Collar occupancy: A new quantitative imaging tool for morphometric analysis of oligodendrocytes

• Type: Article in International Scientific Journal
• Date: 2018

Authors

Bouçanova, F (Author) Other; Maia, AF (Author) Other; Cruz, A (Author) Other; Millar, V (Author) Other; Mendes Pinto, I (Author) Other; Relvas, JB (Author) ICBAS; Domingues, HS (Author) Other

Journal

Title: Journal of Neuroscience Methods

Vol. 294

Pages: 122-135

ISSN: 0165-0270

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 0 Citations

Scopus - 3 Citations

Other information

Authenticus ID: P-00T-469

DOI: 10.1016/j.jneumeth.2017.11.014

Abstract (EN): Background: Oligodendrocytes (OL) are the myelinating cells of the central nervous system. OL differentiation from oligodendrocyte progenitor cells (OPC) is accompanied by characteristic stereotypical morphological changes. Quantitative imaging of those morphological alterations during OPC differentiation is commonly used for characterization of new molecules in cell differentiation and myelination and screening of new pro-myelinating drugs. Current available imaging analysis methods imply a non automated morphology assessment, which is time-consuming and prone to user subjective evaluation. New method: Here, we describe an automated high-throughput quantitative image analysis method entitled collar occupancy that allows morphometric ranking of different stages of in vitro OL differentiation in a high-content analysis format. Collar occupancy is based on the determination of the percentage of area occupied by OPC/OL cytoplasmic protrusions within a defined region that contains the protrusion network, the collar. Results: We observed that more differentiated cells have higher collar occupancy and, therefore, this parameter correlates with the degree of OL differentiation. Comparison with existing methods: In comparison with the method of manual categorization, we found the collar occupancy to be more robust and unbiased. Moreover, when coupled with myelin basic protein (MBP) staining to quantify the percentage of myelinating cells, we were able to evaluate the role of new molecules in OL differentiation and myelination, such as Dusp19 and Kank2. Conclusions: Altogether, we have successfully developed an automated and quantitative method to morphologically characterize OL differentiation in vitro that can be used in multiple studies of OL biology.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 14