TY  - JOUR
TI  - Preparation of Caenorhabditis elegans for Scoring of Muscle-derived Exophers
AU  - Banasiak, Katarzyna 
AU  - Turek, Michał 
AU  - Pokrzywa, Wojciech 
VL  - 13
IS  - 1
PY  - 2023
DA  - 2023/01/05
SP  - e4586
C1  - Bio-protocol 2023;13:e4586
DO  - 10.21769/BioProtoc.4586
UR  - https://doi.org/10.21769/BioProtoc.4586
AB  - Utilizingresources available from the mother's body to guarantee healthy offspring growth is the fundamental reproductive strategy. Recently, we showed that a class of the largest extracellular vesicles known as exophers, which are responsible for the removal of neurotoxic components from neurons (Melentijevic et al., 2017) and damaged mitochondria from cardiomyocytes (Nicolás-Ávila et al., 2020), are released by the Caenorhabditis elegans hermaphrodite body wall muscles (BWM), to support embryonic growth (Turek et al., 2021). Employing worms expressing fluorescent reporters in BWM cells, we found that exopher formation (exophergenesis) is sex-specific and fertility-dependent. Moreover, exophergenesis is regulated by the developing embryo in utero, and exophers serve as transporters for muscle-generated yolk proteins, which can be used to nourish the next generation. Given the specific regulation of muscular exophergenesis, and the fact that muscle-generated exophers are much larger than neuronal ones and have different targeting, their identification and quantification required a modified approach from that designed for neuronal-derived exophers (Arnold et al., 2020). Here, we present a methodology for assessing and quantifying muscle-derived exophers that can be easily extended to determine their function and regulation in various biological contexts.							Graphical abstract
KW  - C. elegans
KW  - Body wall muscle
KW  - Extracellular vesicle
KW  - Exopher
KW  - Fluorescent microscopy
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False