PUBLICATION

In vivo cell biological screening identifies an endocytic capture mechanism for T-tubule formation

Authors
Hall, T.E., Martel, N., Ariotti, N., Xiong, Z., Lo, H.P., Ferguson, C., Rae, J., Lim, Y.W., Parton, R.G.
ID
ZDB-PUB-200728-18
Date
2020
Source
Nature communications   11: 3711 (Journal)
Registered Authors
Hall, Thomas, Lo, Harriet, Parton, Robert G.
Keywords
none
MeSH Terms
  • Zebrafish
  • Myofibrils/metabolism
  • Golgi Apparatus/metabolism
  • Calcium Channels/metabolism
  • Calcium Channels/ultrastructure
  • Developmental Biology
  • Calcium Channels, L-Type/metabolism
  • Male
  • Microscopy, Electron
  • Sarcoplasmic Reticulum/metabolism
  • Muscle Proteins/chemistry
  • Muscle Proteins/metabolism*
  • Muscle Contraction/physiology*
  • Sarcolemma/chemistry
  • Sarcolemma/physiology*
  • Sarcolemma/ultrastructure*
  • Muscle, Skeletal/chemistry
  • Muscle, Skeletal/metabolism*
  • Muscle, Skeletal/ultrastructure*
  • Carrier Proteins/metabolism
  • Animals
(all 21)
PubMed
32709891 Full text @ Nat. Commun.
Abstract
The skeletal muscle T-tubule is a specialized membrane domain essential for coordinated muscle contraction. However, in the absence of genetically tractable systems the mechanisms involved in T-tubule formation are unknown. Here, we use the optically transparent and genetically tractable zebrafish system to probe T-tubule development in vivo. By combining live imaging of transgenic markers with three-dimensional electron microscopy, we derive a four-dimensional quantitative model for T-tubule formation. To elucidate the mechanisms involved in T-tubule formation in vivo, we develop a quantitative screen for proteins that associate with and modulate early T-tubule formation, including an overexpression screen of the entire zebrafish Rab protein family. We propose an endocytic capture model involving firstly, formation of dynamic endocytic tubules at transient nucleation sites on the sarcolemma, secondly, stabilization by myofibrils/sarcoplasmic reticulum and finally, delivery of membrane from the recycling endosome and Golgi complex.
Genes / Markers
Figures
Figure Gallery (9 images)
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
pc10TgTransgenic Insertion
    pc21TgTransgenic Insertion
      pc22TgTransgenic Insertion
        uq4rpTgTransgenic Insertion
          uq20rp
            Small Deletion
            uq21rp
              Indel
              uq22rpTgTransgenic Insertion
                1 - 7 of 7
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                Human Disease / Model
                No data available
                Sequence Targeting Reagents
                Target Reagent Reagent Type
                smyhc1CRISPR5-smyhc1CRISPR
                ttn.2CRISPR16-ttn.2CRISPR
                tyrCRISPR1-tyrCRISPR
                1 - 3 of 3
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                Fish
                Antibodies
                No data available
                Orthology
                No data available
                Engineered Foreign Genes
                Marker Marker Type Name
                APXEFGAPX
                EGFPEFGEGFP
                mCherryEFGmCherry
                mKate2EFGmKate2
                1 - 4 of 4
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                Mapping
                No data available