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Global Deletion of TSPO Does Not Affect the Viability and Gene Expression Profile.

ABSTRACT: Translocator Protein (18kDa, TSPO) is a mitochondria outer membrane transmembrane protein and which expression is elevated during inflammation and injury. However, the exact function of TSPO is still controversial. Here, we constructed TSPO global knockout (KO) mice by Cre-LoxP system independently, which abolished TSPO protein expression in all tissues and shown normal phenotypes. The birth rate of TSPO heterozygote (Het)/Het breeding was consistent with the Mendel’s Law, suggesting a normal viability of TSPO KO mice at birth. RNAseq analysis showed no significant difference in the gene expression profile of lung tissues from TSPO KO mice compared with that of wild type mice, even the genes associated with bronchial alveoli immune homeostasis. The alveolar macrophage population was also not affected by TSPO deletion in the physiological condition. Our findings contradicts the results of Papadopoulos, but confirmed Selvaraj’s findings. This study reveals TSPO deficiency does not affect the viability and bronchial alveoli immune homeostasis.

ORGANISM(S): Mus musculus

PROVIDER: GSE84942 | GEO | 2016/12/08

SECONDARY ACCESSION(S): PRJNA335721

REPOSITORIES: GEO

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Project description:We hypothesized that the potential ability of the 18 kDa mitochondrial translocator protein (TSPO) to regulate gene expression may underlie its numerous reported functions, ranging from mitochondrial activity till mental disorders. Therefore, we applied microarray analysis of gene expression to U118MG glioblastoma cells. Within 1 hr the TSPO ligand PK 11195 induced changes in expression of immediate early genes and transcription factors. After 24 hrs primarily gene expression for enzymes and other proteins is changed. The associated gene products are known to affect various cellular functions: programmed cell death, cell cycle, viability, proliferation, migration, differentiation, development, tumorigenesis, and inflammatory / immune responses. Our microscopic studies showed intense TSPO mitochondrial labeling but no TSPO signal in the cell nuclei. Thus, it appears that the TSPO is part of the retrograde mitochondrial-nuclear signaling pathway for modulation of gene expression and thereby may exert its numerous effects on cell function, phenotype, and disease. U118MG cells (1.255 Ã 106) were seeded in Petri dishes (i.e. 28.5 Ã 103 cells / cm2) and allowed to proliferate for 3 days in full medium. Experiments for gene expression assayed with microarray typically consisted of three experimental groups and a vehicle control group (n = 3 for each group). Serum deprived medium was applied for 24 hrs, in the experimental groups ending with the inclusion a choice of various PK 11195 exposures i.e. either 1 hr, 3 hrs, or 24 hrs. Exposure to PK 11195 implies serum deprived medium with 1% alcohol (vehicle) and PK 11195 (25 ÂµM final concentration) for the required time period. 25 ÂµM of PK 11195 is the optimal concentration for these types of experiments with U118MG cells (Kugler et al., 2008). The cells were collected by trypsinization, washed by centrifugation in phosphate buffered saline (PBS) (400 Ã g, 5 min), and lysed [RLT lysis buffer provided with the RNeasy Mini Kit diluted with Î²-mercaptoethanol (1 : 100)], according to the manufacturer's instructions. Lysates were stored at -70oC.

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Global Deletion of TSPO Does Not Affect the Viability and Gene Expression Profile.

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