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. 2022 Aug 23;11:e74295. doi: 10.7554/eLife.74295

Figure 2. Neuronal defects in norepinephrine (NE) biosynthesis, metabolism, and survival in satellite glia-depleted mice.

(A) Tyrosine hydroxylase (TH) expression is downregulated in BLBP:iDTA sympathetic neurons. Insets also show atrophied neuronal cell bodies in mutant ganglia compared to controls. Scale bar: 100 μm for upper panels and 30 μm for insets. (B) Transcripts for Th and DBH, key enzymes in norepinephrine biosynthesis, are decreased in BLBP:iDTA SCG relative to control ganglia. Data are presented as means ± SEM from superior cervical ganglia (SCG) collected from n = 3–4 animals per genotype. ***p<0.001, ****p<0.0001, t-test with Bonferroni–Dunn’s correction. (C) Histogram shows a greater distribution of smaller soma sizes in mutant neurons compared to controls. Results are means ± SEM from n = 6–8 animals per genotype, **p<0.01, two-way ANOVA with Bonferroni’s correction. (D) Reduced soma sizes, represented as median values for soma areas (μm2), of sympathetic neurons from BLBP:iDTA mice compared to controls at 5 dpi. Values are means ± SEM from n = 8 control and 6 mutant animals, *p<0.05, t-test. (E) Immunostaining shows reduced p-S6 levels in BLBP:iDTA ganglia. Scale bar:100 μm. (F, G) Cell counts in Nissl-stained SCG tissue sections show reduced sympathetic neuron numbers in satellite glia-depleted mice 14 dpi. Results are means ± SEM from n = 4 control and 5 mutant animals. **p<0.01, t-test.

Figure 2—source data 1. Raw data for neuronal morphology and survival.

Figure 2.

Figure 2—figure supplement 1. Neuronal morphology and signaling in BLBP:iDTA mice.

Figure 2—figure supplement 1.

(A) Immunostaining shows reduced phospho-4E-BP1 expression in BLBP:iDTA sympathetic neurons compared to controls at 5 dpi. Scale bar: 100 μm. (B) Sympathetic neuron numbers in satellite glia-depleted mice are similar to control values at 5 dpi. Results are mean ± SEM from n = 5 control and 4 mutant animals, n.s., not significant, t-test. (C) Sympathetic axon innervation of the heart is unaffected in adult BLBP:iDTA mice based on iDISCO-based tissue clearing and wholemount tyrosine hydroxylase (TH) immunostaining. Scale bar: 100 µm. (D, E) Quantification of axon length and branches in the heart in BLBP:iDTA and control mice. Data are mean ± SEM from n = 4 control and 5 mutant animals, n.s., not significant, t-test.
Figure 2—figure supplement 1—source data 1. Raw data for neuronal morphology and survival.