The Impact of Fixation on the Detection of Oligodendrocyte Precursor Cell Morphology and Vascular Associations.

Friederike Pfeiffer,Akiko Nishiyama,Amin Sherafat

doi:10.3390/cells10061302

Friederike Pfeiffer, Akiko Nishiyama + Show 1 more

Open Access

https://doi.org/10.3390/cells10061302

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Journal: Cells	Publication Date: May 24, 2021
Citations: 7	License type: CC BY 4.0

Affiliation: University of Connecticut, University of Tübingen

Abstract

Oligodendrocyte precursor cells (OPCs) display numerous protrusions that extend into the surrounding parenchyma in the brain. Depending on the preparation of the tissue analyzed, these protrusions are more or less visible. We applied six different fixation methods and compared the effect of prolonged and stronger fixation on fluorescence intensity of platelet-derived growth factor receptor alpha, a surface marker of OPCs. Importantly, the fluorescence signal is mostly lost on protrusions as compared to the cell body, which has to be considered for specific analyses. Additionally, we show numerous contacts established between OPCs and the brain vasculature, which will contribute to the understanding of the interactions between these two elements.

Highlights

Oligodendrocyte precursor cells (OPCs) cells comprise a glial cell population that covers the entire parenchyma of the central nervous system (CNS)
Investigations of the functions of OPCs require a detailed assessment of OPC morphology and localization, at all the sites of contact they establish with other cells of the CNS both by light and electron microscopy
To achieve better detection of the cell surface markers for OPCs, we have been routinely using 4% PFA containing 0.1 M L-lysine and 0.01 M sodium metaperiodate (PLP), which is suitable for detecting cell surface glycoprotein antigens, including NG2 [9] and PDGFRα [10]

Summary

Introduction

Oligodendrocyte precursor cells (OPCs) cells comprise a glial cell population that covers the entire parenchyma of the central nervous system (CNS). In addition to generating oligodendrocytes, some OPCs do not differentiate for months but remain as OPCs [1,2] These non-vascular OPCs express the NG2 antigen and the alpha receptor for platelet-derived growth factor (PDGFRα), both of which are localized on the cell surface [3] and are commonly used to detect and visualize OPCs in immunohistochemical studies by antibodies against those proteins. Investigations of the functions of OPCs require a detailed assessment of OPC morphology and localization, at all the sites of contact they establish with other cells of the CNS both by light and electron microscopy. To this end, it is absolutely essential to optimize methods to visualize their numerous processes accurately and consistently at a high resolution. The quality of immunodetection largely depends on how the tissue is prepared

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