, layer-specific, morphology in dendrites of CA1 PNs in vivo. | Nature Communications

Fig. 1: Mitochondria display highly compartmentalized, layer-specific, morphology in dendrites of CA1 PNs in vivo.

From: Activity-dependent compartmentalization of dendritic mitochondria morphology through local regulation of fusion-fission balance in neurons in vivo

Fig. 1

a Schematic of in utero electroporation (IUE) procedure used to sparsely express fluorescent reporters, cDNAs or shRNAs in CA1 PNs of the hippocampus. b Schematic of single cell electroporation (SCE), used to express fluorescent reporters into single adult CA1 pyramidal neurons in the hippocampus. This method allows for rapid expression of plasmid DNA and in vivo two-photon visualization of individual neurons in an adult anesthetized mouse in vivo. Portions of (a) and (b) were created with BioRender.com. c Representative image of a single CA1 PN expressing a mitochondrial matrix reporter (mt-YFP) via SCE following fixation and re-imaged using confocal microscopy on a single vibratome section. Scale bar, 50 μm. d High-magnification representative images of dendrites from the three hippocampal compartments—basal (SO), apical oblique (SR), and apical tuft (SLM)—expressing a mitochondrial matrix marker (mt-YFP or mt-DsRed) imaged either in vivo using two-photon microscopy (left) or post-fixation with confocal microscopy (right). Scale bar, 5 μm. e–g Quantification of individual mitochondrial length (e), mitochondrial segment occupancy (f), and intra-segment mitochondrial length variability (g) from basal, apical oblique, and apical tuft dendrites captured in vivo. (Basal: n = 422 mitochondria; n = 37 dendritic segments; mean length = 3.621 μm ± 0.124 (SEM); mean occupancy = 56.11% ± 2.49% (SEM); mean intra-segment variability = 2.420 μm ± 0.198 (SEM); Apical Oblique: n = 263 mitochondria; n = 35 dendritic segments; mean length = 4.719 μm ± 0.249 (SEM); mean occupancy = 68.82% ± 2.20% (SEM); mean intra-segment variability = 3.185 μm ± 0.370 (SEM); Apical Tuft: n = 90 mitochondria; n = 23 dendritic segments; mean length = 13.02 μm ± 1.041 (SEM); mean occupancy = 89.26% ± 1.24% (SEM); mean intra-segment variability = 8.018 μm ± 6.359 (SEM)). h–j Quantification of individual mitochondrial length (h), mitochondrial segment occupancy (i), and intra-segment mitochondrial length variability (j) from basal, apical oblique, and apical tuft dendrites following fixation. (Basal: n = 313 mitochondria; n = 32 dendritic segments; mean length = 1.065 μm ± 0.029 (SEM); mean occupancy = 33.01% ± 1.84% (SEM); mean intra-segment variability = 0.4736 μm ± 0.030 (SEM); Apical Oblique: n = 184 mitochondria; n = 28 dendritic segments; mean length = 1.569 μm ± 0.104 (SEM); mean occupancy = 42.38% ± 2.98% (SEM); mean intra-segment variability = 0.9169 μm ± 0.149 (SEM); Apical Tuft: n = 226 mitochondria; n = 33 dendritic segments; mean length = 6.562 μm ± 0.437 (SEM); mean occupancy = 84.19% ± 1.44% (SEM); mean intra-segment variability = 5.695 μm ± 0.552 (SEM)). p values are indicated in the figure following a one way ANOVA with Sidak’s multiple comparisons test. Data are shown as individual points on box plots with 25th, 50th and 75th percentiles indicated with whiskers indicating min and max values.

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