ZFIN Figure: Herbert et al., 2017, Fig. 7

ZFIN ID: ZDB-FIG-180323-20

ZFIN ID: ZDB-FIG-180323-20

Herbert et al., 2017 - Dynein/dynactin is necessary for anterograde transport of Mbp mRNA in oligodendrocytes and for myelination in vivo.. Proceedings of the National Academy of Sciences of the United States of America 114:E9153-E9162 Full text @ Proc. Natl. Acad. Sci. USA

ADDITIONAL FIGURES

EXPRESSION / LABELING:

Gene:	mbpa
Fish:	WT
Condition:	chemical treatment by environment: ciliobrevin D
Anatomical Terms:	oligodendrocyte cell body oligodendrocyte glial cell projection
Stage:	Day 4

PHENOTYPE:

Fish:	WT
Condition:	chemical treatment by environment: ciliobrevin D
Observed In:	oligodendrocyte glial cell projection
Stage:	Day 4

Fig. 7

Acute dynein inhibition arrests both anterograde and retrograde Mbp mRNA transport in cultured oligodendrocytes and perturbs mbp localization in zebrafish. (A) In kymographs representing 60 s of live-cell imaging, MS2-GFP–labeled Mbp mRNA can be seen moving in the retrograde direction in untreated cells (Top). Following acute ciliobrevin D (15 μM) addition, Mbp mRNA displays bidirectional motility characterized by frequent back-and-forth movement and many directional switches (Middle). Finally, around 15 min after ciliobrevin D (15 μM) addition, most cells display arrested motility (Bottom). (B) A scatter plot represents the average anterograde or retrograde net speeds for individual oligodendrocytes. (C and D) Anterograde or retrograde net speeds were averaged for each cell and binned across 27-min time periods following ciliobrevin treatment. At 8–15 min following ciliobrevin treatment, speeds significantly decreased compared with earlier time points. One-way ANOVAs with post hoc Tukey’s test were performed. Anterograde speeds: No CilioD vs. 1–7 min, P = 0.11; No CilioD vs. 8–15 min, **P = 0.0011; 1–7 min vs. 8–15 min, P = 0.077. Retrograde speeds: No CilioD vs. 1–7 min, P = 0.67; No CilioD vs. 8–15 min, P = 0.11, 1–7 min vs. 8–15 min, *P = 0.027. (E) A model shows that Mbp mRNA granules can move processively in the anterograde and retrograde directions as well as bidirectionally and that each Mbp mRNA granule can simultaneously bind to both kinesin and the dynein/dynactin complex. (F–I) mbp ISH of zebrafish larvae treated with ciliobrevin D for 21 h. Larvae were scored as having strong (F), normal (G), reduced (H), or absent (I) mbp localization in oligodendrocyte processes (arrowheads, processes; arrows, cell bodies). (J) Quantification of phenotypic distribution shows that there was a significant difference in scores for DMSO (n = 71) and ciliobrevin D-treated larvae (n = 68), with the latter exhibiting more larvae with reduced or absent mbp localization in oligodendrocyte processes (P < 4.5E-14, Fisher’s exact test). (Scale bar, 50 μM.)

Gene Expression Details

Gene	Fish	Conditions	Stage	Anatomy	Assay
mbpa	WT	control	Day 4	oligodendrocyte cell body	ISH
			Day 4	oligodendrocyte glial cell projection	ISH
	WT	chemical treatment by environment: ciliobrevin D	Day 4	oligodendrocyte cell body	ISH
			Day 4	oligodendrocyte glial cell projection	ISH

Antibody Labeling Details No data available

Phenotype Details

Fish	Conditions	Stage	Phenotype
WT	chemical treatment by environment: ciliobrevin D	Day 4	oligodendrocyte glial cell projection mbpa expression decreased amount, abnormal

Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA