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. 2022 Jan 28;15:751850. doi: 10.3389/fncir.2021.751850

Figure 6.

Figure 6

Retrograde labeling of central EVN neurons. Fluoro Gold fluorescent retrograde tracer was injected unilaterally in the inner ear through the posterior semicircular canal and the efferent neurons were traced back retrogradely in the EVN on both sides. (A) Dissected bony inner ear following Fast Green dye injection through the posterior semicircular canal, arrow—injection site, PC, posterior semicircular canal, CC, common crus; AC, anterior crista; HC, horizontal crista. (B,C) Maximum projection image from z-stacks of the triad and posterior crista following dissections of a Fluoro Gold injected inner ear, dashed line outlines the vestibular cristae. curved dashed line—utricular striola, AC, anterior crista; HC, horizontal crista; U, utricle; arrow—eminentia cruciata; arrowheads—fluorescent neuroepithelium. Scale bar (B): 500 μm, Scale bar (C): 100 μm. (D) Photo-merged image of transgenic Chat-gCaMP6f mouse used for retrograde tracing and EVN cell counting. The ChAT dependent GCaMP6fastexpression was visualized by GFP immunolabeling in green. 4V—4th ventricle, MVN—medial vestibular nucleus, EVN—Efferent Vestibular Nucleus, abd—abducens nucleus, g7n—genu of the facial nerve, 7n—facial nucleus. Scale bar: 1 mm. (E,F) Left and Right side EVN nuclei under higher magnification. abd—abducens nucleus, 7n—genu of the facial nerve. Scale bar: 50 μm. (G) Fluoro Gold (FG) labeling of the EVN. 4V—4th ventricle, EVN—Efferent Vestibular Nucleus, arrows—position of the EVN, abd—abducens nucleus, 7n—genu of the facial nerve. Scale bar: 500 μm. (H,I) FG labeling in the left side EVN and the composite image of GFP and FG labeling. Arrow points to GFP but not FG-positive EVN neuron. Scale bar: 50 μm. (J,K) FG labeling in the right side EVN and the composite image of GFP and FG labeling. Arrow points to GFP-positive, FG-negative EVN neuron. Scale bar: 50 μm.