Fig. 5
Mechanical loading regulates sympathetic activity through hypothalamus AgRP neurons. a Diagram of the AAV9-hSyn-GFP injection sites in the ARC area of the WT mice. b Representative images of GFP+ neurons in the PVN of the hypothalamus AAV9-hSyn-GFP injection in ARC. c Diagram of the CTB injection site in the PVN area of the WT mice. d Representative images of GTB+ neurons in the ARC of the hypothalamus after CTB injection in PVN. Representative images of immunofluorescence staining e and quantitative analysis of the pCREB (red) and CTB (green) f in the ARC of the hypothalamus of WT mice underwent three consecutive days of axial compression loading of tibiae or control sham load after CTB injection in the PVN for 5 days. Scale bar, 50 μm. AgRP-Ires-Cre mice injected with pAAV-hSyn-DIO-hM4d(Gi)-mCherry in one side of the ARC (right) and control AAV in the other side (left). Mice injected saline or CNO (0.3 mg/kg of body weight, i.p.) before loading. Representative images of immunofluorescence staining g and quantitative analysis h of the pCREB (green) and mCherry (red) in the ARC of hypothalamus of AgRP-Ires-Cre mice underwent three consecutive days of axial compression loading of tibiae or control sham load. Scale bar, 50 μm. Representative images of immunofluorescence staining i and quantitative analysis of the TH (green) j in the PVN of hypothalamus of AgRP-Ires-Cre mice that underwent three consecutive days of axial compression loading of tibiae. Scale bar, 50 μm. n ≥ 5 per group. *P < 0.05, **P < 0.01, and N.S. indicates not significant. Statistical significance was determined by Student’s t-test
