PMID- 26698679 OWN - NLM STAT- MEDLINE DCOM- 20160719 LR - 20211203 IS - 1556-5653 (Electronic) IS - 0015-0282 (Print) IS - 0015-0282 (Linking) VI - 105 IP - 3 DP - 2016 Mar TI - Mammalian target of rapamycin controls glucose consumption and redox balance in human Sertoli cells. PG - 825-833.e3 LID - S0015-0282(15)02116-0 [pii] LID - 10.1016/j.fertnstert.2015.11.032 [doi] AB - OBJECTIVE: To study the role of mammalian target of rapamycin (mTOR) in the regulation of human Sertoli cell (hSC) metabolism, mitochondrial activity, and oxidative stress. DESIGN: Experimental study. SETTING: University research center and private assisted reproductive technology centers. PATIENT(S): Six men with anejaculation (psychological, vascular, neurologic) and conserved spermatogenesis. INTERVENTION(S): Testicular biopsies were used from patients under treatment for recovery of male gametes. Primary hSCs cultures were established from each biopsy and divided into a control group and one treated with rapamycin, the inhibitor of mTOR, for 24 hours. MAIN OUTCOME MEASURE(S): Cytotoxicity of hSCs to rapamycin was evaluated by sulforhodamine B assay. The glycolytic profile of hSCs was assessed by proton nuclear magnetic resonance and by studying protein expression of key glycolysis-related transporters and enzymes. Expression of mitochondrial complexes and citrate synthase activity were determined. Protein carbonylation, nitration, lipid peroxidation, and sulfhydryl protein group contents were quantified. The mTOR signaling pathway was studied. RESULT(S): Rapamycin increased glucose consumption by hSCs, maintaining lactate production. Alanine production by rapamycin-exposed hSCs was affected, resulting in an unbalanced intracellular redox state. Rapamycin-exposed hSCs had decreased expression of mitochondrial complex III and increased lipid peroxidation, whereas other oxidative stress markers were unaltered. Treatment of hSCs with rapamycin down-regulated phospho-mTOR (Ser-2448) levels, illustrating an effective partial inhibition of mTORC1. Protein levels of downstream signaling molecule p-4E-BP1 were not altered, suggesting that during treatment it became rephosphorylated. CONCLUSION(S): We show that mTOR regulates the nutritional support of spermatogenesis by hSCs and redox balance in these cells. CI - Copyright (c) 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved. FAU - Jesus, Tito T AU - Jesus TT AD - Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal; CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilha. FAU - Oliveira, Pedro F AU - Oliveira PF AD - Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal; Institute of Health Research an Innovation, Portugal. FAU - Silva, Joaquina AU - Silva J AD - Centre for Reproductive Genetics Prof. Alberto Barros, Porto, Portugal. FAU - Barros, Alberto AU - Barros A AD - Institute of Health Research an Innovation, Portugal; Centre for Reproductive Genetics Prof. Alberto Barros, Porto, Portugal; Department of Genetics, Faculty of Medicine, University of Porto, Porto, Portugal. FAU - Ferreira, Rita AU - Ferreira R AD - Organic Chemistry, Natural and Agrofood Products Centre, Department of Chemistry, University of Aveiro, Aveiro, Portugal. FAU - Sousa, Mario AU - Sousa M AD - Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal; Centre for Reproductive Genetics Prof. Alberto Barros, Porto, Portugal. FAU - Cheng, C Yan AU - Cheng CY AD - Center for Biomedical Research, Population Council, New York, New York. FAU - Silva, Branca M AU - Silva BM AD - CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilha. FAU - Alves, Marco G AU - Alves MG AD - CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilha. Electronic address: alvesmarc@gmail.com. LA - eng GR - R01 HD056034/HD/NICHD NIH HHS/United States GR - U01 HD045908/HD/NICHD NIH HHS/United States GR - U54 HD029990/HD/NICHD NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151214 PL - United States TA - Fertil Steril JT - Fertility and sterility JID - 0372772 RN - 0 (Protein Kinase Inhibitors) RN - EC 2.7.1.1 (MTOR protein, human) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - IY9XDZ35W2 (Glucose) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - Cells, Cultured MH - *Energy Metabolism/drug effects MH - Glucose/*metabolism MH - Glycolysis MH - Humans MH - Lipid Peroxidation MH - Male MH - Mitochondria/enzymology MH - Oxidation-Reduction MH - Oxidative Stress MH - Phosphorylation MH - Primary Cell Culture MH - Protein Carbonylation MH - Protein Kinase Inhibitors/pharmacology MH - Proton Magnetic Resonance Spectroscopy MH - Sertoli Cells/*drug effects/*enzymology MH - Signal Transduction MH - Sirolimus/pharmacology MH - *Spermatogenesis MH - TOR Serine-Threonine Kinases/antagonists & inhibitors/*metabolism PMC - PMC4845725 MID - NIHMS773301 OTO - NOTNLM OT - Sertoli cells OT - mTOR OT - rapamycin OT - spermatogenesis OT - testis EDAT- 2015/12/25 06:00 MHDA- 2016/07/20 06:00 PMCR- 2017/03/01 CRDT- 2015/12/25 06:00 PHST- 2015/06/30 00:00 [received] PHST- 2015/10/16 00:00 [revised] PHST- 2015/11/18 00:00 [accepted] PHST- 2017/03/01 00:00 [pmc-release] PHST- 2015/12/25 06:00 [entrez] PHST- 2015/12/25 06:00 [pubmed] PHST- 2016/07/20 06:00 [medline] AID - S0015-0282(15)02116-0 [pii] AID - 10.1016/j.fertnstert.2015.11.032 [doi] PST - ppublish SO - Fertil Steril. 2016 Mar;105(3):825-833.e3. doi: 10.1016/j.fertnstert.2015.11.032. Epub 2015 Dec 14.