Sura, Survasha; Department of Biological Sciences; Department of Biochemical and Molecular Biology; Georgia Cancer Center; Rajpurohit, Surendra K; Augusta University (2019-02-13)
      Zebrafish have emerged as a powerful model organism for elucidating the development and function of microglia. Generation of new transgenic reporter lines and imaging tools strengthen the zebrafish model in microglia study�in-vivo. The aim is to develop a novel compound transgenic line to study the inflammatory process mediated by NF-kB in microglia cells. This novel compound transgenic line will establish a new model for microglia study. To generate the novel compound zebrafish transgenic model for microglia, we are crossbreeding microglia transgenic line zebrafish (Tg(mpeg1:mCherry) with the NF-kB Tg(6xNFkB:EGFP) transgenic progeny. We first generate a heterozygous F1 progeny which will be bred to generate an F2 homozygous progeny. Once the F1 progeny of the Microglia-NfkB transgenic line is developed, they will be crossbred to develop the Homozygous compound transgenic line. Fluorescent Microscopy will be used to screen the larvae generated from the breeding events. By developing the compound transgenic line, we are optimizing microglia isolation and sorting methodology by using the related antibodies as the marker. The NF-kB microglia transgenic line will provide a unique platform for drug screening to address microglial based ailments, thus furthering the understanding and treatment of human disease.
    • Production of a NF-¿B Deficient Microglial Animal Model

      Goodall, Michael; Soni, Karan; Department of Biological Sciences; Bradford, Jennifer; Department of Biological Sciences; Augusta University (2018-02-12)
      Our goal is to determine how the nuclear factor-kappaB (NF-?B) signaling pathway is used in the communication between microglia and the progression of glioblastoma (GBM) cancer cells. The NF-?B signaling pathway is very important in normal immune system function and has been implicated in various types of cancers, including, GBM. GBM is the most common type of adult brain cancer, has altered NF-?B signaling, and is also characterized by a large population of microglia, the immune cell of the central nervous system. Based on our recent studies, we hypothesize that deleting the major transcription factor (p65) of the canonical NF-?B pathway in microglia would slow the progression of GBM. To test this hypothesis, we have developed a p65fl/fl/CX3CR1CreERtransgenic animal, which should lack microglial p65 after exposure to tamoxifen. We currently have heterozygous animals and will soon begin characterizing them to determine p65 deletion efficiency.