title: GATA4 is a key regulator of steroidogenesis and glycolysis in mouse Leydig cells.
abstract: Transcription factor GATA4 is expressed in somatic cells of the mammalian testis. Gene targeting studies in mice have shown that GATA4 is essential for proper differentiation and function of Sertoli cells. The role of GATA4 in Leydig cell development, however, remains controversial, because targeted mutagenesis experiments in mice have not shown a consistent phenotype, possibly due to context-dependent effects or compensatory responses. We therefore undertook a reductionist approach to study the function of GATA4 in Leydig cells. Using microarray analysis and quantitative RT-PCR, we identified a set of genes that are down-regulated or up-regulated after small interfering RNA (siRNA)-mediated silencing of Gata4 in the murine Leydig tumor cell line mLTC-1. These same genes were dysregulated when primary cultures of Gata4(flox/flox) adult Leydig cells were subjected to adenovirus-mediated cre-lox recombination in vitro. Among the down-regulated genes were enzymes of the androgen biosynthetic pathway (Cyp11a1, Hsd3b1, Cyp17a1, and Srd5a). Silencing of Gata4 expression in mLTC-1 cells was accompanied by reduced production of sex steroid precursors, as documented by mass spectrometric analysis. Comprehensive metabolomic analysis of GATA4-deficient mLTC-1 cells showed alteration of other metabolic pathways, notably glycolysis. GATA4-depleted mLTC-1 cells had reduced expression of glycolytic genes (Hk1, Gpi1, Pfkp, and Pgam1), lower intracellular levels of ATP, and increased extracellular levels of glucose. Our findings suggest that GATA4 plays a pivotal role in Leydig cell function and provide novel insights into metabolic regulation in this cell type.
authors: Children's Hospital (A.S., A.K., O.A., M.P., T.-P.A., M.H.), University of Helsinki, Helsinki 00014, Finland; Institute of Biomedicine (O.A.), University of Helsinki, Helsinki 00014, Finland; School of Pharmacy (M.H.), University of Eastern Finland, Kuopio 70211, Finland; Institute of Applied Biotechnology (S.F.), University of Applied Sciences Biberach, Biberach 88400, Germany; Metabolomics Unit (V.V.), Institute for Molecular Medicine Finland, University of Helsinki 00014, Helsinki, Finland; Departments of Physiology and Pediatrics (J.T.), University of Turku, Turku 20520, Finland; and Departments of Pediatrics (A.S., M.P., D.B.W., M.H.) and Developmental Biology (D.B.W.), Washington University in St. Louis, St. Louis, Missouri 63110.
authors: Anja
authors: A
authors: Schrade
authors: Antti
authors: A
authors: Kyrönlahti
authors: Oyediran
authors: O
authors: Akinrinade
authors: Marjut
authors: M
authors: Pihlajoki
authors: Merja
authors: M
authors: Häkkinen
authors: Simon
authors: S
authors: Fischer
authors: Tero-Pekka
authors: TP
authors: Alastalo
authors: Vidya
authors: V
authors: Velagapudi
authors: Jorma
authors: J
authors: Toppari
authors: David B
authors: DB
authors: Wilson
authors: Markku
authors: M
authors: Heikinheimo
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics / Q000378:metabolism; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000728:Androgens / Q000096:biosynthesis*; D000818:Animals; D045744:Cell Line, Tumor; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D050991:GATA4 Transcription Factor / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism; D006019:Glycolysis / Q000235:genetics*; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D025221:Phosphofructokinases / Q000235:genetics / Q000378:metabolism; D010736:Phosphoglycerate Mutase / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D000728:Androgens; D016207:Cytokines; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D015096:3-Hydroxysteroid Dehydrogenases; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase; D025221:Phosphofructokinases; C515658:HK1 protein, mouse; D006593:Hexokinase; D005956:Glucose-6-Phosphate Isomerase; C554960:Gpi1 protein, mouse; D010736:Phosphoglycerate Mutase