Fig 1: Eph receptor inhibition does not alter auditory vs. vestibular targeting.(A) RDA tracing combined with neurofilament immunofluorescence was used to determine whether auditory axons were correctly targeted. An example of EphB1-Fc treated embryo with normal targeting is shown. (B) Example of EphA3-Fc treated embryo with normal auditory targeting. (C) Similarly, treatment with EphA4-Fc did not alter central targeting of auditory VIIIth nerve axons. (D) Targeting of vestibular axons was determined in embryos that showed GFP transfection in vestibular, but not auditory, ganglion cells. Axons were followed through brainstem sections. Transfection in this example was with EphA4; all GFP labeled axons were found in vestibular projection regions. (E) Transfection with EphB2 plasmid did not alter central vestibular VIIIth nerve projections. (F) Misexpression of kiEphA4 did not alter central vestibular projections. (G) Similarly, kiEphB2-transfected embryos showed no mistargeting of vestibular axons into auditory projection regions. Scale bar, 100 µm in A-C and 300 µm in D-G.
Fig 2: Eph receptor inhibition causes reduction in hindbrain compartment.(A-D) Neurofilament immunofluorescence (left) of hindbrain sections at level of VIIIth nerve entry with auditory (black) and vestibular (gray) components schematized (right). Red dashed line indicates entire dorsoventral extent of the VIIIth nerve projections in hindbrain. Volumes found in IgG-Fc (A) EphB1-Fc (B), and EphA3-Fc (C) did not significantly differ. (D) Vestibular hindbrain volume is reduced following EphA4-Fc treatment. (E,F) Graphs of mean auditory (E) and vestibular (F) volumes for all treatment groups. Numbers of samples are indicated in each bar; asterisk indicates p <0.05.
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