GATA4 regulates blood-testis barrier function and lactate metabolism in mouse sertoli cells

Anja Schrade, Antti Kyrönlahti, Oyediran Akinrinade, Marjut Pihlajoki, Simon Fischer, Verena Martinez Rodriguez, Kerstin Otte, Vidya Velagapudi, Jorma Toppari, David B. Wilson, Markku Heikinheimo

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Conditional deletion of Gata4 in Sertoli cells (SCs) of adult mice has been shown to increase permeabilityofthe blood-testis barrier (BTB) and disrupt spermatogenesis.Togain insight into the molecular underpinnings of these phenotypic abnormalities, we assessed the impact of Gata4gene silencing in cell culture models. Microarray hybridization identified genes dysregulated by siRNA-mediated inhibition of Gata4 in TM4 cells, an immortalized mouse SC line. Differentially expressed genes were validated by quantitative RT-PCRanalysisofprimarycultures of Gata4 flox/flox mouse SCs that had been subjected to cre-mediated recombination in vitro. Depletion of GATA4 in TM4 cells and primary SCs was associated with altered expression of genes involved in key facets of BTB maintenance, including tight/adherens junction formation (Tjp1, Cldn12, Vcl, Tnc, Csk) and extracellular matrix reorganization (Lamc1, Col4a1, Col4a5, Mmp10, Mmp23, Timp2). Western blotting and immunocytochemistry demonstrated reduced levelsof tight junction protein-1,aprototypical tight junction protein, in GATA4-depleted cells. These changes were accompanied by a loss of morphologically recognizable junctional complexes and a decline in epithelial membrane resistance. Furthermore, Gata4 gene silencing was associated with altered expression of Hk1, Gpi1, Pfkp, Pgam1, Gls2, Pdk3, Pkd4, and Ldhb, genes regulating the production of lactate,a key nutrient that SCs provide to developing germ cells. Comprehensive metabolomic profiling demonstrated impaired lactate production in GATA4-deficient SCs. We conclude that GATA4 plays a pivotal role in the regulation of BTB function and lactate metabolism in mouse SCs.

Original languageEnglish
Pages (from-to)2416-2431
Number of pages16
JournalEndocrinology
Volume157
Issue number6
DOIs
StatePublished - Jun 2016

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