CRISPR-Cas9 Mediated TSPO Gene Knockout alters Respiration and Cellular Metabolism in Human Primary Microglia Cells

URN to cite this document:

urn:nbn:de:bvb:355-epub-405578

Milenkovic, Vladimir M. ; Slim, Dounia ; Bader, Stefanie ; Koch, Victoria ; Heinl, Elena-Sofia ; Alvarez-Carbonell, David ; Nothdurfter, Caroline ; Rupprecht, Rainer ; Wetzel, Christian H.

Preview

License: Creative Commons Attribution 4.0 PDF (2MB)

Date of publication of this fulltext: 23 Jul 2019 12:47

---

Alternative links to fulltext:DOIPublisher

---

Details

Item type:	Article
Date:	9 July 2019
Institutions:	Medicine > Lehrstuhl für Psychiatrie und Psychotherapie > Molekulare Neurowissenscahften
Projects:	Open Access Publizieren (DFG)
Identification Number:	Value Type 10.3390/ijms20133359 DOI
Keywords:	TSPO; mitochondria; knockdown; knockout; mitochondrial membrane potential; Ca2+ homeostasis; oxidative phosphorylation; steroid synthesis
Dewey Decimal Classification:	600 Technology > 610 Medical sciences Medicine
Status:	Published
Refereed:	Yes, this version has been refereed
Created at the University of Regensburg:	Yes
Item ID:	40557

Preview

Export bibliographical data

ASCII CitationAtomBibTeXData Cite XMLDataCite XML OAIDublin CoreEndNoteHTML CitationMETSMODSOpenAPCRDF+N-TriplesRDF+N3RDF+XMLReferReference ManagerSimple MetadataXMLxMetaDissPlus

---

Abstract

The 18 kDa translocator protein (TSPO) is an evolutionary conserved cholesterol binding protein localized in the outer mitochondrial membrane. It has been implicated in the regulation of various cellular processes including oxidative stress, proliferation, apoptosis, and steroid hormone biosynthesis. Since the expression of TSPO in activated microglia is upregulated in various neuroinflammatory ...

Abstract

The 18 kDa translocator protein (TSPO) is an evolutionary conserved cholesterol binding protein localized in the outer mitochondrial membrane. It has been implicated in the regulation of various cellular processes including oxidative stress, proliferation, apoptosis, and steroid hormone biosynthesis. Since the expression of TSPO in activated microglia is upregulated in various neuroinflammatory and neurodegenerative disorders, we set out to examine the role of TSPO in an immortalized human microglia C20 cell line. To this end, we performed a dual approach and used (i) lentiviral shRNA silencing to reduce TSPO expression, and (ii) the CRISPR/Cas9 technology to generate complete TSPO knockout microglia cell lines. Functional characterization of control and TSPO knockdown as well as knockout cells, revealed only low de novo steroidogenesis in C20 cells, which was not dependent on the level of TSPO expression or influenced by the treatment with TSPO-specific ligands. In contrast to TSPO knockdown C20 cells, which did not show altered mitochondrial function, the TSPO deficient knockout cells displayed a significantly decreased mitochondrial membrane potential and cytosolic Ca2+ levels, as well as reduced respiratory function. Performing the rescue experiment by lentiviral overexpression of TSPO in knockout cells, increased oxygen consumption and restored respiratory function. Our study provides further evidence for a significant role of TSPO in cellular and mitochondrial metabolism and demonstrates that different phenotypes of mitochondrial function are dependent on the level of TSPO expression.

---

---

Metadata last modified: 09 Aug 2019 11:12

Owner only: item control page

Download Statistics

Download Statistics

Download Statistics

Download Statistics

Downloads

Downloads per month over past year

Altmetric

Altmetric

Altmetric

Alternative Statistics (altmetrics)

More literature (via CORE)

More literature (via CORE)

More literature (via CORE)

More literature (via CORE)