Endothelin-1 regulates cardiac sympathetic innervation in the rodent heart by controlling nerve growth factor expression
J. Clin. Invest. Masaki Ieda, et al. 113:876 doi:10.1172/JCI19480 [
Go to this article.]
Figure 5Edn1–/– embryos display a loss of SG neurons due to excess apoptosis. (A)
Edn1+/+ and
Edn1–/– whole-mount embryos at E12.5 were immunostained with anti-TH antibody. SCG, superior cervical ganglion; fl, forelimb. Similar results were obtained from four separate experiments. (B–E) TH immunostaining, cresyl violet staining (CV), Ki-67 immunostaining, and TUNEL staining of
Edn1+/+ and
Edn1–/– SG at E12.5, E15.5, and E18.5 at the same level of section. Note that at E18.5,
Edn1–/– SG were considerably smaller than
Edn1+/+ SG and increased apoptosis was detected. (F–H) Time course of the number of neurons, Ki-67
+ cells per 100 neurons, and TUNEL
+ cells per 1,000 neurons in SG was shown (
n = 5). (I) Time course of NGF expression in
Edn1+/+ and
Edn1–/– hearts was determined by quantitative RT-PCR (
n = 3). (J and K) Immunostaining for TH in the heart of
Edn1+/+ and
Edn1–/– embryos at E15.5 and E18.5. TH-immunopositive nerve fibers were slightly detected from E15.5. LA, left atrium; LV, left ventricle. The immunopositive nerve areas for TH were determined using NIH Image. (
n = 4.) *
P < 0.001; **
P < 0.01;
#P < 0.05. NS, not significant vs. relative control. Scale bar: 500 μm (A), 100 μm (B and J), 10 μm (C and D), 50 μm (E). Black bars,
Edn1+/+; white bars,
Edn1–/–.
