Cre-mediated stress affects sirtuin expression levels, peroxisome biogenesis and metabolism, antioxidant and proinflammatory signaling pathways.

TY - JOUR

T1 - Cre-mediated stress affects sirtuin expression levels, peroxisome biogenesis and metabolism, antioxidant and proinflammatory signaling pathways.

AU - Xiao, Yu

AU - Karnati, Srikanth

AU - Qian, Guofeng

AU - Nenicu, Anca

AU - Fan, Wei

AU - Tchatalbachev, Svetlin

AU - Höland, Anita

AU - Hossain, Hamid

AU - Guillou, Florian

AU - Lüers, Georg

AU - Baumgart-Vogt, Eveline

PY - 2012

Y1 - 2012

N2 - Cre-mediated excision of loxP sites is widely used in mice to manipulate gene function in a tissue-specific manner. To analyze phenotypic alterations related to Cre-expression, we have used AMH-Cre-transgenic mice as a model system. Different Cre expression levels were obtained by investigation of C57BL/6J wild type as well as heterozygous and homozygous AMH-Cre-mice. Our results indicate that Cre-expression itself in Sertoli cells already has led to oxidative stress and lipid peroxidation (4-HNE lysine adducts), inducing PPAR?/?, peroxisome proliferation and alterations of peroxisome biogenesis (PEX5, PEX13 and PEX14) as well as metabolic proteins (ABCD1, ABCD3, MFP1, thiolase B, catalase). In addition to the strong catalase increase, a NRF2- and FOXO3-mediated antioxidative response (HMOX1 of the endoplasmic reticulum and mitochondrial SOD2) and a NF-?B activation were noted. TGF?1 and proinflammatory cytokines like IL1, IL6 and TNF? were upregulated and stress-related signaling pathways were induced. Sertoli cell mRNA-microarray analysis revealed an increase of TNFR2-signaling components. 53BP1 recruitment and expression levels for DNA repair genes as well as for p53 were elevated and the ones for related sirtuin deacetylases affected (SIRT 1, 3-7) in Sertoli cells. Under chronic Cre-mediated DNA damage conditions a strong downregulation of Sirt1 was observed, suggesting that the decrease of this important coordinator between DNA repair and metabolic signaling might induce the repression release of major transcription factors regulating metabolic and cytokine-mediated stress pathways. Indeed, caspase-3 was activated and increased germ cell apoptosis was observed, suggesting paracrine effects. In conclusion, the observed wide stress-induced effects and metabolic alterations suggest that it is essential to use the correct control animals (Cre/Wt) with matched Cre expression levels to differentiate between Cre-mediated and specific gene-knock out-mediated effects.

AB - Cre-mediated excision of loxP sites is widely used in mice to manipulate gene function in a tissue-specific manner. To analyze phenotypic alterations related to Cre-expression, we have used AMH-Cre-transgenic mice as a model system. Different Cre expression levels were obtained by investigation of C57BL/6J wild type as well as heterozygous and homozygous AMH-Cre-mice. Our results indicate that Cre-expression itself in Sertoli cells already has led to oxidative stress and lipid peroxidation (4-HNE lysine adducts), inducing PPAR?/?, peroxisome proliferation and alterations of peroxisome biogenesis (PEX5, PEX13 and PEX14) as well as metabolic proteins (ABCD1, ABCD3, MFP1, thiolase B, catalase). In addition to the strong catalase increase, a NRF2- and FOXO3-mediated antioxidative response (HMOX1 of the endoplasmic reticulum and mitochondrial SOD2) and a NF-?B activation were noted. TGF?1 and proinflammatory cytokines like IL1, IL6 and TNF? were upregulated and stress-related signaling pathways were induced. Sertoli cell mRNA-microarray analysis revealed an increase of TNFR2-signaling components. 53BP1 recruitment and expression levels for DNA repair genes as well as for p53 were elevated and the ones for related sirtuin deacetylases affected (SIRT 1, 3-7) in Sertoli cells. Under chronic Cre-mediated DNA damage conditions a strong downregulation of Sirt1 was observed, suggesting that the decrease of this important coordinator between DNA repair and metabolic signaling might induce the repression release of major transcription factors regulating metabolic and cytokine-mediated stress pathways. Indeed, caspase-3 was activated and increased germ cell apoptosis was observed, suggesting paracrine effects. In conclusion, the observed wide stress-induced effects and metabolic alterations suggest that it is essential to use the correct control animals (Cre/Wt) with matched Cre expression levels to differentiate between Cre-mediated and specific gene-knock out-mediated effects.

KW - Animals

KW - Male

KW - Genotype

KW - Cells, Cultured

KW - Mice

KW - Mice, Inbred C57BL

KW - Mice, Transgenic

KW - Polymerase Chain Reaction

KW - Real-Time Polymerase Chain Reaction

KW - Promoter Regions, Genetic/genetics

KW - Anti-Mullerian Hormone/genetics

KW - Antioxidants/metabolism

KW - Chromosome Walking

KW - Integrases/genetics/metabolism

KW - Peroxisomes/metabolism

KW - Sertoli Cells/metabolism

KW - Signal Transduction/genetics/physiology

KW - Sirtuins/genetics/metabolism

KW - Testis/metabolism

KW - Animals

KW - Male

KW - Genotype

KW - Cells, Cultured

KW - Mice

KW - Mice, Inbred C57BL

KW - Mice, Transgenic

KW - Polymerase Chain Reaction

KW - Real-Time Polymerase Chain Reaction

KW - Promoter Regions, Genetic/genetics

KW - Anti-Mullerian Hormone/genetics

KW - Antioxidants/metabolism

KW - Chromosome Walking

KW - Integrases/genetics/metabolism

KW - Peroxisomes/metabolism

KW - Sertoli Cells/metabolism

KW - Signal Transduction/genetics/physiology

KW - Sirtuins/genetics/metabolism

KW - Testis/metabolism

M3 - SCORING: Journal article

VL - 7

SP - 41097

JO - PLOS ONE

JF - PLOS ONE

SN - 1932-6203

IS - 7

M1 - 7

ER -