PUBLICATION

Localized TPC1-mediated Ca2+ release from endolysosomes contributes to myoseptal junction development in zebrafish

Authors
Rice, K.L., Webb, S.E., Miller, A.L.
ID
ZDB-PUB-220409-10
Date
2022
Source
Journal of Cell Science   135(9): (Journal)
Registered Authors
Miller, Andrew L., Webb, Sarah E.
Keywords
Ca2+ signaling, Endolysosomes, Myoseptal junctions, Skeletal slow muscle cells, Two-pore channel type 1, Zebrafish
MeSH Terms
  • Animals
  • Calcium*/metabolism
  • Dystroglycans/metabolism
  • Humans
  • Morpholinos
  • Muscle Fibers, Skeletal/metabolism
  • Muscle, Skeletal/metabolism
  • Zebrafish*/genetics
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
35393618 Full text @ J. Cell Sci.
Abstract
In the trunk of developing zebrafish embryos, adjacent myotome blocks transmit contractile force via myoseptal junctions (MJs), dynamic structures that connect the actin cytoskeleton of skeletal muscle cells to extracellular matrix components via transmembrane protein complexes in the sarcolemma. Here, we report that the endolysosomal ion channel, two-pore channel type 1 (TPC1), generates highly localized, non-propagating Ca2+ transients that play a distinct and required role in the capture and attachment of superficial slow skeletal muscle cells (SMCs) at MJs. Use of antisense morpholinos or CRISPR/Cas9 gene editing to disrupt tpcn1 gene expression resulted in abnormal MJ phenotypes including SMCs detaching from or crossing the myosepta. We also report that TPC1-decorated endolysosomes are dynamically associated with MJs in a microtubule-dependent manner, and that attenuating tpcn1 expression or function disrupted endolysosomal trafficking and resulted in an abnormal distribution of β-dystroglycan (a key transmembrane component of the dystrophin-associated protein complex). Together, our data suggest that localized TPC1-generated Ca2+ signals facilitate essential endolysosomal trafficking and membrane contact events, which help form and maintain MJs following the onset of SMC contractile activity.
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