Extended Data Fig. 10: The SNS promotes atherosclerosis during ageing.
From: Neuroimmune cardiovascular interfaces control atherosclerosis
a, Spearman correlation coefficients of SSNA vs plaque volume in aortic arch and abdominal aorta in young, adult and aged Apoe−/− mice (n = 26 mice). b, Representative raw signals of SSNA in a time window of 10 min and relative quantification of SSNA spikes in adult and aged Apoe−/− mice before and after Coeliac vagotomy (CVNX) (n = 6 adult and 7 aged). c, Levels of neuron activation-related genes in LCM-derived sympathetic PvaGs in aged WT versus Apoe−/− mice (n = 5 WT PvaGs, 6 Apoe−/− PVaGs). Signal intensities and statistics are reported in supplementary Table 6. d, Approach to 4 weeks 6-OHDA-induced chemical sympathetic denervation in aged Apoe−/− mice. e, TH+ neurons in the locus coeruleus (n = 5 control and 5 OHDA). f, Analysis of spleen noradrenaline (n = 4 control and 7 OHDA), splenic TH+ area (n = 4 control and 4 OHDA), and aortic root TH+ area (n = 3 control and 3 OHDA). g,h, Effect of 6-OHDA denervation on plaque and ATLO. g, OR/H stained abdominal aorta showing ATLO cellularity. (h, Quantification of abdominal aorta media area; macrophage area (CD68+), necrotic core area, SMC area (SMA+), collagen area (Sirius red+), fibrous cap thickness in plaque; and CD3e+Foxp3+ T regulatory cells in ATLO ; n = 4 control and 4 OHDA. i, Measurement of serum cholesterol (n = 6 control and 8 OHDA), relative organ weight (n = 5 per group). j, Flow cytometry gating strategies and enumeration of Lin-Sca1+Kit+CD150+CD48- haematopoietic stem cells (HSC) gated from LSK (Lin-Sca1+Kit+) or Lin-Sca1-Kit+CD34+CD16/32+ granulocyte-macrophage progenitors (GMP) gated from myeloid progenitor cells (MPC) (Lin-Sca1+Kit-) in the bone marrow from total live cells (n = 5 control and 5 OHDA). (k, Gating strategies and quantification of CD11c+CD11b+ myeloid cells and CD4+Foxp3+ T regulatory cells in spleen and RLN from total live cells (n = 5 control and 5 OHDA). l, Approach to 8 months CGX selective surgical denervation in adult Apoe−/− mice. m, Analysis of spleen noradrenaline (n = 4 sham and 5 CGX), aortic root TH+ area (n = 4 sham and 5 CGX), and root plaque size (n = 4 sham and 7 CGX). n, Changes in serum cholesterol (n = 3 sham and 9 CGX); relative organ weights after 8 months of surgery (n = 4 sham and 9 CGX). o, Flow cytometry gating strategy and quantification of number of CD11b+ myeloid cells and CD4+Foxp3+ T regulatory cells in spleen and RLN from total live cells (n = 4 sham; 4 CGX). p,q, Effect of surgical denervation on ATLO and plaque cellularity. p, Histological staining shows ATLO cellularity., q, Quantification of abdominal aorta media area; macrophage area (CD68+), necrotic core area, SMC area (SMA+), collagen area (Sirius red+), fibrous cap thickness in plaque; and CD3e+Foxp3+ T regulatory cells in ATLO ; n = 4 sham, 5 CGX. r, Analyses of internal diameter and ß-stiffness in ascending aorta and abdominal aorta of sham and CGX- mice before surgery (Basal) and 8 months after surgery. s, Systolic and diastolic blood pressure measurement (Basal) and at every 2 months up to 8 months after surgery n = r,s: 7 sham, 10 CGX before surgery, and 5 sham, 9 CGX at 8 months post-CGX.For e, scale bars, 20 µm; for g,p, scale bars 50 µm. Data are means ± s.e.m. n represents biologically independent animals. Two-way ANOVA with Bonferroni post hoc test (b,j,k,o); two-sided unpaired Student´s t-test (e,f,h,i,m,n,q); multiple unpaired t-test corrected for multiple comparisons (Bonferroni) (c,i,n); mixed-model ANOVA with Bonferroni post hoc test (r,s). t, Comparison of short-term (4 weeks) pharmacological deletion of the SNS and long-term (8 months) outcome of CGX on plaque and ATLO
