PUBLICATION
Cos2/Kif7 and OSM-3/Kif17 Regulate Onset of Outer Segment Development in Zebrafish Photoreceptors Through Distinct Mechanisms
- Authors
- Lewis, T.R., Kundinger, S.R., Pavlovich, A.L., Bostrom, J.R., Link, B.A., Besharse, J.C.
- ID
- ZDB-PUB-170328-10
- Date
- 2017
- Source
- Developmental Biology 425(2): 176-190 (Journal)
- Registered Authors
- Besharse, Joseph C., Lewis, Tylor, Link, Brian
- Keywords
- Hedgehog signaling, Intraflagellar Transport, Kinesin, Outer Segment
- MeSH Terms
-
- Animals
- Base Sequence
- CRISPR-Cas Systems/genetics
- Cilia/drug effects
- Cilia/metabolism
- Gene Editing
- Hedgehog Proteins/metabolism
- Kinesins/metabolism*
- Mice, Inbred C57BL
- Models, Biological
- Morphogenesis*/drug effects
- Morpholinos/pharmacology
- Mutation/genetics
- Neurogenesis/drug effects
- Retinal Photoreceptor Cell Outer Segment/metabolism*
- Signal Transduction/drug effects
- Temperature
- Transcription Activator-Like Effector Nucleases
- Zebrafish/metabolism*
- Zebrafish Proteins/metabolism*
- PubMed
- 28341548 Full text @ Dev. Biol.
Citation
Lewis, T.R., Kundinger, S.R., Pavlovich, A.L., Bostrom, J.R., Link, B.A., Besharse, J.C. (2017) Cos2/Kif7 and OSM-3/Kif17 Regulate Onset of Outer Segment Development in Zebrafish Photoreceptors Through Distinct Mechanisms. Developmental Biology. 425(2):176-190.
Abstract
Zebrafish morphants of osm-3/kif17, a kinesin-2 family member and intraflagellar transport motor, have photoreceptor outer segments that are dramatically reduced in number and size. However, two genetic mutant lines, osm-3/kif17sa0119 and osm-3/kif17sa18340, reportedly lack any observable morphological outer segment defects. In this work, we use TALENs to generate an independent allele, osm-3/kif17mw405, and show that both osm-3/kif17sa0119 and osm-3/kif17mw405 have an outer segment developmental delay in both size and density that is fully recovered by 6 days post-fertilization. Additionally, we use CRISPRs to generate cos2/kif7mw406, a mutation in the kinesin-4 family member cos2/kif7 that has been implicated in controlling ciliary architecture and Hedgehog signaling to test whether it may be functioning redundantly with osm-3/kif17. We show that cos2/kif7mw406 has an outer segment developmental delay similar to the osm-3/kif17 mutants. Using a three-dimensional mathematical model of outer segments, we show that while cos2/kif7mw406 and osm-3/kif17mw405 outer segments are smaller throughout the first 6 days of development, the volumetric rates of outer segment morphogenesis are not different among wild-type, cos2/kif7mw406, and osm-3/kif17mw405 after 60hpf. Instead, our model suggests that cos2/kif7mw406 and osm-3/kif17mw405 impact outer segment morphogenesis through upstream events that that are different for each motor. In the case of cos2/kif7mw406 mutants, we show that early defects in Hedgehog signaling lead to a general, non-photoreceptor-specific delay of retinal neurogenesis, which in turn causes the secondary phenotype of delayed outer segment morphogenesis. In contrast, the osm-3/kif17mw405 outer segment morphogenesis delays are linked specifically to initial disc morphogenesis of photoreceptors rather than an upstream event. Further, we show that osm-3/kif17 mutant mice also exhibit a similarly delayed outer segment development, suggesting a role for osm-3/kif17 in early outer segment development that is conserved across species. In conclusion, we show that both osm-3/kif17 and cos2/kif7 have comparable outer segment developmental delays, although through independent mechanisms.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping