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. 2022 Nov 8;16:1047928. doi: 10.3389/fncel.2022.1047928

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Copyright © 2022 Smith, Park, Aalderink, Oldfield, Bergin, Mee, Faull and Dragunow.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Basal and Macrophage Colony-Stimulating Factor (M-CSF)-induced microglial proliferation and expression of PU.1, CSF-1R, DAP12, CEBPβ, and IGF1 receptor. (A) Quantification of the percentage of microglia that incorporate BrdU under control conditions and with M-CSF treatment showing a significant increase in microglial division with M-CSF and an enhanced response in ventricular/Hp microglia. Each data point indicates an individual case (n = 7). (B) M-CSF significantly increases the intensity (arbitrary fluorescence units) of PU.1 expression (amount of PU.1 protein) in adult human microglia from neurogenic (ventricular/Hp) and non-neurogenic (cortical) regions of the adult human brain. (C) CSF-1R is more highly expressed in ventricular/Hp microglia than cortical microglia. A significant increase in intensity of receptor labelling is found for CSF-1R following M-CSF treatment. (D) Quantification of DAP12 staining intensity shows a significantly greater level of expression in ventricular/Hp microglia compared to cortical microglia, and a significant increase in DAP12 expression with M-CSF treatment. (E) Quantification of microglial C/EBPβ expression showing differential basal expression by cortical and ventricular/Hp microglia, and significant increases in the percentage of cortical and ventricular/Hp microglia which express C/EBPβ following M-CSF treatment. (F) Ventricular/Hp microglia express significantly more IGF-1R than cortical microglia, and a significant increase in intensity of IGF-1R is evident following M-CSF treatment. (B–F) Protein expression was quantified using an intensity threshold in Metamorph image analysis software, as described in the methods. *indicates P value < 0.05.