Quantifying Compartment-Specific Protein Translocation in Astrocytes by Object-Oriented Image Analysis: Mitochondrial Translocation of PKC δ.

Quantifying Compartment-Specific Protein Translocation in Astrocytes by Object-Oriented Image Analysis: Mitochondrial Translocation of PKCδ. Methods Mol Biol. 2019;1938:169-186 Authors: Farid KMN, Derouiche A Abstract Assessing the amount and subcellular distribution of protein expression is a key component in modern cell biological and medical research. We studied protein kinase Cδ (PKCδ) as a potential regulator of mitochondrial metabolism in astrocytes, and sought to evaluate mitochondrial translocation of PKCδ since this is an important determinant of its function. Apart from visualizing compartment specific localization of mobile proteins such as PKCδ, we also wanted to determine what amount of a cell's total content of a particular protein is located to a specific compartment, or translocated comparing control and experimental condition.We develop a semiquantitative parameter that indicates the relative protein distribution to two subcellular compartments, starting from standard two-channel fluorescence microscopy images. We studied the mitochondrial translocation of PKCδ in astrocytes using double immunofluorescence microscopy and object-oriented image analysis. In one channel, the protein of interest (PKCδ) is labeled, in the other the compartment or organelle of interest (mitochondria, using cytochrome oxidase IV). Both channels were binarized, turned into object populations, and the channel specific values for total ...
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research