Related Experiment Video
Updated: Dec 29, 2025
Optogenetic Phase Transition of TDP-43 in Spinal Motor Neurons of Zebrafish Larvae
Published on: February 25, 2022
Temporal and spatial expression of zebrafish mctp genes and evaluation of frameshift alleles of mctp2b
Angeles E Espino-Saldaña1, Karina Durán-Ríos2, Eduardo Olivares-Hernandez2
1Instituto de Neurobiología, Universidad Nacional Autónoma de México, Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Juriquilla, Querétaro, 76230, Mexico; Universidad Autónoma de Querétaro, Facultad de Ciencias Naturales, Av. de las Ciencias S/N, Querétaro, Mexico.
Abstract:
MCTPs (multiple C2 domain proteins with two transmembrane regions) have been proposed as novel endoplasmic reticulum calcium sensors; however, their function remains largely unknown. Here we report the structure of the four mctp genes from zebrafish (mctp1a, mctp1b, mctp2a and mctp2b), their diversity, expression pattern during embryonic development and in adult tissue and the effect of knocking down the expression of Mctp2b by CRISPR/Cas9. The four mctp genes are expressed from early development and exhibit differential expression patterns but are found mainly in the nervous and muscular systems. Mctp2b tagged with fluorescent proteins and expressed in HEK-293 cells and neurons of the fish spinal cord localized mostly in the endoplasmic reticulum but also in lysosomes and late and recycling endosomes. Knocking down mctp2b expression impaired embryonic development, suggesting that the functional participation of this gene is relevant, at least during the early stages of development.
Insights
Multiple C2 domain proteins (MCTPs) are novel endoplasmic reticulum calcium sensors. Zebrafish MCTP gene expression and Mctp2b knockdown reveal their crucial role in embryonic development, particularly in nervous and muscular systems.
Area of Science:
- Cell Biology
- Developmental Biology
- Genomics
Background:
- Multiple C2 domain proteins (MCTPs) are implicated as endoplasmic reticulum calcium sensors.
- The precise functions of MCTPs remain largely uncharacterized.
- Understanding MCTP roles is crucial for cellular calcium signaling research.
Purpose of the Study:
- To investigate the structure and diversity of four zebrafish MCTP genes (mctp1a, mctp1b, mctp2a, mctp2b).
- To analyze the expression patterns of MCTP genes during zebrafish embryonic development and in adult tissues.
- To determine the functional impact of Mctp2b knockdown on zebrafish development using CRISPR/Cas9 technology.
Main Methods:
- Gene sequencing and structural analysis of four zebrafish mctp genes.
- Quantitative expression analysis during embryonic development and in adult tissues.
- CRISPR/Cas9-mediated knockdown of mctp2b and subsequent phenotypic analysis.
- Fluorescent protein tagging and subcellular localization studies in HEK-293 cells and zebrafish neurons.
Main Results:
- Four zebrafish mctp genes were identified, exhibiting diverse expression patterns, primarily in nervous and muscular systems.
- Mctp2b localizes to the endoplasmic reticulum, lysosomes, and endosomes.
- Knockdown of mctp2b significantly impaired zebrafish embryonic development.
Conclusions:
- Zebrafish MCTP genes are expressed early in development with specific tissue distribution.
- Mctp2b plays a significant role in embryonic development, likely through its functions in the endoplasmic reticulum and endosomal pathways.
- Further research into MCTPs as calcium sensors and their developmental roles is warranted.
