Characterisation of microglia-specific Tgfbr2, Smad4 and Tgfb1 knockout mice

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Zusammenfassung: This thesis was conducted to describe the role of TGFβ mediated regulation of microglia homeostasis and activation. To this aim, I generated and characterised microglia-specific knockout mouse lines to analyse the importance of R-SMAD-dependent (mgTgfbr2-/-) and Co-SMAD-dependent (mgSmad4-/-) TGFβ signalling in microglia. Moreover, microglia-specific deletion of Tgfb1 (mgTgfb1-/-) was induced to address the effects of microglia-derived TGFβ1. According to the current literature, this is the first study using mouse lines with microglia-specific knockout of TGFβ signalling molecules.I could show that the generated mouse lines are valid models for the microglia-specific analysis of TGFβ signalling, as demonstrated by deletion on genomic and protein levels. Importantly, functional impairment of TGFβ signalling could be detected in vitro and in vivo. The knockout mice were used to reveal that microglial TGFβ signalling is not essential for microglia survival and maintenance, as no differences were seen in microglia cell numbers in vivo. Elevated cell numbers in vitro indicated an increased proliferative activity of mgTgfbr2-/- microglia. Furthermore, mice did not show any phenotypic differences with regard to body weight or survival. Additionally, using mgSmad4-/- mice, I could show that the expression of distinct TGFβ-induced genes was dependent on SMAD4. Interestingly, TGFβ-driven nuclear translocation of R-SMADs in microglia was independent on SMAD4.I could show that knockout of microglial Tgfbr2, but not of Smad4 or Tgfb1, led to an activated microglia phenotype under physiological conditions as reflected by increased expression activation markers CD86, CD206, CD36 and MHCII. mgTgfbr2-/- microglia also showed an increase of TAK1 phosphorylation as well as CCL2 and CXCL10 secretion. In contrast, knockout of Smad4 only led to a moderate increase of TAK1 phosphorylation and secretion of CCL3 and CCL4 in vitro. I could therefore show that the effects mgTgfbr2 and mgSmad4 knockout resulted in different outcome, indicating a major role of R-SMAD-dependent signalling in the regulation of microglia activation under physiological conditions. However, distinct and partially non-overlapping effects were detected in mgSmad4-/- microglia, indicating a contribution of Co-SMAD-dependent signalling for microglia quiescence. Exogenous TGFβ1 was no longer able to attenuate IFNγ-induced upregulation of the activation markers iNos and Tnfa in Tgfbr2- and Smad4-deficient microglia, further underlining the importance of TGFβ-signalling to regulate microglia activation. Next, I could show that microglia-derived TGFβ1 is not essential for microglia quiescence in vitro and in vivo, as mgTgfb1-/- mice did not display the activated phenotype observed in mgTgfbr2-/- mice, as well as in Tgfb1 knockout mice described in recent studies. In this thesis, microglia-specific knockouts of Tgfbr2, Smad4 and Tgfb1 were generated and characterised for the first time. These mice will serve as valuable tools in upcoming projects addressing the role of microglial TGFβ signalling in models of injury and neurodegeneration and will further increase our understanding of TGFβ-mediated effects on microglia during neurodegenerative diseases

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch
• Notes: Albert-Ludwigs-Universität Freiburg, Dissertation, 2016

Classification and Topics

• Keyword: Hirnkrankheit ; Neuropsychiatrie ; Nervenkrankheit

Contributors, Places and Time

• Event: Veröffentlichung
• (where): Freiburg
• (who): Universität
• (when): 2016
• Creator: Zöller, Tanja
• Contributor: Spittau, Björn

Further information

• DOI: 10.6094/UNIFR/10889
• URN: urn:nbn:de:bsz:25-freidok-108894
• Rights: Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 25.03.2025, 1:42 PM CET