Quantifying inter-organelle membrane contact sites using proximity ligation assay in fixed optic nerve sections.

ABSTRACT: Membrane contact sites (MCS) play crucial roles in cell physiology with dysfunction in several MCS proteins being linked with neurological and optic nerve diseases. Although there have been significant advances in imaging these interactions over the past two decades with advanced electron microscopy techniques, super-resolution imaging and proximity-dependent fluorescent reporters, a technique to observe and quantify MCS in mammalian optic nerve tissues has not yet been reported. We demonstrate for the first time that proximity ligation assay (PLA), a technique already used in mammalian cell lines, can be used as an efficient method of quantifying inter-organelle contact sites, namely mitochondria-endoplasmic reticulum (ER) and mitochondria-late-endosomes, in mammalian optic nerve tissues

SUBMITTER: Ching J 

PROVIDER: S-EPMC8683748 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature