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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Nat Neurosci. 2014 Nov 2;17(12):1830–1839. doi: 10.1038/nn.3854

Figure 5. Quantitative intra-axonal organelle profile of ARCAGRP→PVH and ARCPOMC→PVH boutons.

Figure 5

a, Number of SVs plotted against LVs for each bouton. b,c, Number of SVs and LVs for each bouton with respect to presence of a postsynaptic contact site. (b) Small vesicles. AGRP non-synaptic: 38.2 ± 9.4, n = 27, AGRP synaptic: 365 ± 52, n = 24, U-test, U = 17.5, P < 0.001; POMC non-synaptic: 43.1 ± 11.3, n = 11, POMC synaptic: 650 ± 77.3, n = 40, U-test, U = 0.0, P < 0.001. n = 1 mouse each. (c) Large vesicles. AGRP non-synaptic: 62.2 ± 7.2, n = 27, AGRP synaptic: 86.1 ± 12.9, n = 24, U-test, U = 261.5, P = 0.24; POMC non-synaptic: 61.6 ± 17.4, n = 11; POMC synaptic: 30.5 ± 5.0, n = 40, U-test, U = 168.5, P = 0.24. n = 1 mouse each. d–f Number of mitochondria in each bouton with respect to cell type (d) (AGRP: 0.83 ± 0.1; POMC: 1.23 ± 0.14, n = 51 boutons each, n = 1 mouse each; U-test, U = 1007.5, P = 0.03), SV (e), and LV (f) counts. Red, AGRP. Blue, POMC. Black, regression line. g,h, Distribution (g) and mean number (h) of mitochondria in each bouton with respect to the presence of a postsynaptic partner (non-synaptic: 0.77 ± 0.12 mitochondria, n = 43 boutons; synaptic: 1.3 ± 0.15 mitochondria, n = 47 boutons; pooled from one Agrp-Cre and one Pomc-Cre mouse, U-test, U = 689.5, P = 0.005). i, Example reconstructions of axonal segments with multiple GESEM-labeled boutons. a1 and a2: AGRP axons with multiple boutons, one makes a synaptic contact and the others are non-synaptic and contain mostly LVs. p1: A POMC axon making two synaptic contacts onto two dendritic segments. p2: A POMC axon with two boutons filled with LVs and no apparent SVs or postsynaptic contacts. Scale, 1 μm. j, SV and LV numbers in individual boutons, where boutons that are on the same axonal segment are connected with a line. Green highlight: axons reconstructed in (i). k, Vesicle numbers in adjacent boutons can be dramatically different for SVs (left) but LV numbers are usually similar (right). l, From ARCAGRP→PVH axons, SV and LV numbers in adjacent boutons with respect to the presence of a postsynaptic element (small vesicles, non-synaptic: 48.8 ± 14.8, synaptic: 479.6 ± 112, paired t-test, t(7) = −3.77, P = 0.007; large vesicles, non-synaptic: 69 ± 11.9, synaptic: 115.9 ± 32, paired t-test, t(7) = −1.82, P = 0.11; n = 8 AGRP axonal segments from one mouse). Values are mean ± s.e.m. n.s. P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001.