Investigating microglia-brain tumor cell interactions in vivo in the larval zebrafish brain

Abstract

Glioblastoma is the most frequent and aggressive primary malignant brain tumor. Gliomas exhibit high genetic diversity in addition to complex and variable clinical features. Glioblastoma tumors are highly resistant to multimodal therapies and there is significant patient mortality within the first two years after prognosis. At present clinical treatments are palliative, not curative. Glioblastomas contain a high number of microglia and infiltrating macrophages, which are positively correlated with glioma grade and invasiveness. Microglia are the resident macrophages of the central nervous system. These cells constantly scan the brain and react promptly to any abnormality, removing detrimental factors and safeguarding the central nervous system against further damage. Microglia and macrophages that have colonized the glioblastoma display protumoral functions and promote tumor growth. The optically transparent zebrafish larva facilitates imaging of fluorescently labeled cells at high spatial and temporal resolution in vivo. It is therefore an excellent model to investigate microglia-glioma cell interactions at the early stages of tumor development. Here we provide several methods that can be used to study the early stages of microglia-glioma cell interactions in the zebrafish. We present a technique for the xenotransplantation of mammalian oncogenic cells into the zebrafish brain and provide advice for image capture and analysis.

Keywords: Glioblastoma; Live-imaging; Microglia; Xenotransplantation; Zebrafish.