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. 2018 Mar 8;27(10):1754–1762. doi: 10.1093/hmg/ddy083

Figure 5.

Figure 5.

GR expression results in reduction of motor neuron length with increased cell death. (A) Example images of GFP-positive motor neurons in 48 hpf zebrafish larvae expressing GR DPR, and a non-transgenic larva as a control. Axons of motor neurons in GR100-expressing larvae appear shorter and less branched. (B) Measurement of the length of motor neurons in the spinal cord showed a significant decrease in motor neuron length when GR DPR is expressed (N = 2, measurement performed from seven embryos per condition, five ventral roots per embryo; two-tailed P < 0.0001 calculated by unpaired t-test). (C). Measurement of the length of motor neurons in the spinal cord before expression of the GR DPR at 48 hpf and at 80 hpf after 24 h of GR100 expression showed a significant decrease in motor neuron length only when GR DPR is expressed (N = 2, measurement performed from seven embryos per condition, five ventral roots per embryo; before heat shock group: two-tailed P = 0.7693 calculated by unpaired t-test; after heat shock group: two-tailed P < 0.0001 calculated by unpaired t-test). (D) Examples of GFP-positive motor neurons in 48 hpf and 80 hpf zebrafish larvae (prior to and after expressing GR DPR), with non-transgenic larvae served as a control. Axons of motor neurons abnormal after GR100 expression. (E) Examples of acridine orange–stained zebrafish larval spinal cords expressing GR DPR in neurons with larvae not expressing the DPR was used as a control. HuC expression was used to set limits while imaging, boundary of the cord is marked with black dotted lines. GR100 DPR expressing larvae show visibly more acridine orange positive cells (black dots, indicated by arrowheads), many of which located in the ventral region of the spinal cord. (F) Quantification of the number of acridine orange positive cells/larva/field in the spinal cord of larvae expressing GR DPR showed significantly greater number of AO positive cells in the spinal cord in comparison to larvae not expressing GR DPR (n = 12 larvae per genotype; two-tailed P = 0.0068 calculated by unpaired t-test).