Figure 5

<p>(A) Changes in the density of intramuscular nerves in muscle tissue of mice 3 and 28 days after injection of either PBS, <i>B. asper</i> venom, Mtx or BaP1. Axons in nerves were visualized by immunostaining with a mouse anti-human neurofilament protein, as described in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019834#s2" target="_blank">Methods</a>. The number of intramuscular nerves per mm² of tissue area showing at least one immunostained axon was quantified. Results are presented as mean±SD (n = 5). A significant drop (* p < 0.05) in nerves per area was observed at 3 days in samples injected with either venom, Mtx or BaP1, as compared to samples injected with PBS, whereas no differences in the number of nerves per area were detected between treatments at 28 days. (B) Axonal density in intramuscular nerves 28 days after injection of the various agents. The number of axons within each nerve was determined and expressed in terms of axons per nerve area. Results are presented as mean±SD (n = 11). * p < 0.05 when compared with axonal density in control muscles injected with PBS. **p < 0.05 when compared with axonal density in muscles injected with Mtx. (C to F) Light micrograph sections of mouse muscle tissue collected 28 days after injection of (C) PBS, (D) <i>B. asper</i> venom, (E) Mtx, and (F) BaP1. Sections were immunostained for neurofilament protein to detect axons in nerves (arrows). Notice the evident drop in the number of axons in samples from tissue injected with venom or BaP1. Bar represents 50 µm.