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. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: J Neurosci. 2012 Jan 25;32(4):1496–1506. doi: 10.1523/JNEUROSCI.4176-11.2012

Figure 2. Motor neuron subtype specific, protein synthesis independent response to ephrin-A5.

Figure 2

(A) RALDH2 is expressed by ES-derived LMC motor neurons ((Foxp1+; Hb9+) – Arrow) and not by other motor neuron subtypes ((Foxp1−; Hb9+) – Arrow head). (Scale bar: 10 μm) (B) Immunostaining on mouse embryo section (E12.5) for RALDH2 (Red) shows that the enzyme specifically labels LMC axons projecting toward the forelimb but is not present in other motor branches. (SC : Spinal Cord - DRG: Dorsal Root Ganglia – LMC : Lateral Motor Column - Limb Br. : Limb axonal branch (LMC axons) – Axial Br.: Axial axonal branch. scale bar : 100 μm. (C) RALDH2 is observed in a subset of axons growing from ES-derived brachio-thoracic motor neurons plated on laminin substrate. (D) ephrin-A5 at 50 ng/ml induces preferentially the collapse of non-LMC motor axons (HB9-GFP+ / RALDH2− axons) compare to LMC axons (HB9-GFP+ / RALDH2+ axons) (n=3) while 300 ng/ml leads to similar collapse of both populations. (Scale bar: 15 μm). (E) Dose-response of brachio-thoracic ES-MNs to ephrin-A5 in absence or presence of cycloheximide. All significantly active concentrations of ephrin-A5 (starting at 30 ng/ml ; n=3) induce collapse in a protein synthesis independent manner. (Two-way ANOVA, n.s. = no significance, error bar: STD).