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Supplemental Data for Ali et al., 2021

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posted on 23.07.2021, 17:29 by Maria F. Ali, Andrew J. Latimer, Yinxue Wang, Leah Hogenmiller, Laura Fontenas, Adam J. Isabella, Cecilia Moens, Cecilia B. Moens, Guoqiang YuGuoqiang Yu, Sarah Kucenas
Supplemental figures and movies for Ali et al., 2021.

Figure S1. Met inhibition alters the distance that dorsal OPCs migrate. (A-F) Quantifications taken from time-lapse movies of DMSO (n = 7) and MK2461-treated (n = 7) larvae in (Figure 1C). Statistical test: Student’s t-test.

Figure S2. met mutants exhibit a 16 bp insertion and reduced dorsal OLCs. (A) Bright-field images of met+/+, met+/uva38, and metuva38/uva38 siblings at 3 dpf reveal no developmental delay in metuva38/uva38 larvae. (B) RT-PCR of mRNA transcripts extracted from 48 hpf met+/+, met+/uva38, and metuva38/uva38 embryos demonstrating 16 bp insertion in met+/uva38 and metuva38/uva38 RNA transcripts. (C) Images of 72 hpf olig2:egfp met+/+ and metuva38/uva38 larvae spinal cords. Yellow open arrowheads denote dorsal OLCs. Dashed yellow line denotes ventral edge of the spinal cord. (D) Quantifications taken from spinal cord images of met+/+ (n = 3), met+/uva38 (n = 6), met+/fh534 (n= 7), and metuva38/fh534 (n = 6) larvae. Mean with SEM. Statistical test: 1-way ANOVA with Tukey’s Multiple Comparison Test. Scale bars, 0.5 mm (A), 20 μm (C).

Figure S3. met mutants do not exhibit defects in OPC distance traveled or velocity. (A-G) Quantifications taken from 18 hour time-lapse movies of 55 hpf olig2:egfp met+/+ (n = 8), met+/uva38 (n = 8), and metuva38/uva38 (n = 6) larvae in (4C). Statistical test: 1-way ANOVA with Tukey’s Multiple Comparison Test.

Figure S4. hgfa mutants exhibit reduced OPC numbers. (A) Lateral (upper) and optical section (lower) views of Sox10 antibody labeled 72 hpf wildtype, hgfa+/+, and hgfafh528/fh528 larvae. Horizontal yellow line denotes location of orthogonal view. Yellow dashed line denotes ventral edge of the spinal cord. Yellow open arrowheads denote sox10+ OLCs. Dashed yellow circle denotes boundary of spinal cord. (B-E) Quantifications taken from images of 72 hpf Sox10 antibody labeled hgfa+/+ (n = 14), hgfa+/fh528 (n = 11), and hgfafh528/fh528 (n = 14) larval spinal cords. Mean with SEM. Statistical test: 1-way ANOVA with Tukey’s Multiple Comparison Test. Scale bar, 10 μm.

Figure S5. met mutants exhibit wildtype OPC specification. (A) Transverse sections of 48 hpf olig2:egfp;sox10:mrfp; met+/+ and metuva38/uva38 embryos. Open yellow arrowheads denote sox10+/olig2+ OPCs. Dashed yellow circle denotes boundary of spinal cord. (B-E) Quantifications of olig2+/sox10+ OPCs from serial sections of 48 hpf olig2:egfp;sox10:mrfp met+/+ (n = 6) and met+/uva38 (n = 4), and metuva38/uva38 (n = 4) embryos. Mean with SEM. Statistical test: 1-way ANOVA with Tukey’s Multiple Comparison Test. Scale bar, 10 μm.

Movie 1. New cell tracking software labels migratory olig2+ cells in olig2:egfp larvae from 55 to 76 hpf. Images were taken every 10 minutes and the movie runs at 10 frames per second (fps).

Movie 2. DMSO-treated olig2:egfp larvae exhibit wildtype OPC migration from 55 hpf to 76 hpf. Images were taken every 10 minutes and the movie runs at 10 fps.

Movie 3. MK2461-treated olig2:egfp larvae exhibit reduced dorsal OPC migration from 55 to 76 hpf. Images were taken every 10 minutes and the movie runs at 10 fps.

Movie 4. olig2:egfp;met+/+ larvae exhibit wildtype OPC migration from 55 to 76 hpf. Images were taken every 10 minutes and the movies runs at 10 fps.

Movie 5. olig2:egfp;metuva38/uva38 larvae exhibit reduced dorsal OPC migration from 55 to 76 hpf. Images were taken every 10 minutes and the movie runs at 10 fps.


History

Article title

Met is required for oligodendrocyte progenitor cell migration in Danio rerio