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. Author manuscript; available in PMC: 2022 Dec 1.
Published in final edited form as: J Neurosci Methods. 2021 Sep 17;364:109358. doi: 10.1016/j.jneumeth.2021.109358

Figure 3. Enzymatic dissection of radial nerve cords (RNCs).

Figure 3.

Strips of body wall containing the RNCs were treated with collagenase for A) 8-, B) 18-, C) 24-, D) 42- and E) 48-hours. A) After digestion for 8 hours in collagenase, the RNC (delineated by stippled lines) is still surrounded by connective tissue and other body wall components. B) The RNC is visible after an 18-hour digestion in collagenase but still shows significant debris associated with it. C) RNCs with peripheral nerves (arrowhead) after 24-hr digestion in collagenase show very little signs of surrounding debris. RNCs have a tendency to curl-up when free of surrounding tissue. D) RNC after a 42-hour or E) 48-hour digestion shows that the expected structure and morphology remains intact. Peripheral nerves are observed (arrowhead). (F-I) RNC cross-section immunostaining using anti-collagen immunohistochemistry. F) Immunohistochemical studies on tissue sections of the body wall and RNC show the collagen (arrows) that separates the RNC from the surrounding body wall for in vivo cross sections. The thin band of connective tissue that separates the ectoneural (Ec) component from the hyponeural (Hy) is also observed (arrowhead). Following collagenase dissection and culture in vitro for G) 3-, H) 7- and I) 11-days little collagen immunoreactivity is associated with the RNC explants. Scale bar = 10 μm in A-E and 20 μm in F-I.