PUBLICATION

Zebrafish raptor mutation inhibits the activity of mTORC1, inducing craniofacial defects due to autophagy-induced neural crest cell death

Authors
Tucker, S.K., Gosul, R., Swartz, M.E., Zhang, S., Eberhart, J.K.
ID
ZDB-PUB-240322-2
Date
2024
Source
Development (Cambridge, England)   151(6): (Journal)
Registered Authors
Eberhart, Johann, Swartz, Mary
Keywords
Autophagy, Cell death, Craniofacial, MTOR, Neural crest, Zebrafish
MeSH Terms
  • Mechanistic Target of Rapamycin Complex 1/metabolism
  • Animals
  • Signal Transduction/genetics
  • Cell Death
  • Regulatory-Associated Protein of mTOR/genetics
  • Regulatory-Associated Protein of mTOR/metabolism
  • TOR Serine-Threonine Kinases/genetics
  • TOR Serine-Threonine Kinases/metabolism
  • Zebrafish*/genetics
  • Zebrafish*/metabolism
  • Autophagy/genetics
  • Neural Crest*/metabolism
  • Mutation/genetics
(all 13)
PubMed
38512806 Full text @ Development
Abstract
The mechanistic target of rapamycin (mTOR) coordinates metabolism and cell growth with environmental inputs. mTOR forms two functional complexes: mTORC1 and mTORC2. Proper development requires both complexes but mTORC1 has unique roles in numerous cellular processes, including cell growth, survival and autophagy. Here, we investigate the function of mTORC1 in craniofacial development. We created a zebrafish raptor mutant via CRISPR/Cas9, to specifically disrupt mTORC1. The entire craniofacial skeleton and eyes were reduced in size in mutants; however, overall body length and developmental timing were not affected. The craniofacial phenotype associates with decreased chondrocyte size and increased neural crest cell death. We found that autophagy is elevated in raptor mutants. Chemical inhibition of autophagy reduced cell death and improved craniofacial phenotypes in raptor mutants. Genetic inhibition of autophagy, via mutation of the autophagy gene atg7, improved facial phenotypes in atg7;raptor double mutants, relative to raptor single mutants. We conclude that finely regulated levels of autophagy, via mTORC1, are crucial for craniofacial development.
Errata / Notes
This article is corrected by ZDB-PUB-240429-14.
Genes / Markers
Figures
Figure Gallery (12 images) / 2
Show all Figures
Expression
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay Figure
rptorAb1-rptorABstandard conditionsDay 4WB
rptorAb1-rptorrptorau93/au93 (AB)standard conditionsDay 4WB
rptory1Tg (AB)standard conditionsLong-pecISH
1 - 3 of 3
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Phenotype
Fish Conditions Stage Phenotype Figure
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768 (AB)standard conditionsDay 4
atg7sa14768/sa14768; rptorau93/au93 (AB)standard conditionsDay 4
atg7sa14768/sa14768; rptorau93/au93 (AB)standard conditionsDay 4
1 - 10 of 59
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
au93
    		Small Deletion
    sa14768
      		Point Mutation
      y1TgTransgenic Insertion
        zf155TgTransgenic Insertion
          1 - 4 of 4
          Show
          Human Disease / Model
          No data available
          Sequence Targeting Reagents
          Target Reagent Reagent Type
          rptorCRISPR2-rptorCRISPR
          1 - 1 of 1
          Show
          Fish
          Fish
          AB
          atg7sa14768/sa14768 (AB)
          atg7sa14768/sa14768; rptorau93/au93 (AB)
          rptorau93/au93 (AB)
          rptorau93/au93; y1Tg (AB)
          rptorau93/au93; zf155Tg (AB)
          y1Tg (AB)
          1 - 7 of 7
          Show
          Antibodies
          Name Type Antigen Genes Isotypes Host Organism
          Ab1-rptormonoclonalRabbit
          Ab2-eif4ebp1monoclonalIgGRabbit
          Ab5-rps6monoclonal
            		IgGRabbit
            Ab6-map1lc3bpolyclonal
              		IgGRabbit
              Ab7-rps6kmonoclonal
                		Rabbit
                Ab14-rps6kmonoclonal
                  		IgGRabbit
                  1 - 6 of 6
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                  Orthology
                  No data available
                  Engineered Foreign Genes
                  Marker Marker Type Name
                  EGFPEFGEGFP
                  1 - 1 of 1
                  Show
                  Mapping
                  No data available
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                  Citation
                  Tucker, S.K., Gosul, R., Swartz, M.E., Zhang, S., Eberhart, J.K. (2024) Zebrafish raptor mutation inhibits the activity of mTORC1, inducing craniofacial defects due to autophagy-induced neural crest cell death. Development (Cambridge, England). 151(6):.