PUBLICATION

Ca²⁺ release via two-pore channel type 2 (TPC2) is required for slow muscle cell myofibrillogenesis and myotomal patterning in intact zebrafish embryos.

Authors

Kelu, J.J., Webb, S.E., Parrington, J., Galione, A., Miller, A.L.

ID

ZDB-PUB-170410-9

Date

2017

Source

Developmental Biology 425(2): 109-129 (Journal)

Registered Authors

Galione, Antony, Miller, Andrew L., Webb, Sarah E.

Keywords

CRISPR/Cas9, Ca(2+) signaling, Morpholino oligonucleotides, Myofibrillogenesis, STED super-resolution microscopy, Slow skeletal muscle cell differentiation, Two-pore channel 2, Zebrafish

MeSH Terms

Animals
Base Sequence
Behavior, Animal/drug effects
Body Patterning*/drug effects
CRISPR-Cas Systems/genetics
Caffeine/pharmacology
Calcium/metabolism*
Calcium Channels/metabolism*
Calcium Signaling/drug effects
Cell Death/drug effects
Cells, Cultured
Embryo, Nonmammalian/drug effects
Embryo, Nonmammalian/metabolism*
Gene Knockdown Techniques
Gene Knockout Techniques
Inositol 1,4,5-Trisphosphate Receptors/metabolism
Kinesins/metabolism*
Macrolides/pharmacology
Models, Biological
Morpholinos/pharmacology
Motor Activity/drug effects
Muscle Cells/cytology
Muscle Cells/drug effects
Muscle Cells/metabolism
Muscle Development*/drug effects
Muscle Fibers, Slow-Twitch/cytology
Muscle Fibers, Slow-Twitch/drug effects
Muscle Fibers, Slow-Twitch/metabolism*
Phenotype
RNA, Messenger/genetics
RNA, Messenger/metabolism
Ryanodine Receptor Calcium Release Channel/metabolism
Sarcomeres/drug effects
Sarcomeres/metabolism
Zebrafish/embryology*
Zebrafish/metabolism*
Zebrafish Proteins/metabolism*

PubMed

28390800 Full text @ Dev. Biol.

Abstract

We recently demonstrated a critical role for two-pore channel type 2 (TPC2)-mediated Ca²⁺ release during the differentiation of slow (skeletal) muscle cells (SMC) in intact zebrafish embryos, via the introduction of a translational-blocking morpholino antisense oligonucleotide (MO). Here, we extend our study and demonstrate that knockdown of TPC2 with a non-overlapping splice-blocking MO, knockout of TPC2 (via the generation of a tpcn2^dhkz1a mutant line of zebrafish using CRISPR/Cas9 gene-editing), or the pharmacological inhibition of TPC2 action with bafilomycin A1 or trans-ned-19, also lead to a significant attenuation of SMC differentiation, characterized by a disruption of SMC myofibrillogenesis and gross morphological changes in the trunk musculature. When the morphants were injected with tpcn2-mRNA or were treated with IP₃/BM or caffeine (agonists of the inositol 1,4,5-trisphosphate receptor (IP₃R) and ryanodine receptor (RyR), respectively), many aspects of myofibrillogenesis and myotomal patterning (and in the case of the pharmacological treatments, the Ca²⁺ signals generated in the SMCs), were rescued. STED super-resolution microscopy revealed a close physical relationship between clusters of RyR in the terminal cisternae of the sarcoplasmic reticulum (SR), and TPC2 in lysosomes, with a mean estimated separation of ~52-87nm. Our data therefore add to the increasing body of evidence, which indicate that localized Ca²⁺ release via TPC2 might trigger the generation of more global Ca²⁺ release from the SR via Ca²⁺-induced Ca²⁺ release.

Genes / Markers

Figures

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Expression

Phenotype

Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Fish

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Kelu et al., 2017 ZDB-PUB-170410-9 PMID:28390800

Ca2+ release via two-pore channel type 2 (TPC2) is required for slow muscle cell myofibrillogenesis and myotomal patterning in intact zebrafish embryos.

Kelu et al., 2017

ZDB-PUB-170410-9
PMID:28390800

Ca²⁺ release via two-pore channel type 2 (TPC2) is required for slow muscle cell myofibrillogenesis and myotomal patterning in intact zebrafish embryos.