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. Author manuscript; available in PMC: 2017 Feb 16.
Published in final edited form as: Nat Methods. 2016 Jun 6;13(8):673–678. doi: 10.1038/nmeth.3894

Figure 4. GFE3 expression in zebrafish spinal cord results in Gephyrin ablation and behavioral deficits.

Figure 4

(a) A confocal image of a live zebrafish age 2 days post fertilization (dpf) shows mosaic expression of GFE3-GFP driven by an enhancer element of the mnx1 gene that restricts expression to spinal motoneurons. (b,c) Immunostaining of Gephyrin (magenta) and GFP (green) in motoneurons expressing RandE3-GFP (b) or GFE3 (c). Scale bar represents 5 µm. (d) Gephyrin levels in cells expressing RandE3-GFP (64 ± 5 a.u.; n = 5 fish, 6 cells per fish, 3 independent experiments) and in cells expressing GFE3-GFP (23 ± 2 a.u.; P < 0.008, Mann-Whitney; n = 5 fish, 6 cells per fish, 3 independent experiments). (e) Zebrafish expressing RandE3-GFP, age 24 hours post fertilization, exhibit normal spontaneous tail flicking, including touching tip of the tail to the base. (f) Zebrafish expressing GFE3-GFP in motoneurons are unable to curl their tails sufficiently to touch the tip to the base. (g) Quantitation of the minimum distance between the base of the tail and the tip, measured during 5 spontaneous flicks, reveals a > 100% increase in zebrafish that express GFE3-GFP vs. those expressing RandE3-GFP (P < 0.003, Mann-Whitney; n = 5 fish, 5 flicks per fish). Bars indicate means ± SEM.