Weigelt, Karin and Ernst, Wolfgang and Walczak, Yana and Ebert, Stefanie and Loenhardt, Thomas and Klug, Maja and Rehli, Michael and Weber, Bernhard H. F. and Langmann, Thomas (2007) Dap12 expression in activated microglia from retinoschisin-deficient retina and its PU.1-dependent promoter regulation. JOURNAL OF LEUKOCYTE BIOLOGY, 82 (6). pp. 1564-1574. ISSN 0741-5400,
Full text not available from this repository. (Request a copy)Abstract
Several alterations in the expression of immune-related transcripts were identified recently in the degenerating retina of the retinoschisin knockout (Rs1h(-/Y)) mouse, including the strong expression of the adaptor protein Dap12. As Dap12 is found in leukocytes, we hypothesized that its disease-related expression may be confined to activated retinal microglia cells. To test this hypothesis, we established a procedure for isolation and culture of retinal microglia cells and performed genome-wide expression profiling from Rs1h(-/Y) and control microglia. While retaining their activated state in culture, ex vivo microglia expressed high levels of Dap12 and the transcription factor PU. 1. The activation-dependent induction of Dap12 was also confirmed in the microglia cell line BV-2 following in vitro stimulation. To examine the transcriptional regulation of Dap12 further, macrophage cell lines were transfected with several Dap12 reporter constructs. Promoter deletion assays and site-directed mutagenesis experiments demonstrated an essential role of evolutionarily conserved PU. 1 consensus sites in the proximal -104/+118 Dap12 promoter. In vitro and in vivo binding of PU. 1 to this promoter region was demonstrated using EMSA and chromatin immunoprecipitation. Knockdown of PU. 1 by RNA interference caused a significant reduction of endogenous Dap12 expression and re-expression, and activation of PU. 1 in PU. 1(-/-) progenitor cells induced Dap12 transcription. Taken together, our results indicate that activated microglia from degenerating retinae express high levels of Dap12 and PU. 1, and PU. 1 controls the myeloid-specific regulation of Dap12 directly and may also play a general role in microglia gene expression during retinal degeneration.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | STIMULATING FACTOR-RECEPTOR; GENE-EXPRESSION; MICROARRAY ANALYSIS; RAT RETINA; PU.1; MOUSE; TRANSCRIPTION; PROTEIN; CELLS; DIFFERENTIATION; Tyrobp; retinal degeneration; myeloid promoter |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Humangenetik Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 25 Nov 2020 09:32 |
| Last Modified: | 25 Nov 2020 09:32 |
| URI: | https://pred.uni-regensburg.de/id/eprint/31861 |
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